Designed escrt-recruiting domain (ERD) adapter systems and co-localized immune cell-targeting proteins induce efficient budding of enveloped nanoparticles (ENPS) that display high levels of immunogens

Abstract

Disclosed herein include methods, compositions, and kits suitable for use, e.g., in vaccination. Provided are nucleic acid compositions (e.g., mRNA vaccines, DNA vaccines) comprising a polynucleotide encoding at least one of a dimerization fusion protein, an adapter fusion protein, a recombinant protein, and a chimeric protein. The recombinant protein or the dimerization fusion protein can comprise an optional antigenic polypeptide (AP) and the adapter fusion protein, the recombinant protein or the chimeric protein can comprise an endosomal sorting complex required for transport (ESCRT)-recruiting domain (ERD). Recruitment of one or more ESCRT proteins results in secretion of enveloped nanoparticles (ENPs) from a cell in which at least one of a dimerization fusion protein, an adapter fusion protein, a recombinant protein, and a chimeric protein are expressed. There are also provided populations of ENPs in some embodiments.

Description

DESIGNED ESCRT-RECRUITING DOMAIN (ERD) ADAPTER SYSTEMS AND COLOCALIZED IMMUNE CELL-TARGETING PROTEINS INDUCE EFFICIENT BUDDING OF ENVELOPED NANOPARTICLES (ENPS) THAT DISPLAY HIGH LEVELS OF IMMUNOGENSRELATED APPLICATIONS

[0001] The present application claims priority to U.S. Provisional Application No. 63 / 654,377, filed May 31, 2024. The entire content of this application is hereby expressly incorporated by reference in its entirety.STATEMENT REGARDING FEDERALLY SPONSORED R&D

[0002] This invention was made with government support under Grant No. OD033362 awarded by the National Institutes of Health. The government has certain rights in the invention.REFERENCE TO SEQUENCE LISTING

[0003] The present application is being filed along with a Sequence Listing in electronic format. The Sequence Listing is provided as a file entitled 30KJ-365872- WO_SequenceListing, created May 22, 2025, which is 152 kilobytes in size. The information in the electronic format of the Sequence Listing is incorporated herein by reference in its entirety.BACKGROUNDField

[0004] The present disclosure relates generally to the field of vaccines.Description of the Related Art

[0005] mRNA vaccines have emerged as an excellent platform for vaccine development. Hybrid mRNA vaccine approaches utilize genetically encoded self-assembling enveloped nanoparticles (ENPs) to combine features of mRNA- and nanoparticle-based vaccines. ENP self-assembly is achieved by inserting an ESCRT -recruiting domain (ERD) into the cytoplasmic domain of the membrane-bound immunogen, which recruits host cell proteins from the endosomal sorting complex required for transport (ESCRT) pathway. Mouse studies showed that immunizations with mRNA encoding a SARS-CoV-2 spike-ERD construct elicited markedly higher binding and neutralizing antibody titers against original and variant SARS-CoV-2 compared to conventional spike mRNA and protein-based spike nanoparticles.

[0006] There is a need for improved ERD constructs that promote high levels of both cell surface expression and ENP budding for more optimal stimulation of the immune system.SUMMARY

[0007] Disclosed herein include compositions. In some embodiments, the composition comprises: a nucleic acid composition comprising: (i) a polynucleotide encoding a dimerization fusion protein, wherein the dimerization fusion protein comprises an optional antigenic polypeptide (AP) and a heterologous cytoplasmic tail, and (ii) a polynucleotide encoding an adapter fusion protein comprising an endosomal sorting complex required for transport (ESCRT)- recruiting domain (ERD) and an adapter domain capable of binding the heterologous cytoplasmic tail to form a heterodimer, wherein binding of the adapter domain to the heterologous cytoplasmic tail is capable of recruiting one or more ESCRT proteins to the heterodimer, thereby inducing a plurality of dimerization fusion proteins to self-assemble into an enveloped nanoparticle (ENP) secreted from a cell in which the dimerization fusion protein and adapter fusion protein are expressed, thereby generating a population of ENPs.

[0008] In some embodiments, the composition comprises: a nucleic acid composition comprising (i) n polynucleotides each encoding an nth dimerization fusion protein, wherein n is an integer from 2 to 500, wherein each dimerization fusion protein comprises an optional antigenic polypeptide (AP) and a heterologous cytoplasmic tail, wherein at least two of the dimerization fusion proteins differ with respect to the AP; and (ii) a polynucleotide encoding an adapter fusion protein comprising an endosomal sorting complex required for transport (ESCRT)-recruiting domain (ERD) and an adapter domain capable of binding the heterologous cytoplasmic tail to form a heterodimer, wherein binding of the adapter domain to the heterologous cytoplasmic tail is capable of recruiting one or more ESCRT proteins to the heterodimer, thereby inducing a plurality of dimerization fusion proteins to self-assemble into an enveloped nanoparticle (ENP) secreted from a cell in which the n dimerization fusion proteins and the adapter fusion protein are expressed, thereby generating a population of ENPs.

[0009] In some embodiments, the dimerization fusion proteins are capable of being presented on the surface of a cell in which the dimerization fusion proteins are expressed. In some embodiments, the self-assembly of an ENP: does not require an exogenous nucleic acid other than the nucleic acid composition, and / or does not require any exogenous components other than the dimerization fusion proteins and the adapter fusion proteins.

[0010] In some embodiments, the cell is: a cell of a subject; an in vivo cell, an ex vivo cell, or an in situ cell; and / or an adherent cell or a suspension cell. In some embodiments, upon secretion from a cell of a subject, the ENPs are capable of distributing within one or more tissues of the subject. In some embodiments, the one or more tissues comprise adrenal gland tissue, appendix tissue, bladder tissue, bone, bowel tissue, brain tissue, breast tissue, bronchi, coronal tissue, ear tissue, esophagus tissue, eye tissue, gall bladder tissue, genital tissue, heart tissue,hypothalamus tissue, kidney tissue, large intestine tissue, intestinal tissue, larynx tissue, liver tissue, lung tissue, lymph nodes, mouth tissue, nose tissue, pancreatic tissue, parathyroid gland tissue, pituitary gland tissue, prostate tissue, rectal tissue, salivary gland tissue, skeletal muscle tissue, skin tissue, small intestine tissue, spinal cord, spleen tissue, stomach tissue, thymus gland tissue, trachea tissue, thyroid tissue, ureter tissue, urethra tissue, soft and connective tissue, peritoneal tissue, blood vessel tissue, fat tissue, or any combination thereof. In some embodiments, the ENPs engage a plurality of immune cells in the one or more tissues, thereby mimicking a natural infection.

[0011] Disclosed herein include compositions. In some embodiments, the composition comprises: a population of enveloped nanoparticles (ENPs), wherein each of the ENPs comprises a plurality of the dimerization fusion proteins each comprising a heterologous cytoplasmic tail and optionally, an antigenic polypeptide (AP).

[0012] In some embodiments, the adapter fusion protein comprises, from N-terminus to C-terminus: the adapter domain, an optional linker, and the ERD. In some embodiments, the linker: is a flexible linker, a rigid linker, or a hybrid linker; is hydrophilic or hydrophobic; is between 1 and 250 amino acids; comprises one or more flexible amino acid residues. In some embodiments, the linker comprises about 1 to about 250 flexible amino acid residues. In some embodiments, the flexible amino acid residues comprise glycine, serine, or a combination thereof. In some embodiments, the linker comprises 3 repeating amino acid subunits or more.

[0013] In some embodiments, the heterologous cytoplasmic tail is derived from or comprises a cytoplasmic tail of a cell surface protein, and the adapter domain is (1) capable of binding the heterologous cytoplasmic tail derived from or comprising the cytoplasmic tail of said cell surface protein and (2) capable of targeting the adapter fusion protein to the plasma membrane. In some embodiments, the cell surface protein is or is derived from a human protein, a non-human mammalian protein, an avian protein, or a reptile protein and / or the adapter domain is or is derived from a human protein, a non-human mammalian protein, an avian protein, or a reptile protein.

[0014] In some embodiments, the heterologous cytoplasmic tail comprises or is derived from a cytoplasmic tail (CT) of CD4 or CD8; and / or the adapter domain comprises or is derived from at least a portion of Lek tyrosine kinase. In some embodiments, the CD4 CT comprises the sequence of SEQ ID NO: 95 or SEQ ID NO: 96. In some embodiments, the CD8 CT comprises the sequence of SEQ ID NO: 101. In some embodiments, the adapter domain comprises the sequence of SEQ ID NO: 43 or SEQ ID NO: 97. In some embodiments, the heterologous cytoplasmic tail of CD4 comprises the sequence of SEQ ID NO: 98, SEQ ID NO: 99, or SEQ ID NO: 100. In some embodiments: the heterologous cytoplasmic tail of CD4comprises the sequence of SEQ ID NO: 102, and the adapter domain comprises the sequence of SEQ ID NO: 105; the heterologous cytoplasmic tail of CD4 comprises the sequence of SEQ ID NO: 103, and the adapter domain comprises the sequence of SEQ ID NO: 106; or the heterologous cytoplasmic tail of CD4 comprises the sequence of SEQ ID NO: 104, and the adapter domain comprises the sequence of SEQ ID NO: 107.

[0015] In some embodiments, the composition further comprises: a polynucleotide encoding a second adapter fusion protein comprising an endosomal sorting complex required for transport (ESCRT)-recruiting domain (ERD) and an adapter domain capable of binding the heterologous cytoplasmic tail to form a heterodimer, wherein the adapter domain of the second adapter fusion protein is the same or different from the adapter domain of the adapter fusion protein. In some embodiments, the ERD of the second adapter fusion protein is the same or different from the ERD of the adapter fusion protein. In some embodiments, the adapter domain of the second adapter fusion protein comprises the sequence of SEQ ID NO: 109 and the adapter domain of the adapter fusion protein comprises the sequence of SEQ ID NO: 43 or SEQ ID NO: 97.

[0016] In some embodiments, the heterologous cytoplasmic tail comprises or is derived from G. gallus CD4 CT. In some embodiments, the heterologous cytoplasmic tail comprises the sequence of SEQ ID NO: 110. In some embodiments, the adapter domain comprises or is derived from at least a portion of G. gallus Lek tyrosine kinase. In some embodiments, the adapter domain comprises the sequence of SEQ ID NO: 113. In some embodiments, the heterologous cytoplasmic tail comprises or is derived from P. vitticeps CD4 CT. In some embodiments, the heterologous cytoplasmic tail comprises the sequence of SEQ ID NO: 111. In some embodiments, the adapter domain comprises or is derived from at least a portion of P. vitticeps Lek tyrosine kinase. In some embodiments, the adapter domain comprises the sequence of SEQ ID NO: 114. In some embodiments, the heterologous cytoplasmic tail comprises or is derived from N. scutatus CD4 CT. In some embodiments, the heterologous cytoplasmic tail comprises the sequence of SEQ ID NO: 112. In some embodiments, the adapter domain comprises or is derived from at least a portion of N. scutatus Lek tyrosine kinase. In some embodiments, the adapter domain comprises the sequence of SEQ ID NO: 115.

[0017] In some embodiments, the dimerization fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 41-42, 45, 48-59, 63, 68-69, and 83-85. In some embodiments, the adapter fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 44, 46, 60-62, 64-65, 70-82, and 86-88. In some embodiments, (i) the dimerization fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 51 ; and (ii) the adapter fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 44, 64, 77, and 80-82. In some embodiments, (i) the dimerization fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 51 ; and (ii) the adapter fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 44. In some embodiments, (i) the dimerization fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 51 ; and (ii) the adapter fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 64. In some embodiments, (i) the dimerization fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 51; and (ii) the adapter fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 77. In some embodiments, (i) the dimerization fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 51 ; and (ii) the adapter fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 80. In some embodiments, (i) the dimerization fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 51; and (ii) the adapter fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 81. In some embodiments, (i) the dimerization fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 51; and (ii) the adapter fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 82.

[0018] In some embodiments, (i) the dimerization fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 69; and (ii) the adapter fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequenceidentity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 44, 64, 77, and 80-82. In some embodiments, (i) the dimerization fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 69; and (ii) the adapter fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 44. In some embodiments, (i) the dimerization fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 69; and (ii) the adapter fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 64. In some embodiments, (i) the dimerization fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 69; and (ii) the adapter fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 77. In some embodiments, (i) the dimerization fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 69; and (ii) the adapter fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 80. In some embodiments, (i) the dimerization fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 69; and (ii) the adapter fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 81. In some embodiments, (i) the dimerization fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 69; and (ii) the adapter fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 82.

[0019] In some embodiments, the dimerization fusion protein comprises, from N- terminus to C-terminus: the antigenic polypeptide, an optional linker, and the heterologous cytoplasmic tail. In some embodiments the dimerization fusion protein comprises an endogenous cytoplasmic tail N-terminal to the heterologous cytoplasmic tail. In some embodiments, the dimerization fusion protein comprises from C-terminus to N-terminus: the antigenic polypeptide, an optional linker, and the heterologous cytoplasmic tail, optionally wherein the dimerization fusion protein comprises an endogenous cytoplasmic tail C-terminal to the heterologous cytoplasmic tail. In some embodiments, the adapter fusion protein comprises, from N-terminus toC-terminus: a heterologous membrane-targeting domain, an optional first linker, the adapter domain, an optional second linker, and the ERD. In some embodiments, the linker, the first linker, and / or the second linker: is a flexible linker, a rigid linker, or a hybrid linker; is hydrophilic or hydrophobic; is between 1 and 250 amino acids; comprises one or more flexible amino acid residues, optionally about 1 to about 250 flexible amino acid residues, further optionally the flexible amino acid residues comprise glycine, serine, or a combination thereof; and / or comprises 3 repeating amino acid subunits or more.

[0020] In some embodiments, the heterologous membrane targeting domain comprises or is derived from at least a portion of LAT, PAG, LCK, FYN, LAX, CD2, CD3, CD4, CD5, CD7, CD8a, PD1, SRC, or LYN. In some embodiments, the heterologous membrane targeting domain comprises or is derived from FYN. In some embodiments, the heterologous membrane targeting domain comprises the sequence of SEQ ID NO: 118.

[0021] In some embodiments, the heterologous cytoplasmic tail and / or the adapter domain are each selected from the group comprising DHD9 heterodimer a, DHD13 XAAA heterodimer a, DHD13 XAXA heterodimer a, DHD13 XAAX heterodimer a, DHD13_2:341 heterodimer a, DHD13 AAAA heterodimer a, DHD13 BAAA heterodimer a, DHD13_4: 123 heterodimer a, DHD13_1 :234 heterodimer a, DHD15 heterodimer a, DHD20 heterodimer a, DHD21 heterodimer a, DHD25 heterodimer a, DHD27 heterodimer a, DHD30 heterodimer a, DHD33 heterodimer a, DHD34 XAAXA heterodimer a, DHD34 XAXXA heterodimer a, DHD34 XAAAA heterodimer a, DHD36 heterodimer a, DHD37 ABXB heterodimer a, DHD37 BBBB heterodimer a, DHD37 XBXB heterodimer a, DHD37 AXXB heterodimer a, DHD37_3: 124 heterodimer a, DHD37_1 :234 heterodimer a, DHD37 AXBB heterodimer a, DHD37 XBBA heterodimer a, DHD39 heterodimer a, DHD40 heterodimer a, DHD43 heterodimer a, DHD65 heterodimer a, DHD70 heterodimer a, DHD88 heterodimer a, DHD89 heterodimer a, DHD90 heterodimer a, DHD91 heterodimer a, DHD92 heterodimer a, DHD93 heterodimer a, DHD94 heterodimer a, DHD94_3:214 heterodimer a, DHD94_2: 143 heterodimer a, DHD95 heterodimer a, DHD96 heterodimer a, DHD97 heterodimer a, DHD98 heterodimer a, DHD99 heterodimer a, DHD100 heterodimer a, DHD101 heterodimer a, DHD102 heterodimer a, DHD102_l :243 heterodimer a, DHD103 heterodimer a, DHD103_l :423 heterodimer a, DHD104 heterodimer a, DHD105 heterodimer a, DHD106 heterodimer a, DHD107 heterodimer a, DHD108 heterodimer a, DHD109 heterodimer a, DHD110 heterodimer a, DHD111 heterodimer a, DHD112 heterodimer a, DHD113 heterodimer a, DHD114 heterodimer a, DHD115 heterodimer a, DHD116 heterodimer a, DHD117 heterodimer a, DHD118 heterodimer a, DHD119 heterodimer a, DHD120 heterodimer a, DHD121 heterodimer a, DHD122 heterodimer a, DHD123 heterodimer a, DHD124 heterodimer a, DHD125 heterodimer a, DHD126heterodimer a, DHD127 heterodimer a, DHD128 heterodimer a, DHD129 heterodimer a, DHD130 heterodimer a, DHD145 heterodimer a, DHD146 heterodimer a, DHD147 heterodimer a, DHD1 heterodimer a, DHD2 heterodimer a, DHD3 heterodimer a, DHD4 heterodimer a, DHD5 heterodimer a, DHD6 heterodimer a, DHD7 heterodimer a, DHD8 heterodimer a, DHD16 heterodimer a, DHD18 heterodimer a, DHD19 heterodimer a, DHD22 heterodimer a, DHD23 heterodimer a, DHD24 heterodimer a, DHD26 heterodimer a, DHD28 heterodimer a, DHD29 heterodimer a, DHD31 heterodimer a, DHD32 heterodimer a, DHD38 heterodimer a, DHD60 heterodimer a, DHD63 heterodimer a, DHD66 heterodimer a, DHD67 heterodimer a, DHD69 heterodimer a, DHD71 heterodimer a, DHD72 heterodimer a, DHD73 heterodimer a, DHD148 heterodimer a, DHD149 heterodimer a, DHD150 heterodimer a, DHD151 heterodimer a, DHD152 heterodimer a, DHD153 heterodimer a, DHD154 heterodimer a, DHD155 heterodimer a, DHD156 heterodimer a, DHD157 heterodimer a, DHD158 heterodimer a, DHD159 heterodimer, DH160 heterodimer a, DHD161 heterodimer a, DHD162 heterodimer a, DHD163 heterodimer a, DHD164 heterodimer a, DHD165 heterodimer a, DHD166 heterodimer a, DHS17 heterodimer a, DHD17 heterodimer a, DHD131 heterodimer a, DHD132 heterodimer a, DHD133 heterodimer a, DHD134 heterodimer a, DHD135 heterodimer a, DHD136 heterodimer, DH137 heterodimer a, DHD138 heterodimer a, DHD139 heterodimer a, DHD140 heterodimer a, DHD141 heterodimer a, DHD142 heterodimer a, DHD143 heterodimer a, DHD144 heterodimer a, DHD9 heterodimer b, DHD13 XAAA heterodimer b, DHD13 XAXA heterodimer b, DHD13 XAAX heterodimer b, DHD13_2:341 heterodimer b, DHD13 AAAA heterodimer b, DHD13 BAAA heterodimer b, DHD13_4: 123 heterodimer b, DHD13_1 :234 heterodimer b, DHD15 heterodimer b, DHD20 heterodimer b, DHD21 heterodimer b, DHD25 heterodimer b, DHD27 heterodimer b, DHD30 heterodimer b, DHD33 heterodimer b, DHD34 XAAXA heterodimer b, DHD34 XAXXA heterodimer b, DHD34 XAAAA heterodimer b, DHD36 heterodimer b, DHD37 ABXB heterodimer b, DHD37 BBBB heterodimer b, DHD37_XBXB heterodimer b, DHD37 AXXB heterodimer b, DHD37_3: 124 heterodimer b, DHD37_1 :234 heterodimer b, DHD37 AXBB heterodimer b, DHD37 XBBA heterodimer b, DHD39 heterodimer b, DHD40 heterodimer b, DHD43 heterodimer b, DHD65 heterodimer b, DHD70 heterodimer b, DHD88 heterodimer b, DHD89 heterodimer b, DHD90 heterodimer b, DHD91 heterodimer b, DHD92 heterodimer b, DHD93 heterodimer b, DHD94 heterodimer b, DHD94_3 :214 heterodimer b, DHD94_2: 143 heterodimer b, DHD95 heterodimer b, DHD96 heterodimer b, DHD97 heterodimer b, DHD98 heterodimer b, DHD99 heterodimer b, DHD100 heterodimer b, DHD101 heterodimer b, DHD102 heterodimer b, DHD102_l :243 heterodimer b, DHD103 heterodimer b, DHD103_l :423 heterodimer b, DHD104 heterodimer b, DHD105 heterodimer b, DHD106 heterodimer b, DHD107 heterodimer b, DHD108 heterodimer b,DHD109 heterodimer b, DHD110 heterodimer b, DHD111 heterodimer b, DHD112 heterodimer b, DHD113 heterodimer b, DHD114 heterodimer b, DHD115 heterodimer b, DHD116 heterodimer b, DHD117 heterodimer b, DHD118 heterodimer b, DHD119 heterodimer b, DHD120 heterodimer b, DHD121 heterodimer b, DHD122 heterodimer b, DHD123 heterodimer b, DHD124 heterodimer b, DHD125 heterodimer b, DHD126 heterodimer b, DHD127 heterodimer b, DHD128 heterodimer b, DHD129 heterodimer b, DHD130 heterodimer b, DHD145 heterodimer b, DHD146 heterodimer b, DHD147 heterodimer b, DHD1 heterodimer b, DHD2 heterodimer b, DHD3 heterodimer b, DHD4 heterodimer b, DHD5 heterodimer b, DHD6 heterodimer b, DHD7 heterodimer b, DHD8 heterodimer b, DHD16 heterodimer b, DHD18 heterodimer b, DHD19 heterodimer b, DHD22 heterodimer b, DHD23 heterodimer b, DHD24 heterodimer b, DHD26 heterodimer b, DHD28 heterodimer b, DHD29 heterodimer b, DHD31 heterodimer b, DHD32 heterodimer b, DHD38 heterodimer b, DHD60 heterodimer b, DHD63 heterodimer b, DHD66 heterodimer b, DHD67 heterodimer b, DHD69 heterodimer b, DHD71 heterodimer b, DHD72 heterodimer b, DHD73 heterodimer b, DHD148 heterodimer b, DHD149 heterodimer b, DHD150 heterodimer b, DHD151 heterodimer b, DHD152 heterodimer b, DHD153 heterodimer b, DHD154 heterodimer b, DHD155 heterodimer b, DHD156 heterodimer b, DHD157 heterodimer b, DHD158 heterodimer b, DHD159 heterodimer b, DHD160 heterodimer b, DHD161 heterodimer b, DHD162 heterodimer b, DHD163 heterodimer b, DHD164 heterodimer b, DHD165 heterodimer b, DHD166 heterodimer b, DHS17 heterodimer b, DHD17 heterodimer b, DHD131 heterodimer b, DHD132 heterodimer b, DHD133 heterodimer b, DHD134 heterodimer b, DHD135 heterodimer b, DHD136 heterodimer b, DHD137 heterodimer b, DHD138 heterodimer b, DHD139 heterodimer b, DHD140 heterodimer b, DHD141 heterodimer b, DHD142 heterodimer b, DHD143 heterodimer b, DHD144 heterodimer b, portions thereof, derivatives thereof, or any combination thereof.

[0022] In some embodiments, the heterologous cytoplasmic tail and / or the adapter domain comprises or is derived from SYNZIP1, SYNZIP2, SYNZIP3, SYNZIP4, SYNZIP5, SYNZIP6, SYNZIP7, SYNZIP8, SYNZIP9, SYNZIP10, SYNZIP11, SYNZIP12, SYNZIP13, SYNZIP14, SYNZIP15, SYNZIP16, SYNZIP17, SYNZIP18, SYNZIP19, SYNZIP20, SYNZIP21, SYNZIP22, SYNZIP23, BATF, FOS, ATF4, BACH1, JUNE), NFE2L3, Azip, Bzip, a PDZ domain ligand, an SH3 domain, a PDZ domain, a GTPase binding domain, a leucine zipper domain, an SH2 domain, a PTB domain, an FHA domain, a WW domain, a 14-3- 3 domain, a death domain, a caspase recruitment domain, a bromodomain, a chromatin organization modifier, a shadow chromo domain, an F-box domain, a HECT domain, a RING finger domain, a sterile alpha motif domain, a glycine-tyrosine-phenylalanine domain, a SNAP domain, a VHS domain, an ANK repeat, an armadillo repeat, a WD40 repeat, an MH2 domain, acalponin homology domain, a Dbl homology domain, a gelsolin homology domain, a PB 1 domain, a SOCS box, an RGS domain, a Toll / IL-1 receptor domain, a tetratricopeptide repeat, a TRAF domain, a Bcl-2 homology domain, a coiled-coil domain, a bZIP domain, portions thereof, variants thereof, or any combination thereof.

[0023] In some embodiments, the heterologous cytoplasmic tail comprises or is derived from ACIDpl or BASEp 1 or the heterologous cytoplasmic tail comprises or is derived from N5 or N6. In some embodiments, the heterologous cytoplasmic tail comprises the sequence of any one of SEQ ID NOs: 116-117 and 121-122. In some embodiments, the adapter domain comprises or is derived from ACIDpl or BASEp 1 or the heterologous cytoplasmic tail comprises or is derived from N5 or N6. In some embodiments, the adapter domain comprises the sequence of any one of SEQ ID NOs: 116-117 and 121-122. In some embodiments, the dimerization fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 89-90 and 93. In some embodiments, the adapter fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 91-92 and 94. In some embodiments, (i) the dimerization fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 93; and (ii) the adapter fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 94. In some embodiments, (i) the polynucleotide encoding the dimerization fusion protein or the n polynucleotides each encoding an nth dimerization fusion protein, and (ii) the polynucleotide encoding the adapter fusion protein, are each present in a different nucleic acid molecule.

[0024] In some embodiments, the amount of (i) the polynucleotide encoding the dimerization fusion protein or the n polynucleotides each encoding an nth dimerization fusion protein; and (ii) the polynucleotide encoding the adapter fusion protein, are present in the composition at a molar ratio of about 9: 1, 5: 1, or 1 : 1. In some embodiments, (i) the polynucleotide encoding the dimerization fusion protein or the n polynucleotides each encoding an nth dimerization fusion protein, and (ii) the polynucleotide encoding the adapter fusion protein, are present in the same nucleic acid molecule. In some embodiments, the cell expressing the dimerization fusion protein or the plurality of dimerization fusion proteins and the adapter fusion protein exhibits at least a 2-fold increase in AP expression at the surface of the cell, relative to a cell expressing a fusion protein or a plurality of fusion proteins comprising an AP and an ERD and does not express the adapter fusion protein. In some embodiments, the production of ENPsfrom the cell expressing the dimerization fusion protein or the plurality of dimerization fusion proteins and the adapter fusion protein is increased by at least 2-fold, relative to a cell expressing a fusion protein or a plurality of fusion proteins comprising an AP and an ERD and does not express the adapter fusion protein.

[0025] In some embodiments, the nucleic acid composition further comprises a polynucleotide comprising or encoding a tetherin inhibitor. In some embodiments, the tetherin inhibitor is capable of modulating expression, concentration, localization, stability, and / or activity of tetherin. In some embodiments, the tetherin inhibitor comprises a dsRNA, an siRNA, an shRNA, a pre-miRNA, a pri-miRNA, a miRNA, an stRNA, an IncRNA, a piRNA, a snoRNA, or a protein. In some embodiments, one or more of (i) the polynucleotide comprising or encoding the tetherin inhibitor, (ii) the polynucleotide encoding the dimerization fusion protein or the n polynucleotides each encoding an nth dimerization fusion protein, and (iii) the polynucleotide encoding the adapter fusion protein, are present in a same or a different nucleic acid molecule. In some embodiments, the amount of (i) the polynucleotide comprising or encoding the tetherin inhibitor; and (ii) the polynucleotide encoding the dimerization fusion protein or the n polynucleotides each encoding an nth dimerization fusion protein, and / or the polynucleotide encoding the adapter fusion protein, are present in the composition at a molar ratio of about 1 : 1, 1 :5 or 1 :25. In some embodiments, the polynucleotide comprising or encoding the tetherin inhibitor and the polynucleotide encoding the dimerization fusion protein are present in the same nucleic acid. In some embodiments, the polynucleotide comprising or encoding the tetherin inhibitor and the polynucleotide encoding the adapter fusion protein are present in the same nucleic acid. In some embodiments, the tetherin inhibitor comprises or is derived from a viral protein, optionally the virus is HIV-1, HIV-2, SIV, Ebola virus, KSHV, SARS CoV, or SARS- CoV-2. In some embodiments, the tetherin inhibitor comprises HIV-1 Vpu protein, KSHV K5 protein, SARS-CoV-2 ORF7a, HIV-2 Env, Ebola GP, SIV Env, SIV Vpu, SIV Nef, or any portions, variants or derivatives thereof. In some embodiments, the tetherin inhibitor comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 132-134. In some embodiments, the tetherin inhibitor comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 132. In some embodiments, presence or expression of the tetherin inhibitor in the cell results in an increase in ENP production by the cell by at least 2-fold, relative to a cell that does not comprise or express the tetherin inhibitor.

[0026] Disclosed herein include compositions. In some embodiments, the composition comprises: a nucleic acid composition comprising: (i) a polynucleotide encoding a recombinantprotein comprising an optional antigenic polypeptide (AP) and a transmembrane domain, and (ii) a polynucleotide encoding a chimeric protein comprising: a) a signal peptide, b) a cell surface domain, c) a transmembrane domain, and d) a cytoplasmic domain; wherein the recombinant protein and / or the chimeric protein further comprises an endosomal sorting complex required for transport (ESCRT)-recruiting domain (ERD), optionally the recombinant protein further comprises a cytoplasmic domain; and wherein the recombinant protein and the chimeric protein are capable of co-localizing to a site on the plasma membrane of the cell, and wherein the ERD is capable of recruiting one or more ESCRT proteins to the site on the plasma membrane, thereby inducing a plurality of recombinant proteins and chimeric proteins to self-assemble into an enveloped nanoparticle (ENP) secreted from a cell in which the recombinant protein and chimeric protein are expressed, thereby generating a population of ENPs, optionally the cytoplasmic domain of the recombinant protein and the cytoplasmic domain of the chimeric protein are capable of co-localizing the recombinant protein and the chimeric protein to a site on the plasma membrane of the cell, optionally the transmembrane domain of the recombinant protein and the transmembrane domain of the chimeric protein are capable of co-localizing the recombinant protein and the chimeric protein to a site on the plasma membrane of the cell.

[0027] Disclosed herein include compositions. In some embodiments, the composition comprises: a nucleic acid composition comprising: (i) n polynucleotides each encoding a recombinant protein comprising an optional antigenic polypeptide (AP) and a transmembrane domain, wherein at least two of the recombinant proteins differ with respect to the AP, wherein n is an integer from 2 to 500; and (ii) z polynucleotides each encoding a chimeric protein comprising: a) a signal peptide, b) a cell surface domain, wherein at least two of the chimeric proteins differ with respect to the cell surface domain, c) a transmembrane domain, and d) a cytoplasmic domain wherein z is an integer from 1 to 500, wherein the nth recombinant proteins and / or the zth chimeric proteins further comprise an endosomal sorting complex required for transport (ESCRT)-recruiting domain (ERD), optionally the nth recombinant proteins further comprise a cytoplasmic domain, and wherein each of the nth recombinant protein and each of the zth chimeric protein are capable of co-localizing to a site on the plasma membrane of the cell, and the ERD is capable of recruiting one or more ESCRT proteins to the site on the plasma membrane, thereby inducing a plurality of recombinant proteins and chimeric proteins to self-assemble into an enveloped nanoparticle (ENP) secreted from the cell in which the recombinant proteins and chimeric proteins are expressed, thereby generating a population of ENPs, optionally the cytoplasmic domain of each of the nth recombinant protein and the cytoplasmic domain of each of the zth chimeric protein are capable of co-localizing each of the nth recombinant protein and each of the zth chimeric protein to a site on the plasma membrane of the cell, optionally thetransmembrane domain of each of the nth recombinant protein and the transmembrane domain of each of the zth chimeric protein are capable of co-localizing each of the nth recombinant protein and each of the zth chimeric protein to a site on the plasma membrane of the cell.

[0028] In some embodiments, the recombinant proteins and chimeric proteins are capable of being presented on the surface of the cell in which the plurality of recombinant proteins and the plurality of chimeric proteins are expressed. In some embodiments, the self-assembly of an ENP: does not require an exogenous nucleic acid other than the nucleic acid composition, and / or does not require any exogenous components other than the recombinant protein and the chimeric protein.

[0029] In some embodiments, the cell is: a cell of a subject; an in vivo cell, an ex vivo cell, or an in situ cell; and / or an adherent cell or a suspension cell. In some embodiments, upon secretion from a cell of a subject, the ENPs are capable of distributing within one or more tissues of the subject. In some embodiments, the one or more tissues comprise adrenal gland tissue, appendix tissue, bladder tissue, bone, bowel tissue, brain tissue, breast tissue, bronchi, coronal tissue, ear tissue, esophagus tissue, eye tissue, gall bladder tissue, genital tissue, heart tissue, hypothalamus tissue, kidney tissue, large intestine tissue, intestinal tissue, larynx tissue, liver tissue, lung tissue, lymph nodes, mouth tissue, nose tissue, pancreatic tissue, parathyroid gland tissue, pituitary gland tissue, prostate tissue, rectal tissue, salivary gland tissue, skeletal muscle tissue, skin tissue, small intestine tissue, spinal cord, spleen tissue, stomach tissue, thymus gland tissue, trachea tissue, thyroid tissue, ureter tissue, urethra tissue, soft and connective tissue, peritoneal tissue, blood vessel tissue, fat tissue, or any combination thereof. In some embodiments, the ENPs engage a plurality of immune cells in the one or more tissues, thereby mimicking a natural infection.

[0030] Disclosed herein include compositions. In some embodiments, the composition comprises: a population of enveloped nanoparticles (ENPs), wherein each of the ENPs comprises a plurality of recombinant proteins comprising a transmembrane domain and an optional antigenic polypeptide (AP), and a plurality of chimeric proteins comprising: a) a signal peptide, b) a cell surface domain, c) a transmembrane domain, and d) a cytoplasmic domain, wherein the plurality of recombinant proteins and / or the plurality of chimeric proteins further comprise an endosomal sorting complex required fortransport (ESCRT)-recruiting domain (ERD), optionally the plurality of recombinant proteins further comprise a cytoplasmic domain.

[0031] In some embodiments, the ENPs are derived from expression of any of the nucleic acid compositions of the disclosure. In some embodiments, the signal peptide comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to an amino acid of SEQ ID NO: 127 or a sequence having one, two, or threemismatches relative to the sequence of SEQ ID NO: 127.

[0032] In some embodiments, the cell surface domain comprises an immune-cell targeting polypeptide or a second antigenic polypeptide sequence. In some embodiments, the immune-cell targeting polypeptide comprises or is derived from one or more of complement component 3d (C3d), flagellin, an influenza HA molecule, a parainfluenza FiN molecule, a Venezuelan equine encephalitis (VEE) glycoprotein molecule, a mannose receptor molecule, a mammalian toll- like receptor (TLR) ligand molecule, a MIP-1 alpha molecule, a RANTES MIP- 1 beta molecule, a GM-CSF molecule, a Flt3 ligand molecule, a CD40 ligand molecule, a Prevotella intermedia glycoprotein, a respiratory syncytial virus protein F, a fibronectin A domain, fibrinogen, a measles virus HA protein, and Pam2Cys lipoprotein / lipopeptide (MALP-2). In some embodiments, the immune-cell targeting polypeptide is capable of targeting the ENP to one or more immune cells each selected from the group comprising: a T-cell, a B-cell, a macrophage, a neutrophil, a dendritic cell, optionally, a follicular dendritic cell, an innate lymphoid cell, a mast cell, an eosinophil, a basophil, a megakaryocyte, or a natural killer cell.

[0033] In some embodiments, the cell surface domain and the chimeric protein transmembrane domain are separated by a linker, wherein the linker: is a flexible linker, a rigid linker, or a hybrid linker; is hydrophilic or hydrophobic; is between 1 and 250 amino acids; comprises one or more flexible amino acid residues, optionally about 1 to about 250 flexible amino acid residues, further optionally the flexible amino acid residues comprise glycine, serine, or a combination thereof; and / or comprises 3 repeating amino acid subunits or more.

[0034] In some embodiments, the cell surface domain comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 128 or SEQ ID NO: 130. In some embodiments, the cytoplasmic domain and / or transmembrane domain of the recombinant protein comprise an amino acid sequence having at least 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of the cytoplasmic domain and / or transmembrane domain of the chimeric protein. In some embodiments, (i) the transmembrane domain and the cytoplasmic domain of the recombinant protein and (ii) the transmembrane domain and the cytoplasmic of the chimeric protein each comprise an amino acid sequence having at least 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 129. In some embodiments, the chimeric protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 124, SEQ ID NO: 125, or SEQ ID NO: 126. In some embodiments, the AP of the recombinant protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 40 or SEQ ID NO: 47.

[0035] In some embodiments, one or more of (i) the polynucleotide encoding therecombinant protein or the n polynucleotides each encoding an nth recombinant protein, and (ii) the polynucleotide encoding the chimeric protein or the z polynucleotides each encoding a zth chimeric protein, are present in a same or different nucleic acid molecule. In some embodiments, the amount of (i) the polynucleotide encoding the recombinant protein or the n polynucleotides each encoding an nth recombinant protein; and (ii) the polynucleotide encoding the chimeric protein or the z polynucleotides each encoding a zth chimeric protein, are present in the composition at a molar ratio of about 10: 1, 4: 1, or 1 : 1. In some embodiments, the polynucleotide encoding the recombinant protein and the polynucleotide encoding the chimeric protein are present in the same nucleic acid molecule. In some embodiments, the cell expressing the recombinant protein or the plurality of recombinant proteins and the chimeric protein or plurality of chimeric proteins exhibits at least a 2-fold increase in AP expression at the surface of the cell, relative to a cell expressing a fusion protein or a plurality of fusion proteins comprising an AP and an ERD and does not express the chimeric protein.

[0036] In some embodiments, the nucleic acid composition further comprises a polynucleotide comprising or encoding a tetherin inhibitor. In some embodiments, the tetherin inhibitor is capable of modulating expression, concentration, localization, stability, and / or activity of tetherin. In some embodiments, the tetherin inhibitor comprises a dsRNA, an siRNA, an shRNA, a pre-miRNA, a pri-miRNA, a miRNA, an stRNA, an IncRNA, a piRNA, a snoRNA, or a protein. In some embodiments, one or more of (i) the polynucleotide comprising or encoding the tetherin inhibitor, (ii) the polynucleotide encoding the recombinant protein or the n polynucleotides each encoding an nth recombinant protein, and (iii) the polynucleotide encoding the chimeric protein or the z polynucleotides each encoding a zth chimeric protein, are present in a same or different nucleic acid molecule. In some embodiments, the amount of (i) the polynucleotide comprising or encoding the tetherin inhibitor; and (ii) the polynucleotide encoding the recombinant protein or the n polynucleotides each encoding an nth recombinant protein and / or the polynucleotide encoding the chimeric protein or the z polynucleotides each encoding a zth chimeric protein, are present in the composition at a molar ratio of about 1 : 1, 1 :5 or 1 :25. In some embodiments, the polynucleotide comprising or encoding the tetherin inhibitor and the polynucleotide encoding the recombinant protein are present in the same nucleic acid. In some embodiments, the polynucleotide comprising or encoding the tetherin inhibitor and the polynucleotide encoding the chimeric protein are present in the same nucleic acid. In some embodiments, the tetherin inhibitor comprises or is derived from a viral protein, optionally the virus is HIV-1, HIV-2, SIV, Ebola virus, KSHV, SARS CoV, or SARS-CoV-2. In some embodiments, the tetherin inhibitor comprises HIV-1 Vpu protein, KSHV K5 protein, SARS- CoV-2 ORF7a, HIV-2 Env, Ebola GP, SIV Env, SIV Vpu, SIV Nef, or any portions, variants orderivatives thereof. In some embodiments, the tetherin inhibitor comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 132-134. In some embodiments, the tetherin inhibitor comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 132. In some embodiments, presence or expression of the tetherin inhibitor in the cell results in an increase in ENP production by the cell by at least 2-fold, relative to a cell that does not comprise or express the tetherin inhibitor.

[0037] In some embodiments, the ENPs comprise a lipid bilayer. In some embodiments, the ENPs comprise a lipid bilayer derived from the cell from which the ENP was secreted. In some embodiments, the ERD is located at the C-terminus of the dimerization fusion protein, the adapter fusion protein, the recombinant protein and / or the chimeric protein. In some embodiments, the ERD is capable of interacting with the ESCRT proteins TSG101, NEDD4, and / or ALIX. In some embodiments, the ERD comprises or is derived from a nonhuman protein. In some embodiments, the ERD comprises or is derived from a human protein. In some embodiments, the ERD comprises or is derived from a nonmammalian protein. In some embodiments, the ERD comprises or is derived from a chicken protein, a mouse protein, a lizard protein, a reptile protein, a hamster protein, or a goldfish protein. In some embodiments, the ERD comprises or is derived from the ESCRT and ALIX binding region (EABR) of the human CEP55 protein. In some embodiments, the ERD comprises or is derived from residues 170-213 of the human CEP55 protein. In some embodiments, the ERD comprises or is derived from Syntenin-1, rat Galectin-3 (rGalectin-3), Hrs, and / or CD2AP. In some embodiments, the ERD comprises or is derived from a viral protein. In some embodiments, the ERD comprises or is derived from a fragment of a viral protein. In some embodiments, the ERD comprises or is derived from a retroviral protein, herpes simplex viral protein, vaccinia viral protein, hepadnaviral protein, togaviral protein, flaviviral protein, arenaviral protein, coronaviral protein, orthomyxoviral protein, paramyxoviral protein, bunyaviral protein, bornaviral protein, rhabdoviral protein or filoviral protein. In some embodiments, the ERD comprises or is derived from a Gag protein. In some embodiments, the ERD comprises or is derived from EIAV, HTLV-1, MLV, or MPMV. In some embodiments, the ERD comprises or is derived from EIAV p9, SIV p6 and / or HIV-1 p6; and / or an Ebola protein. In some embodiments, the ERD comprises or is derived from EBOV VP40. In some embodiments, the ERD comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the amino acid sequence of SEQ ID NO: 2.

[0038] In some embodiments, the ERD comprises one or more TSG101 -bindingmotifs, one or more ALIX-binding motifs, one or more Nedd4-recruiting motifs, or any combination thereof. In some embodiments, any two of the one or more TSG101 -binding motifs, the one or more ALIX-binding motifs, or the one or more Nedd4-recruiting motifs are the same or different. In some embodiments, the ERD comprises or is derived from HIV-1 p6 protein or SIV p6 protein. In some embodiments, the ERD comprises or is derived from the p6 protein of HIV-1 isolate ETH2220. In some embodiments, the ERD comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 5-37.

[0039] In some embodiments, the ERD comprises or is derived from a non-human galectin protein. In some embodiments, the ERD comprises or is derived from a rat galectin protein. In some embodiments, the ERD comprising or derived from a non-human galectin protein further comprises a viral-derived ALIX-recruiting motif. In some embodiments, the ERD comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 38-39.

[0040] In some embodiments, the dimerization fusion protein, the adapter fusion protein, the recombinant protein, and / or the chimeric protein comprises an endocytosis-preventing motif (EPM) capable of preventing endocytosis of the dimerization fusion protein, the adapter fusion protein, the recombinant protein, and / or the chimeric protein. In some embodiments, the EPM: tethers the dimerization fusion protein, the adapter fusion protein, the recombinant protein, and / or the chimeric protein to the cytoskeleton, thereby preventing localization to coated pits and endocytosis; enhances ENP assembly, ENP production, and / or ENP secretion; and / or prevents endocytosis of the dimerization fusion protein, the adapter fusion protein, the recombinant protein, and / or the chimeric protein, thereby extending the time the dimerization fusion protein, the adapter fusion protein, the recombinant protein, and / or the chimeric protein remains at the plasma membrane to interact with ESCRT proteins. In some embodiments, the EPM: increases the abundance and / or density of dimerization fusion proteins, adapter fusion proteins, recombinant proteins, and / or chimeric proteins on and / or in the ENP by at least about 2-fold as compared to an ENP comprising a dimerization fusion protein, an adapter fusion protein, a recombinant protein, and / or a chimeric protein that does not comprise the EPM; and / or increases the number of ENPs secreted by a cell by at least about 2-fold as compared to a cell expressing a dimerization fusion protein, an adapter fusion protein, a recombinant protein, and / or a chimeric protein that does not comprise the EPM. In some embodiments, the EPM comprises or is derived from a portion of murine low-affinity gamma Fc region receptor II isoform FcRII-Bl. In some embodiments, the EPM comprises all or a portion of the cytoplasmic tail of FcRII-Bl. In some embodiments, theEPM comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the amino acid sequence of SEQ ID NO: 1. In some embodiments, the dimerization fusion protein, the adapter fusion protein, the recombinant protein, and / or the chimeric protein does not comprise an endocytosis-preventing motif (EPM).

[0041] In some embodiments, the AP, or a portion thereof, is displayed in and / or on the surface of the ENP. In some embodiments, the AP is about 1 amino acid to about 10000 amino acids in length. In some embodiments, the AP comprises or is derived from an antigenic protein associated with a disease or disorder. In some embodiments, the AP comprises or is derived from an immunogenic variant and / or an immunogenic fragment of said antigenic protein. In some embodiments, the AP comprises or is derived from a conserved portion of said antigenic protein. In some embodiments, the AP is present on and / or in the ENP in its natural membrane-associated conformation. In some embodiments, the AP comprises or is derived from at least about 0.01 percent of the full length of said antigenic protein. In some embodiments, the AP comprises or is derived from the full-length surface protein of an infectious agent. In some embodiments, the disease or disorder is an infectious disease or disorder caused by an infectious agent, wherein the AP comprises or is derived from an antigenic protein of said infectious agent, and wherein the antigenic protein of said infectious agent is a pathogenic antigen. In some embodiments, the disease or disorder is a disease associated with expression of a tumor-associated antigen, and the antigenic protein is a tumor-associated antigen. In some embodiments, the disease or disorder is an autoimmune disease or disorder, and the antigenic protein is an autoimmune antigen. In some embodiments, the disease or disorder is an allergic disease or disorder. In some embodiments, the antigenic protein is an allergenic antigen. In some embodiments, the infectious agent is: a bacterium, a fungus, a virus, or a protist; and / or a coronavirus (CoV). In some embodiments, the CoV comprises an alphacoronavirus, a betacoronavirus, a gammacoronavirus, or a deltacoronavirus.

[0042] In some embodiments, the infectious agent is selected from the group comprising Acinetobacter baumannii, Anaplasma genus, Anaplasma phagocytophilum, Ancylostoma braziliense, Ancylostoma duodenale, Arcanobacterium haemolyticum, Ascaris lumbricoides, Aspergillus genus, Astroviridae, Babesia genus, Bacillus anthracis, Bacillus cereus, Bartonella henselae, BK virus, Blastocystis hominis, Blastomyces dermatitidis, Bordetella pertussis, Borrelia burgdorferi, Borrelia genus, Borrelia spp, Brucella genus, Brugia malayi, Bunyaviridae family, Burkholderia cepacia and other Burkholderia species, Burkholderia mallei, Burkholderia pseudomallei, Caliciviridae family, Campylobacter genus, Candida albicans, Candida spp, Chlamydia trachomatis, Chlamydophila pneumoniae, Chlamydophila psittaci, CJD prion, Clonorchis sinensis, Clostridium botulinum, Clostridium difficile, Clostridium perfr ingens,Clostridium perfringens, Clostridium spp, Clostridium tetani, Coccidioides spp, coronaviruses, Corynebacterium diphtherias, Coxiella burnetii, Crimean-Congo hemorrhagic fever virus, Cryptococcus neoformans, Cryptosporidium genus, Cytomegalovirus (CMV), Dengue viruses (DEN-1, DEN-2, DEN-3 and DEN-4), Dientamoeba fragilis, Ebolavirus (EBOV), Echinococcus genus, Ehrlichia chaffeensis, Ehrlichia ewingii, Ehrlichia genus, Entamoeba histolytica, Enterococcus genus, Enterovirus genus, Enteroviruses, mainly Coxsackie A virus and Enterovirus 71 (EV71), Epidermophyton spp, Epstein-Barr Virus (EBV), Escherichia coli O157:H7, 0111 and 0104 :H4, Fasciola hepatica and Fasciola gigantica, FFI prion, Filarioidea superfamily, Filoviruses, Flaviviruses, Francisella tularensis, Fusobacterium genus, Geotrichum candidum, Giardia intestinalis, Gnathostoma spp, GSS prion, Guanarito virus, Haemophilus ducreyi, Haemophilus influenzae, Helicobacter pylori, Henipavirus (Hendra virus Nipah virus), Hepatitis A Virus, Hepatitis B Virus (HBV), Hepatitis C Virus (HCV), Hepatitis D Virus, Hepatitis E Virus, Herpes simplex virus 1 and 2 (HSV-1 and HSV- 2), Histoplasma capsulatum, HIV (Human immunodeficiency virus), Hortaea werneckii, Human bocavirus (HboV), Human herpesvirus 6 (HHV-6) and Human herpesvirus 7 (HHV-7), Human metapneumovirus (hMPV), Human papillomavirus (HPV), Human parainfluenza viruses (HPIV), Japanese encephalitis virus, JC virus, Junin virus, Kingella kingae, Klebsiella granulomatis, Kuru prion, Lassa virus, Legionella pneumophila, Leishmania genus, Leptospira genus, Listeria monocytogenes, Lymphocytic choriomeningitis virus (LCMV), Machupo virus, Malassezia spp, Marburg virus, Measles virus, Metagonimus yokagawai, Microsporidia phylum, Molluscum contagiosum virus (MCV), Mumps virus, Mycobacterium leprae and Mycobacterium lepromatosis, Mycobacterium tuberculosis, Mycobacterium ulcerans, Mycoplasma pneumoniae, Naegleria fowleri, Necator americanus, Neisseria gonorrhoeas, Neisseria meningitidis, Nocardia asteroides, Nocardia spp, Onchocerca volvulus, Orientia tsutsugamushi, Orthomyxoviridae family (Influenza), Paracoccidioides brasiliensis, Paragonimus spp, Paragonimus westermani, ParvovirusBl 9, Pasteurella genus, Plasmodium genus, Pneumocystis jirovecii, Poliovirus, Rabies virus, Respiratory syncytial virus (RSV), Rhinovirus, rhinoviruses, Rickettsia akari, Rickettsia genus, Rickettsia prowazekii, Rickettsia rickettsii, Rickettsia typhi, Rift Valley fever virus, Rotavirus, Rubella virus, Sabia virus, Salmonella genus, Sarcoptes scabiei, SARS coronavirus, Schistosoma genus, Shigella genus, Sin Nombre virus, Hantavirus, Sporothrix schenckii, Staphylococcus genus, Staphylococcus genus, Streptococcus agalactiae, Streptococcus pneumoniae, Streptococcus pyogenes, Strongyloides stercoralis, Taenia genus, Taenia solium, Tick-borne encephalitis virus (TBEV), Toxocara canis or Toxocara cati, Toxoplasma gondii, Treponema pallidum, Trichinella spiralis, Trichomonas vaginalis, Trichophyton spp, Trichuristrichiura, Trypanosoma brucei, Trypanosoma cruzi, Ureaplasma urealyticum, Varicella zoster virus (VZV), Variola major or Variola minor, vCJD prion, Venezuelan equine encephalitis virus, Vibrio cholerae. West Nile virus, Western equine encephalitis virus, Wuchereria bancrofli. Yellow fever virus, Yersinia enter ocolitica, Yersinia pestis, and Yersinia pseudotuberculosis.

[0043] In some embodiments, the AP comprises a membrane protein (e.g., a multispan transmembrane protein). In some embodiments, the AP is not configured to be a soluble protein. In some embodiments, the AP does not comprise one or more mutations configured to enhance its solubility and / or stability. In some embodiments, the AP does not comprise a transmembrane domain and / or is a soluble protein, and the dimerization fusion protein and the adapter fusion protein comprises a transmembrane domain (TD). In some embodiments, the transmembrane domain comprises or is derived from a human transmembrane protein or nonhuman transmembrane protein. In some embodiments, the TD comprises or is derived from a natural protein, a recombinant protein, and / or synthetic protein (e.g., a synthetic protein comprising predominantly hydrophobic residues).

[0044] In some embodiments, the population of ENPs comprise one or more homotypic ENPs. In some embodiments, the plurality of dimerization fusion proteins or the plurality of recombinant proteins of a homotypic ENP are the same as each other with respect to the AP. In some embodiments, a homotypic ENP thereby does not display a plurality of disparate AP. In some embodiments, the population of ENPs comprise one or more heterotypic ENPs, wherein at least two of the dimerization fusion proteins or recombinant proteins of a heterotypic ENP are different from each other with respect to the AP, and a heterotypic ENP thereby displays a plurality of disparate AP. In some embodiments, the population of ENPs comprise a mixture of two or more homotypic ENPs that differ from each other with respect to the AP of the plurality of dimerization fusion proteins or the plurality of recombinant proteins present in said two or more homotypic ENPs, and the population of ENPs thereby displays a plurality of disparate AP. In some embodiments, the population of ENPs comprise a mixture of two or more heterotypic ENPs that differ from each other with respect to the AP of the plurality of dimerization fusion proteins or the plurality of recombinant proteins of said two or more heterotypic ENPs. In some embodiments, heterotypic ENPs are capable of eliciting heterologous antibody responses against an additional infectious agent, and said heterotypic ENPs do not display AP derived from said additional infectious agent. In some embodiments, the plurality of disparate AP comprises: between about 2 and about 500 antigenic polypeptides that differ from each other; AP of a same protein type; and / or AP of different protein types. In some embodiments, the same ENP comprises the AP derived from two or more strains of the same family, same genus, and / or same species, of infectious agent. In some embodiments, the plurality of disparate AP has a sequence identity ofabout, at least, or at least about 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% with one another.

[0045] In some embodiments, the plurality of disparate AP comprise a plurality of coronavirus (CoV) antigens. In some embodiments, the plurality of CoV antigens comprises a first CoV antigen of a first CoV and a second CoV antigen of a second CoV that is different from the first CoV. In some embodiments, the plurality of CoV antigens comprise a CoV spike protein (S protein) or a portion thereof, a CoV envelope protein (E protein) or a portion thereof, a CoV nucleocapsid protein (N protein) or a portion thereof, a CoV hemagglutinin-esterase protein (HE protein) or a portion thereof, a CoV papain-like protease or a portion thereof, a CoV 3 CL protease or a portion thereof, a CoV membrane protein (M protein) or a portion thereof, or a combination thereof. In some embodiments, the plurality of CoV antigens comprise a CoV S protein or a portion thereof. In some embodiments, the first CoV antigen, the second CoV antigen, or both comprise a CoV S protein or a portion thereof. In some embodiments, the number of the first CoV antigen molecules and the number of the second CoV antigen molecules are in a ratio from 1 : 100 to 100: 1. In some embodiments, the plurality of CoV antigens comprise three, four, five, size seven, or eight CoV antigens, each of a CoV different from one another. In some embodiments, the plurality of CoV antigens further comprise at least a third CoV antigen of a third CoV and a fourth CoV antigen of a fourth CoV. In some embodiments, the first, second, third and fourth Co Vs are different from one another.

[0046] In some embodiments, the plurality of disparate AP comprise at least m pathogenic antigens of an mth infectious agent, wherein m is an integer greater than 2. In some embodiments, each mth pathogenic antigen is different from one another. In some embodiments, m is an integer greater than 50. In some embodiments, the plurality of disparate AP comprise one or more of a 1st pathogenic antigen (PA) of a 1st infectious agent (IA), a 2nd PA of a 2nd IA, a 3rd PA of a 3rd I A, a 4th PA of a 4th I A, a 5 th PA of a 5 th IA, a 6th PA of a 6th IA, a 7th PA of a 7th IA, a 8th PA of a 8th IA, a 9th PA of a 9th IA, a 10th PA of a 10th IA, a 11th PA of a 11th IA, a 12th PA of a 12th IA, a 13th PA of a 13th IA, a Mth PA of a Mth IA, a 15th PA of a 15thIA, a 16th PA of a 16th IA, a 17th PA of a 17th IA, a 18th PA of a 18th IA, a 19th PA of a 19thIA, a 20th PA of a 20th IA, a 21st PA of a 21st IA, a 22nd PA of a 22nd IA, a 23rd PA of a 23rd IA, a 24th PA of a 24th IA, a 25th PA of a 25th IA, a 26th PA of a 26th IA, a 27th PA of a 27thIA, a 28th PA of a 28th IA, a 29th PA of a 29th IA, a 30th PA of a 30th IA, a 31st PA of a 31stIA, a 32nd PA of a 32nd IA, a 33rd PA of a 33rd IA, a 34th PA of a 34th IA, a 35th PA of a 35thIA, a 36th PA of a 36th IA, a 37th PA of a 37th IA, a 38th PA of a 38th IA, a 39th PA of a 39thI A, a 40th PA of a 40th I A, a 41st PA of a 41st I A, a 42nd PA of a 42nd I A, a 43 rd PA of a 43 rdIA, a 44th PA of a 44th IA, a 45th PA of a 45th IA, a 46th PA of a 46th IA, a 47th PA of a 47thIA, a 48th PA of a 48th IA, a 49th PA of a 49th IA, and a 50th PA of a 50th IA, wherein the 1st, 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, 9th, 10th, 11th, 12th, 13th, 14th, 15th, 16th, 17th, 18th, 19th, 20th, 21st, 22nd, 23rd, 24th, 25th, 26th, 27th, 28th, 29th, 30th, 31st, 32nd, 33rd, 34th, 35th, 36th, 37th, 38th, 39th, 40th, 41st, 42nd, 43rd, 44th, 45th, 46th, 47th, 48th, 49th, and 50th pathogenic antigens are different from one another. In some embodiments, the 1st, 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, 9th, 10th, 11th, 12th, 13th, 14th, 15th, 16th, 17th, 18th, 19th, 20th, 21st, 22nd, 23rd, 24th, 25th, 26th, 27th, 28th, 29th, 30th, 31st, 32nd, 33rd, 34th, 35th, 36th, 37th, 38th, 39th, 40th, 41st, 42nd, 43rd, 44th, 45th, 46th, 47th, 48th, 49th, and 50th infectious agents are different from one another. In some embodiments, the plurality of disparate AP comprise the plurality of CoV antigens, a plurality of influenza antigens, and / or a plurality of HIV antigens.

[0047] In some embodiments, one or more of the plurality of CoV antigens are of CoVs in the genus of Alpha-CoV and / or Beta-CoV. In some embodiments, each of the plurality of CoV antigens are of CoVs in the genus of Beta-CoV. In some embodiments, the plurality of CoV antigens are of CoVs in the subgenus of Sarbecovirus; the first CoV and the second CoV are in the genus of Beta-CoV (e.g., in the subgenus of Sarbecovirus. In some embodiments, plurality of CoV antigens are of CoVs selected from the group consisting of: SARS-CoV, SARS-CoV-2, WIV1, SHC014, Rfl, RmYN02, pangl7, RaTG13, Rs4081, LYRal l, HKU3, Yunnan2011, BtKY72, BM48-31, WIV16, Khosta-1, and Khosta-2. In some embodiments, first CoV, the second CoV, or both are selected from the group consisting of: SARS-CoV, SARS-CoV-2, WIV1, SHC014, Rfl, RmYN02, pangl7, RaTG13, Rs4081, LYRal l, HKU3, Yunnan2011, BtKY72, BM48-31, WIV16, Khosta-1, and Khosta-2. In some embodiments, CoV is selected from a species or subspecies of SARS-CoV, SARS-CoV-1, SARS-CoV-2, MERS-CoV, SL-CoV-WIVl, HKU4, HKU5, HCoV-OC43, HCoV-HKUl, HKU9, HKU3, HKU8, HKU24, NL63, SHC014, 229E and / or SARS-CoV-2 variants B.1.351, B.l.1.7, P. l, B.1.617.2, B.1.1.529, BA.l, BA.1.1, BA.2, BA.3, BA.4, BA.5 and other descendent lineages. In some embodiments, CoV is selected from a species or subspecies of Embecovirus, Sarbecovirus, Merbecovirus, Nobevovirus, Hibecovirus, SARSr-CoV, MERS-CoV, or any combination thereof. In some embodiments, CoV is selected from a beta-CoV from the sarbe-, embeco-, merbeco-, and nobecovirus lineages. In some embodiments, the CoV is selected from a a sarbecovirus strain (e.g., SARS, LYRal l, Rfl, Rs4081, BtKY72, and / or BM48-31); a merbecovirus strain (e.g., HKU4, HKU5, HKU25, BtCoV- Vs-CoVl, MERS-related NL13845, and MERS-related NL 140422); and / or an embecovirus strain (e.g., HKU1, Rat CoV Parker, PHEV, Equine CoV, Rodent CoV, and Longquan Rat CoV).

[0048] In some embodiments, the ENPs comprise at least about 2-fold more of the AP and / or are at least as immunogenic as compared to a multi-component nanoparticle approach, (e.g., as compared to a SpyCatcher-based nanoparticle approach or a lentiviral Gag-basedapproach). In some embodiments, the multi-component nanoparticle approach comprises two or more separate polypeptides. In some embodiments, the ENPs comprise at least about 2-fold more of the AP, an at least about 2-fold higher density of the AP, and / or are at least as immunogenic, as compared to a nanoparticle approach that does not comprise the ERD, (e.g., as compared to a SpyCatcher-based or Gag-based nanoparticle approach). In some embodiments, the nucleic acid composition does not comprise a polynucleotide encoding SpyTag or lentiviral Gag; and / or the ENPs do not comprise SpyTag or lentiviral Gag.

[0049] In some embodiments, the ENPs have one or more dimensions of a eukaryotic virus; less than about 10% of the ENPs of the population of ENPs have a particle size smaller than about 10 nm; less than about 10% of the ENPs of the population of ENPs have a particle size exceeding about 80 nm; the average diameter of the ENPs of the population of ENPs range from about 5 nm to about 80 nm, from about 15 nm to about 50 nm, or from about 20 nm to about 40 nm; and / or the average diameter of the ENPs of the population of ENPs is about 10 nm, about 12 nm, about 14 nm, about 16 nm, about 18 nm, about 20 nm, about 22 nm, about 24 nm, about 26 nm, about 28 nm, about 30 nm, about 32 nm, about 34 nm, about 36 nm, about 38 nm, about 40 nm, about 42 nm, about 44 nm, about 46 nm, about 48 nm, or about 50 nm. In some embodiments, the average is the mean, median or mode. In some embodiments, the mean is the arithmetic mean, geometric mean, and / or harmonic mean. In some embodiments, the ENPs: have a minimum diameter of about 10 nm, about 12 nm, about 14 nm, about 16 nm, about 18 nm, about 20 nm, about 22 nm, about 24 nm, about 26 nm, about 28 nm, about 30 nm, about 32 nm, about 34 nm, about 36 nm, about 38 nm, about 40 nm, about 42 nm, about 44 nm, about 46 nm, about 48 nm, or about 50 nm; have a maximum diameter of about 10 nm, about 12 nm, about 14 nm, about 16 nm, about 18 nm, about 20 nm, about 22 nm, about 24 nm, about 26 nm, about 28 nm, about 30 nm, about 32 nm, about 34 nm, about 36 nm, about 38 nm, about 40 nm, about 42 nm, about 44 nm, about 46 nm, about 48 nm, about 50 nm, about 52 nm, about 54 nm, about 56 nm, about 58 nm, about 60 nm, about 62 nm, about 64 nm, about 66 nm, about 68 nm, about 70 nm, about 72 nm, about 74 nm, about 76 nm, about 78 nm, or about 80 nm. In some embodiments, the average diameter of the ENPs of the population of ENPs can exceed 80 nm. In some embodiments, the ENPs are derived from cell cultures transiently transfected with the nucleic acid composition, optionally derived via ultracentrifugation and / or size exclusion chromatography, further optionally ultracentrifugation on a 20% sucrose cushion, optionally transfected via calcium phosphate transfection, DEAE-dextran mediated transfection, cationic lipid-mediated transfection, electroporation, electrical nuclear transport, chemical transduction, electrotransduction, Lipofectamine-mediated transfection, Effectene-mediated transfection, lipid nanoparticle (LNP)-mediated transfection, or any combination thereof.

[0050] In some embodiments, storage of the ENPs at 4°C for at least three months reduces immunogenicity less than about 50 percent; the composition is stable for at least about 2 weeks, about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, or about 1 year, after storage as a liquid at a temperature of about 4°C; and / or at least about 70%, 75%, 80%, 85%, 90% or 95% of the ENPs are immunogenic at least 1 month after storage as a liquid at a temperature of about 5°C. In some embodiments, the nucleic acid composition is complexed or associated with one or more lipids or lipid-based carriers, thereby forming liposomes, lipid nanoparticles (LNPs), lipoplexes, and / or nanoliposomes, e.g., encapsulating the nucleic acid composition.

[0051] In some embodiments, the nucleic acid composition is, comprises, or further comprises, one or more vectors. In some embodiments, at least one of the one or more vectors is a viral vector, a plasmid, a transposable element, a naked DNA vector, a lipid nanoparticle (LNP), or any combination thereof. In some embodiments, the viral vector is an AAV vector, a lentivirus vector, a retrovirus vector, an adenovirus vector, a herpesvirus vector, a herpes simplex virus vector, a cytomegalovirus vector, a vaccinia virus vector, a MVA vector, a baculovirus vector, a vesicular stomatitis virus vector, a human papillomavirus vector, an avipox virus vector, a Sindbis virus vector, a VEE vector, a Measles virus vector, an influenza virus vector, a hepatitis B virus vector, an integration-deficient lentivirus (IDLV) vector, or any combination thereof. In some embodiments, the transposable element is piggybac transposon or sleeping beauty transposon.

[0052] In some embodiments, the polynucleotide(s) encoding the dimerization fusion protein, the adapter fusion protein, the recombinant protein, and / or the chimeric protein are comprised in the one or more vectors. In some embodiments, the polynucleotide(s) encoding any of the dimerization fusion protein, the adapter fusion protein, the recombinant protein, and / or the chimeric protein are comprised in the same vector and / or different vectors. In some embodiments, the polynucleotide(s) encoding any of the dimerization fusion protein, the adapter fusion protein, the recombinant protein, and / or the chimeric protein are situated on the same nucleic acid and / or different nucleic acids. In some embodiments, the one or more vectors is a DNA vaccine. In some embodiments, the polynucleotide(s) encoding any of the dimerization fusion protein, the adapter fusion protein, the recombinant protein, and / or the chimeric protein are operably linked to one or more promoters capable of inducing transcription of said polynucleotide(s). In some embodiments, the DNA vaccine is a plasmid-based DNA vaccine, a minicircle-based DNA vaccine, a bacmid-based DNA vaccine, a minigene-based DNA vaccine, a ministring DNA (linear covalently closed DNA vector) vaccine, a closed-ended linear duplex DNA (CELiD or ceDNA) vaccine, a doggybone™ DNA vaccine, a dumbbell shaped DNA vaccine, or a minimalistic immunological-defmed gene expression (MIDGE)-vector DNA vaccine. In some embodiments,the DNA vaccine elicits at least 2-fold higher neutralizing antibody responses against an infectious agent as compared to a DNA vaccine that encodes the AP only.

[0053] In some embodiments, the promoter comprises a ubiquitous promoter, an inducible promoter, a tissue-specific promoter and / or a lineage-specific promoter. In some embodiments, the ubiquitous promoter is selected from the group comprising a cytomegalovirus (CMV) immediate early promoter, a CMV promoter, a viral simian virus 40 (SV40) (e.g., early or late), a Moloney murine leukemia virus (MoMLV) LTR promoter, a Rous sarcoma virus (RS V) LTR, an RSV promoter, a herpes simplex virus (HSV) (thymidine kinase) promoter, H5, P7.5, and Pl l promoters from vaccinia virus, an elongation factor 1 -alpha (EFla) promoter, early growth response 1 (EGR1), ferritin H (FerH), ferritin L (FerL), Glyceraldehyde 3 -phosphate dehydrogenase (GAPDH), eukaryotic translation initiation factor 4A1 (EIF4A1), heat shock 70 kDa protein 5 (HSPA5), heat shock protein 90 kDa beta, member 1 (HSP90B 1), heat shock protein 70 kDa (HSP70), P-kinesin (P-KIN), the human ROSA 26 locus, a Ubiquitin C promoter (UBC), a phosphoglycerate kinase- 1 (PGK) promoter, 3 -phosphoglycerate kinase promoter, a cytomegalovirus enhancer, human P-actin (HBA) promoter, chicken P-actin (CBA) promoter, a CAG promoter, a CASI promoter, a CBH promoter, or any combination thereof; the polynucleotide(s) encoding any of the dimerization fusion protein, the adapter fusion protein, the recombinant protein, and / or the chimeric protein are operably linked to a tandem gene expression element. In some embodiments, the tandem gene expression element is an internal ribosomal entry site (IRES), foot-and-mouth disease virus 2A peptide (F2A), equine rhinitis A virus 2A peptide (E2A), porcine teschovirus 2A peptide (P2A) or Thosea asigna virus 2A peptide (T2A), or any combination thereof; and / or the polynucleotide(s) encoding any of the dimerization fusion protein, the adapter fusion protein, the recombinant protein, and / or the chimeric protein comprises a transcript stabilization element. In some embodiments, the transcript stabilization element comprises woodchuck hepatitis post-translational regulatory element (WPRE), bovine growth hormone polyadenylation (bGH-polyA) signal sequence, human growth hormone polyadenylation (hGH-polyA) signal sequence, or any combination thereof.

[0054] In some embodiments, the nucleic acid composition is or comprises mRNA. In some embodiments, the mRNA is formulated in a lipid nanoparticle (LNP). In some embodiments, the mRNA comprises: a 5' untranslated region (UTR), a 3' UTR, and / or a cap; one or more modified nucleotides selected from the group comprising pseudouridine, N-l-methyl- pseudouridine, 2-aminoadenosine, 2-thiothymidine, inosine, pyrrolo-pyrimidine, 3 -methyl adenosine, 5-methylcytidine, C-5 propynyl-cytidine, C-5 propynyl-uridine, 2-aminoadenosine, C5-bromouridine, C5-fluorouridine, C5-iodouridine, C5-propynyl-uridine, C5-propynyl-cytidine, C5-methylcytidine, 2-aminoadenosine, 7-deazaadenosine, 7-deazaguanosine, 8-oxoadenosine, 8-oxoguanosine, 0(6)-methylguanine, and 2-thiocytidine; and / or a modified nucleotide in place of one or more uridines. In some embodiments, the modified nucleoside is selected from pseudouridine (y), N 1-methyl-pseudouridine (m IT), and 5-methyl-uridine (m5U).

[0055] In some embodiments, the LNP comprises: one or more of an ionizable cationic lipid, a non-cationic lipid, a sterol, and a PEG-modified lipid. In some embodiments, the noncationic lipid is a neutral lipid; 0.5-15 mol% PEG-modified lipid, 5-25 mol% non-cationic lipid, 25-55 mol% sterol, and 20-60 mol% ionizable cationic lipid; and / or 40-55 mol% ionizable cationic lipid, 5-15 mol% neutral lipid, 35-45 mol% sterol, and 1-5 mol% PEG-modified lipid. In some embodiments, the LNP comprises: 47 mol% ionizable cationic lipid, 11.5 mol% neutral lipid, 38.5 mol% sterol, and 3.0 mol% PEG-modified lipid; 48 mol% ionizable cationic lipid, 11 mol% neutral lipid, 38.5 mol% sterol, and 2.5 mol% PEG-modified lipid; 49 mol% ionizable cationic lipid, 10.5 mol% neutral lipid, 38.5 mol% sterol, and 2.0 mol% PEG-modified lipid; 50 mol% ionizable cationic lipid, 10 mol% neutral lipid, 38.5 mol% sterol, and 1.5 mol% PEG- modified lipid; or 51 mol% ionizable cationic lipid, 9.5 mol% neutral lipid, 38.5 mol% sterol, and 1.0 mol% PEG-modified lipid. In some embodiments, the ionizable cationic lipid is heptadecan- 9-yl 8 ((2 hydroxy ethyl)(6 oxo 6-(undecyloxy)hexyl)amino)octanoate; the neutral lipid is 1,2 distearoyl-sn-glycero-3 phosphocholine (DSPC); the sterol is cholesterol; and / or the PEG- modified lipid is l-monomethoxypolyethyleneglycol-2,3-dimyristylglycerol with polyethylene glycol of average molecular weight 2000 (PEG2000 DMG). In some embodiments, the wt / wt ratio of lipid to mRNA is from about 1 : 100 to about 100: 1.

[0056] In some embodiments, (i) the nucleic acid composition comprises one or more polynucleotides encoding immunostimulatory agents; and / or (ii) the population of ENPs comprises one or more immunostimulatory agents. In some embodiments, the immunostimulatory agents are selected from the group comprising toll-like receptor (TLR) agonists, cytokine receptor agonists, CD40 agonists, Fc receptor agonists, CpG-containing nucleic acids, complement receptor agonists, or any combination thereof. In some embodiments, the TLR agonist is a TLR- 1 agonist, TLR-2 agonist, TLR-3 agonist, TLR-4 agonist, TLR-5 agonist, TLR-6 agonist, TLR-7 agonist, TLR-8 agonist, TLR-9 agonist, and / or TLR- 10 agonist; the Fc receptor agonist is a Fc- gamma receptor agonist; the complement receptor agonist binds to CD21 or CD35; the complement receptor agonist induces endogenous complement opsonization of the ENP; the cytokine receptor agonist is a cytokine; and / or the cytokine receptor agonist is a small molecule, antibody, fusion protein, or aptamer.

[0057] In some embodiments, the composition further comprises Tris buffer, sucrose, and / or sodium acetate; and / or further comprises an adjuvant. In some embodiments, the adjuvant is selected from the group comprising aluminum hydroxide, alhydrogel, AddaVax, MF59, AS03,Freund’s adjuvant, Montanide ISA51, CpG, Poly I:C, glucopyranosyl lipid A, flagellin, resiquimod, or any combination thereof. In some embodiments, the composition is a lyophilized composition. In some embodiments, the lyophilized composition has a water content of less than about 10%. In some embodiments, the composition is formulated or is to be formulated: as a liquid, a solid, or a combination thereof; for injection; for intramuscular administration, intranasal administration, transdermal administration, aerosol delivery, nasal delivery, vaginal delivery, rectal delivery, buccal delivery, ocular delivery, local delivery, topical delivery, intracistemal delivery, intraperitoneal delivery, oral delivery, intramuscular injection, intravenous injection, subcutaneous injection, intranodal injection, intratumoral injection, intraperitoneal injection, intradermal injection; and / or as particles. In some embodiments, the particles are iron oxide particles, liposomes, micelles, polymer complexes, cationic peptide nanoemulsions, virus-like particles (VLPs), lipid nanoparticles (LNP) and / or lipoplex (LPX) particles. In some embodiments, the nucleic acid composition and the LNP-forming components are in separate vials. In some embodiments, the composition is a pharmaceutical composition. In some embodiments, the pharmaceutical composition further comprises one or more pharmaceutically acceptable carriers, diluents and / or excipients.

[0058] The composition can comprise instructions for use of the composition for: stimulating an immune response in a subject in need thereof; treating or preventing a disease or disorder caused by an infectious agent in a subject in need thereof; and / or treating or preventing a CoV infection in a subject in need thereof. Disclosed herein include kits. In some embodiments, the kit comprises any of the compositions of the disclosure. Also disclosed herein include a cell or population of cells comprising any of the nucleic acid compositions disclosed herein. Provided here are methods of stimulating an immune response in a subject in need thereof. In some embodiments, the method comprises: administering to the subject a pharmaceutically effective amount of any of the compositions of the disclosure, thereby stimulating an immune response in the subject. Provided here are methods of treating or preventing a disease or disorder in a subject in need thereof. In some embodiments, the method comprises: administering to the subject a pharmaceutically effective amount of any of the compositions disclosed herein, thereby treating or preventing the disease or disorder in the subject, optionally the disease or disorder is a disease or disorder caused by an infectious agent. In some embodiments, the disease or disorder caused by an infectious agent is a disease or disorder caused by a coronavirus (CoV) infection.

[0059] Disclosed herein include methods for treating or preventing a coronavirus (CoV) infection in a subject in need thereof. In some embodiments, the method comprises: administering to the subject a pharmaceutically effective amount of any of the compositions of the disclosure, thereby treating or preventing the CoV infection in the subject.

[0060] In some embodiments, immunogenic levels of any of the dimerization fusion protein, the adapter fusion protein, the recombinant protein, and / or the chimeric protein, and / or ENP are produced in serum of the subject at about 1 hour to about 6 months post administration of the composition. In some embodiments, a neutralizing antibody titer of about 50 to about 100000 half-maximal inhibitory dilutions (ID50s values) is produced in the serum of the subject at about 1 hour to about 6 months post administration of the composition. In some embodiments, the composition elicits at least about 2-fold less off-target immune responses against undesired epitopes as compared to a non-enveloped NP -based composition. In some embodiments, said undesired epitopes comprise the NP scaffold of said non-enveloped NP -based composition. In some embodiments, the method comprises administering to the subject at least two doses of the composition. In some embodiments, a second dose of the composition is administered to the subject at least 14 days after a first dose of the composition is administered to the subject.

[0061] In some embodiments, administering the composition induces neutralizing responses against: the infectious agent(s) from which the antigenic polypeptide(s) are derived; and / or additional infectious agent(s) from which the antigenic polypeptide(s) are not derived, e.g., different from the 1st, 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, 9th, 10th, 11th, 12th, 13th, 14th, 15th, 16th, 17th, 18th, 19th, 20th, 21st, 22nd, 23rd, 24th, 25th, 26th, 27th, 28th, 29th, 30th, 31st, 32nd, 33rd, 34th, 35th, 36th, 37th, 38th, 39th, 40th, 41st, 42nd, 43rd, 44th, 45th, 46th, 47th, 48th, 49th, and / or 50th infectious agent. In some embodiments, administering the composition induces neutralizing responses against: the coronaviruses the plurality of coronavirus antigens are of; coronaviruses different from the 1st, 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, 9th, 10th, 11th, 12th, 13th, 14th, 15th, 16th, 17th, 18th, 19th, 20th, 21st, 22nd, 23rd, 24th, 25th, 26th, 27th, 28th, 29th, 30th, 31st, 32nd, 33rd, 34th, 35th, 36th, 37th, 38th, 39th, 40th, 41st, 42nd, 43rd, 44th, 45th, 46th, 47th, 48th, 49th, and / or 50th CoV; and / or additional coronaviruses different from the coronaviruses the plurality of coronavirus antigens are of.

[0062] In some embodiments, administering the composition results in treating or preventing: infection caused by a coronavirus different from the first coronavirus and the second coronavirus; infection caused by additional coronaviruses different from the coronaviruses the plurality of coronavirus antigens are of; infection caused by the coronaviruses the plurality of coronavirus antigens are of; the disease or disorder caused by a coronavirus different from the first coronavirus and the second coronavirus; the disease or disorder caused by additional coronaviruses different from the coronaviruses the plurality of coronavirus antigens are of; and / or the disease or disorder caused by the coronaviruses the plurality of coronavirus antigens are of.

[0063] In some embodiments, the composition elicits an at least 2-fold higher neutralizing antibody titer as compared to an approach comprising administration of (i) a solubleversion of the AP, and / or (ii) an mRNA vaccine encoding the AP and not encoding the ERD; and / or the composition elicits an at least as high neutralizing antibody titer as compared to an approach comprising administration of a protein-based nanoparticle presenting the AP. In some embodiments, administration of the composition elicits protective and long-lasting immunity against the infectious agent(s) and variants thereof. In some embodiments, an at least as low dose of the composition is needed to generate a comparable immune response as compared to an approach comprising administration of a protein-based nanoparticle presenting the AP; and / or an at least about 2-fold lower dose of the composition is needed to generate a comparable immune response as compared to an approach comprising administration of (i) a soluble version of the AP, and / or (ii) an mRNA vaccine encoding the AP and not encoding the ERD.

[0064] In some embodiments, following administration of the first dose or the second dose of the composition, the composition induces: an at least as potent serum neutralizing titers against the infectious agent or variants thereof as compared to an approach comprising administration of a protein-based nanoparticle presenting the AP. In some embodiments, the composition comprises a population of ENPs; an at least 2-fold more potent serum neutralizing titers against the infectious agent or variants thereof as compared to an approach comprising administration of a soluble version of the AP. In some embodiments, the composition comprises a population of ENPs; and / or an at least 2-fold more potent serum neutralizing titers against the infectious agent or variants thereof as compared to an approach comprising administration of an mRNA vaccine encoding the AP and not encoding the ERD. In some embodiments, the composition comprises an mRNA vaccine encoding a dimerization fusion protein or recombinant protein comprising the AP. In some embodiments, potency of serum neutralizing titers are measured by geometric means for serum half-maximal inhibitory dilutions (ID50s values) against the infectious agent or variants thereof, optionally about 1 day to about 6 months after administration of a first dose of the composition or about 1 day to about 6 months after administration of a second dose of the composition.

[0065] In some embodiments, the subject: is a human subject; is a newborn or infant of an age of not more than 3 years, of not more than 2 years, of not more than 1.5 years, of not more than 1 year (12 months), of not more than 9 months, 6 months or 3 months, or is between 6 months and 2 years; is immunocompromised, has a pulmonary disease, and / or is 65 years of age or older; has a chronic pulmonary disease, optionally chronic obstructive pulmonary disease (COPD) or asthma; and / or has an underlying comorbid condition, optionally selected from heart disease, diabetes, and lung disease.

[0066] In some embodiments, the composition is administered in an effective amount to: induce a robust antibody response against the AP in the subject. In some embodiments, a robustantibody response comprises a neutralizing antibody response. In some embodiments, a robust antibody response comprises Fc domain effector functions that recruit immune cells to infected cells. In some embodiments, said immune cells are macrophages, neutrophils, and / or natural killer cells. In some embodiments, said recruitment induces antibody-dependent cellular cytotoxicity (ADCC) and / or antibody-dependent cellular phagocytosis (ADCP); elicit a robust CD4 and / or CD8 T cell response against the AP in the subject; and / or elicit a balanced Thl / Th2 response against the AP in the subject. In some embodiments, the composition is: (i) co-administered with an adjuvant; or (ii) not co-administered with an adjuvant.

[0067] In some embodiments, the disease or disorder is a blood disease, an immune disease, a neurological disease or disorder, a cancer, an infectious disease, a genetic disease, a disorder caused by aberrant mtDNA, a metabolic disease, a disorder caused by aberrant cell cycle, a disorder caused by aberrant angiogenesis, a disorder cause by aberrant DNA damage repair, or any combination thereof. In some embodiments, the disease or disorder is a solid tumor. In some embodiments, the disease or disorder is an infectious disease selected from the group consisting of an Acute Flaccid Myelitis (AFM), Anaplasmosis, Anthrax, Babesiosis, Botulism, Brucellosis, Campylobacteriosis, Carbapenem-resistant Infection, Chancroid, Chikungunya Virus Infection, Chlamydia, Ciguatera, Difficile Infection, Perfringens, Coccidioidomycosis fungal infection, coronavirus infection, Covid-19 (SARS-CoV-2), Creutzfeldt- Jacob Disease / transmissible spongiform encephalopathy, Cryptosporidiosis (Crypto), Cyclosporiasis, Dengue 1,2,3 or 4, Diphtheria, E. coli infection / Shiga toxin-producing (STEC), Eastern Equine Encephalitis, Hemorrhagic Fever (Ebola), Ehrlichiosis, Encephalitis, Arboviral or parainfectious, Non-Polio Enterovirus, D68 Enterovirus(EV-D68), Giardiasis, Glanders, Gonococcal Infection, Granuloma inguinale, Haemophilus Influenza disease Type B (Hib or H-flu), Hantavirus Pulmonary Syndrome (HPS), Hemolytic Uremic Syndrome (HUS), Hepatitis A (Hep A), Hepatitis B (Hep B), Hepatitis C (Hep C), Hepatitis D (Hep D), Hepatitis E (Hep E), Herpes, Herpes Zoster (Shingles), Histoplasmosis infection, Human Immunodeficiency Virus / AIDS (HIV / AIDS), Human Papillomavirus (HPV), Influenza (Flu), Legionellosis (Legionnaires Disease), Leprosy (Hansens Disease), Leptospirosis, Listeriosis (Listeria), Lyme Disease, Lymphogranuloma venereum infection (LGV), Malaria (e.g., Plasmodium falciparum (Pf), Plasmodium knowlesi (Pk), Plasmodium ovale (Po), Plasmodium simiovale (Ps), and Plasmodium vivax (Pv). In preferred embodiments, the Malaria parasite is Plasmodium falciparum (Pf), Plasmodium malar iae (Pm). Plasmodium ovale curtisi (Poc), Plasmodium ovale wallikeri (Pow), Plasmodium berghei (Pb)), Measles, Melioidosis, Meningitis (Viral), Meningococcal Disease (Meningitis (Bacterial)), Middle East Respiratory Syndrome Coronavirus (MERS-CoV), Mumps, Norovirus, Pediculosis, Pelvic Inflammatory Disease (PID), Pertussis (Whooping Cough), Plague (Bubonic,Septicemic, Pneumonic), Pneumococcal Disease (Pneumonia), Poliomyelitis (Polio), Powassan, Psittacosis, Pthiriasis, Pustular Rash diseases (Small pox, monkeypox, cowpox), Q-Fever, Rabies, Rickettsiosis (Rocky Mountain Spotted Fever), Rubella (German Measles), Salmonellosis gastroenteritis (Salmonella), Scabies, Scombroid, Sepsis, Severe Acute Respiratory Syndrome (SARS), Shigellosis gastroenteritis (Shigella), Smallpox, Staphyloccal Infection Methicillin- resistant (MRSA), Staphylococcal Food Poisoning Enterotoxin B Poisoning (Staph Food Poisoning), Saphylococcal Infection Vancomycin Intermediate (VISA), Staphylococcal Infection Vancomycin Resistant (VRSA), Streptococcal Disease Group A (invasive) (Strep A (invasive), Streptococcal Disease, Group B (Strep-B), Streptococcal Toxic-Shock Syndrome STSS Toxic Shock, Syphilis (primary, secondary, early latent, late latent, congenital), Tetanus Infection, Trichomoniasis, Trichonosis Infection, Tuberculosis (TB), Tuberculosis Latent (LTBI), Tularemia, Typhoid Fever Group D, Vaginosis, Varicella (Chickenpox), Vibrio cholerae (Cholera), Vibriosis (Vibrio), Ebola Virus Hemorrhagic Fever, Lasa Virus Hemorrhagic Fever, Marburg Virus Hemorrhagic Fever, West Nile Virus, Yellow Fever, Yersenia, and Zika Virus Infection. In some embodiments, the disease is associated with expression of a tumor-associated antigen. In some embodiments, the disease or disorder is the disease associated with expression of a tumor antigen-associated is selected from the group consisting of a proliferative disease, a precancerous condition, a cancer, and a non-cancer related indication associated with expression of the tumor antigen; the cancer is selected from the group consisting of colon cancer, rectal cancer, renal-cell carcinoma, liver cancer, non-small cell carcinoma of the lung, cancer of the small intestine, cancer of the esophagus, melanoma, bone cancer, pancreatic cancer, skin cancer, cancer of the head or neck, cutaneous or intraocular malignant melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, stomach cancer, testicular cancer, uterine cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, Hodgkin's Disease, non-Hodgkin lymphoma, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue, cancer of the urethra, cancer of the penis, solid tumors of childhood, cancer of the bladder, cancer of the kidney or ureter, carcinoma of the renal pelvis, neoplasm of the central nervous system (CNS), primary CNS lymphoma, tumor angiogenesis, spinal axis tumor, brain stem glioma, pituitary adenoma, Kaposi's sarcoma, epidermoid cancer, squamous cell cancer, T-cell lymphoma, environmentally induced cancers, combinations of said cancers, and metastatic lesions of said cancers; and / or the cancer is a hematologic cancer chosen from one or more of chronic lymphocytic leukemia (CLL), acute leukemias, acute lymphoid leukemia (ALL), B-cell acute lymphoid leukemia (B-ALL), T-cell acute lymphoid leukemia (T-ALL), chronic myelogenous leukemia (CML), B cell prolymphocyticleukemia, blastic plasmacytoid dendritic cell neoplasm, Burkitt's lymphoma, diffuse large B cell lymphoma, follicular lymphoma, hairy cell leukemia, small cell- or a large cell-follicular lymphoma, malignant lymphoproliferative conditions, MALT lymphoma, mantle cell lymphoma, marginal zone lymphoma, multiple myeloma, myelodysplasia and myelodysplastic syndrome, non-Hodgkin's lymphoma, Hodgkin's lymphoma, plasmablastic lymphoma, plasmacytoid dendritic cell neoplasm, Waldenstrom macroglobulinemia, or pre-leukemia. In some embodiments, administering comprises aerosol delivery, nasal delivery, vaginal delivery, rectal delivery, buccal delivery, ocular delivery, local delivery, topical delivery, intraci sternal delivery, intraperitoneal delivery, oral delivery, intramuscular injection, intravenous injection, subcutaneous injection, intranodal injection, intratumoral injection, intraperitoneal injection, intradermal injection, or any combination thereof. In some embodiments, the composition is administered intramuscularly. In some embodiments, the composition is administered into a deltoid region of an arm.BRIEF DESCRIPTION OF THE DRAWINGS

[0068] FIG. 1A-FIG. 1C depict exemplary schematics of the mechanisms of action of mRNA vaccines (FIG. 1A), virus-like particle based vaccines (FIG. IB), and the hybrid ENP vaccines of the disclosure (FIG. 1C). mRNA vaccines mimic infected cells to activate B- and T- cells (FIG. 1 A). Virus-Like Particle (VLP)-based vaccines mimic viruses to activate B-cells (FIG. IB). Hybrid mRNA vaccines encoding self-assembling VLPs enhance B-cell activation (FIG. 1C).

[0069] FIG. 2 displays a non-limiting exemplary schematic depicting EABR domain recruitment of ESCRT proteins to viral spike proteins to induce VLP assembly.

[0070] FIG. 3 displays a non-limiting exemplary schematic of the domain structure of a fusion protein of the disclosure.

[0071] FIG. 4 depicts non-limiting exemplary data showing Spike-EABR mRNA vaccine elicits >5-fold higher neutralizing antibody titers. In the graph, for each time point from left to right is shown: S-EABR VLPs + adjuvant, S mRNA, and S-EABR mRNA.

[0072] FIG. 5A-FIG. 5B display an exemplary schematic presentation comparing the original ERD approach and the ERD adapter system. FIG. 5A displays the previously described ERD approach, in which ERDs are fused directly to the cytoplasmic domain of the membranebound immunogen. ERD-mediated recruitment of ESCRT proteins induces ENP budding. An exemplary diagram of an ERD adapter system of the present disclosure is shown in FIG. 5B. The ERD is directly fused to a small adapter protein that targets the ERD to the plasma membrane. The adapter protein specifically interacts with the modified cytoplasmic domain of the immunogen, thereby connecting the ERD to the immunogen. In some embodiments, ERD adapter designs take advantage of the known interaction between the cytoplasmic domain of the humanCD4 protein and the N-terminal unique domain of the human tyrosine kinase Lek involved in T cell activation and signaling. The cytoplasmic domain of the SARS-CoV-2 spike protein was replaced with the human CD4 cytoplasmic tail (CD4 CT). The ERD adapter was designed by fusing residues 1-61 of Lek, containing the SH4 and unique domains, to the ERD separated by a 12-residue flexible linker. The Lck-based ERD adapter localizes to the plasma membrane due to myristoylation of the glycine residue at position 2 and palmitoylation of the cysteine residues at positions 3 and 5. Targeting the ERD to the plasma membrane promotes efficient ENP budding and facilitates interactions with the membrane-bound immunogen leading to its incorporation into the budding ENP.

[0073] FIG. 6A-FIG. 6B display non-limiting exemplary data showing the ERD adapter system induces efficient budding of ENPs displaying SARS-CoV-2 spike. mRNAs encoding SARS-CoV-2 spike, spike-EPM-EABR, spike-CD4 CT, spike-CD4 CT-EPM, spike- CD4 CT (1.5 pg) + Lck-EABR (0.5 pg), spike-CD4 CT (1.8 pg) + Lck-EABR (0.2 pg), spike- CD4 CT-EPM (1.5 pg) + Lck-EABR (0.5 pg), or spike-CD4 CT-EPM (1.8 pg) + Lck-EABR (0.2 pg) were synthesized and transfected into HEK293T cells. 48 hours post-transfection, cell surface expression of spike was analyzed by flow cytometry (FIG. 6A). To quantify ENP budding, transfected cell culture supernatants were harvested and ENPs were purified by ultracentrifugation on a 20% sucrose cushion. Purified ENP samples were serially diluted, and spike protein levels were measured by ELISA (FIG. 6B).

[0074] FIG. 7A-FIG. 7B display non-limiting exemplary data showing the ERD adapter system induces efficient budding of ENPs displaying influenza HA. mRNAs encoding influenza HA, HA-EPM-EABR, HA-CD4 CT, HA-CD4 CT + Lek, HA-CD4 CT + Lck-EABR, or HA-muCD4 CT + muLck-EABR were synthesized and transfected into HEK293T cells. 48 hours post-transfection, cell surface expression of HA was analyzed by flow cytometry (FIG. 7A). To quantify ENP budding, transfected cell culture supernatants were harvested and ENPs were purified by ultracentrifugation on a 20% sucrose cushion. Purified ENP samples were serially diluted, and HA protein levels were measured by ELISA (FIG. 7B).

[0075] FIG. 8A-FIG. 8B display non-limiting exemplary data showing that removal of dileucine endocytosis motif from CD4 CT improves cell surface expression and ENP budding. mRNAs encoding SARS-CoV-2 spike + Lck-EABR, spike-CD4 CT + Lck-EABR, spike-CD4 CT-delEM + Lck-EABR, spike-CD4 CT-delEMl + Lck-EABR, or spike-CD4 CT-delEM2 + Lck- EABR were synthesized and transfected into HEK293T cells. 48 hours post-transfection, cell surface expression of spike was analyzed by flow cytometry (FIG. 8 A). To quantify ENP budding, transfected cell culture supernatants were harvested and ENPs were purified by ultracentrifugation on a 20% sucrose cushion. Purified ENP samples were serially diluted, and spike protein levelswere measured by ELISA (FIG. 8B).

[0076] FIG. 9A-FIG. 9B display non-limiting exemplary data showing that removal of dileucine endocytosis motif from muCD4 CT does not improve ENP budding. mRNAs encoding SARS-CoV-2 spike-CD4 CT-delEMl + Lck-EABR, spike-muCD4 CT + muLck- EABR, spike-muCD4 CT-delEM + muLck-EABR, spike-muCD4 CT-delEMl + muLck-EABR, or spike-muCD4 CT-delEM2 + muLck-EABR were synthesized and transfected into HEK293T cells. 48 hours post-transfection, cell surface expression of spike was analyzed by flow cytometry (FIG. 9 A). To quantify ENP budding, transfected cell culture supernatants were harvested and ENPs were purified by ultracentrifugation on a 20% sucrose cushion. Purified ENP samples were serially diluted, and spike protein levels were measured by ELISA (FIG. 9B).

[0077] FIG. 10A-FIG. 10B display non-limiting exemplary data showing that the Lck- CD4 CT-delEM interaction promotes more efficient ENP budding than the Lck-CD8 CT interaction. mRNAs encoding SARS-CoV-2 spike-CD4 CT-delEM + Lck-EABR or spike-CD8 CT + Lck-EABR were synthesized and transfected into HEK293T cells. 48 hours posttransfection, cell surface expression of spike was analyzed by flow cytometry (FIG. 10A). To quantify ENP budding, transfected cell culture supernatants were harvested and ENPs were purified by ultracentrifugation on a 20% sucrose cushion. Purified ENP samples were serially diluted, and spike protein levels were measured by ELISA (FIG. 10B).

[0078] FIG. 11 A-FIG. 1 IB display non-limiting exemplary data showing introduction of disulfide bonds between CD4 CT and Lek improves ENP budding. mRNAs encoding SARS- CoV-2 spike-CD4 CT + Lck-EABR, spike-CD4 CT-A404C + Lck-E15C-EABR, spike-CD4 CT- 1410C + Lck-M14C-EABR, or spike-CD4 CT-T419C + Lck-P26C-EABR were synthesized and transfected into HEK293T cells. 48 hours post-transfection, cell surface expression of spike was analyzed by flow cytometry (FIG. 11 A). To quantify ENP budding, transfected cell culture supernatants were harvested and ENPs were purified by ultracentrifugation on a 20% sucrose cushion. Purified ENP samples were serially diluted, and spike protein levels were measured by ELISA (FIG. 1 IB).

[0079] FIG. 12A-FIG. 12B display non-limiting exemplary data showing introduction of disulfide bond between CD4 CT-delEMl and Lek does not improve ENP budding. mRNAs encoding SARS-CoV-2 spike, spike-CD4 CT-delEMl + Lck-EABR, or spike-CD4 CT-delEMl- A404C + Lck-E15C-EABR were synthesized and transfected into HEK293T cells. 48 hours posttransfection, cell surface expression of spike was analyzed by flow cytometry (FIG. 12A). To quantify ENP budding, transfected cell culture supernatants were harvested and ENPs were purified by ultracentrifugation on a 20% sucrose cushion. Purified ENP samples were serially diluted, and spike protein levels were measured by ELISA (FIG. 12B).

[0080] FIG. 13A-FIG. 13B display non-limiting exemplary data showing various ERD adapters induce high cell surface expression and efficient ENP budding. mRNAs encoding SARS- CoV-2 spike, spike-EABR, spike-CD4 CT-delEMl + Lck-EABR, spike-CD4 CT-delEMl + Lck- p6v5.1, spike-CD4 CT-delEMl + Lck-rGalectin-3mini-ALIX2, or spike-CD4 CT-delEMl + Lck- EABR + Lck-p6v5.1 were synthesized and transfected into HEK293T cells. 48 hours posttransfection, cell surface expression of spike was analyzed by flow cytometry (FIG. 13A). To quantify ENP budding, transfected cell culture supernatants were harvested and ENPs were purified by ultracentrifugation on a 20% sucrose cushion. Purified ENP samples were serially diluted, and spike protein levels were measured by ELISA (FIG. 13B).

[0081] FIG. 14A-FIG. 14B display non-limiting exemplary data showing ACP33- based ERD adapters do not induce ENP budding. mRNAs encoding SARS-CoV-2 spike-CD4 CT- delEMl + Lck-EABR, spike-CD4 CT-delEMl + ACP33-EABR, spike-CD4 CT-delEMl + Lck- p6v5.1, or spike-CD4 CT-delEMl + ACP33-p6v5.1 were synthesized and transfected into HEK293T cells. 48 hours post-transfection, cell surface expression of spike was analyzed by flow cytometry (FIG. 14A). To quantify ENP budding, transfected cell culture supernatants were harvested and ENPs were purified by ultracentrifugation on a 20% sucrose cushion. Purified ENP samples were serially diluted, and spike protein levels were measured by ELISA (FIG. 14B).

[0082] FIG. 15A-FIG. 15B display non-limiting exemplary data showing coexpression of Lek- and ACP33-based adapters induces small improvements in cell surface expression and / or ENP budding. mRNAs encoding SARS-CoV-2 spike, spike-CD4 CT-delEMl + Lck-EABR, spike-CD4 CT-delEMl + Lck-p6v5.1, spike-CD4 CT-delEMl + Lck-EABR + ACP33-EABR, spike-CD4 CT-delEMl + Lck-EABR + ACP33-p6v5.1, spike-CD4 CT-delEMl + Lck-p6v5.1 + ACP33-EABR, or spike-CD4 CT-delEMl + Lck-p6v5.1 + ACP33-p6v5.1 were synthesized and transfected into HEK293T cells. 48 hours post-transfection, cell surface expression of spike was analyzed by flow cytometry (FIG. 15 A). To quantify ENP budding, transfected cell culture supernatants were harvested and ENPs were purified by ultracentrifugation on a 20% sucrose cushion. Purified ENP samples were serially diluted, and spike protein levels were measured by ELISA (FIG. 15B).

[0083] FIG. 16A-FIG. 16B display non-limiting exemplary data showing coexpression of HA-CD4 CT-delEMl .2 and Lck-p6v5.1 induces high cell surface expression and ENP budding. mRNAs encoding influenza HA, HA-EPM-EABR, HA-CD4 CT-delEMl .1 + Lck- p6v5.1, or HA-CD4 CT-delEMl .2 + Lck-p6v5.1 were synthesized and transfected into HEK293T cells. 48 hours post-transfection, cell surface expression of HA was analyzed by flow cytometry (FIG. 16A). To quantify ENP budding, transfected cell culture supernatants were harvested and ENPs were purified by ultracentrifugation on a 20% sucrose cushion. Purified ENP samples wereserially diluted, and HA protein levels were measured by ELISA (FIG. 16B).

[0084] FIG. 17A-FIG. 17B display non-limiting exemplary data showing insertion of EPM into Lck-EABR adapter slightly increases ENP budding. mRNAs encoding SARS-CoV-2 spike, spike-CD4 CT-delEMl + Lck-EABR, spike-CD4 CT-delEMl + Lck-p6v5.1, spike-CD4 CT-delEMl + Lck-EPM-EABR, or spike-CD4 CT-delEMl + Lck-EPM-p6v5.1 were synthesized and transfected into HEK293T cells. 48 hours post-transfection, cell surface expression of spike was analyzed by flow cytometry (FIG. 17A). To quantify ENP budding, transfected cell culture supernatants were harvested and ENPs were purified by ultracentrifugation on a 20% sucrose cushion. Purified ENP samples were serially diluted, and spike protein levels were measured by ELISA (FIG. 17B).

[0085] FIG. 18A-FIG. 18B display non-limiting exemplary data showing Lck-p6v6 adapters increase ENP budding for SARS-CoV-2 spike immunogen. mRNAs encoding SARS- CoV-2 spike, spike-EPM-EABR, spike-CD4 CT-delEMl + Lck-p6v5.1, spike-CD4 CT-delEMl + Lck-p6v6.1, spike-CD4 CT-delEMl + Lck-p6v6.2, spike-CD4 CT-delEMl + Lck-p6v6.3, spike-CD4 CT-delEMl + Lck-p6v6.4, spike-CD4 CT-delEMl + Lck-p6v6.5, spike-CD4 CT- delEMl + Lck-p6v6.6, spike-CD4 CT-delEMl + Lck-p6v6.7, spike-CD4 CT-delEMl + Lck- p6v6.8, spike-CD4 CT-delEMl + Lck-p6v6.9, spike-CD4 CT-delEMl + Lck-p6v6.10 (rose), or spike-CD4 CT-delEMl + Lck-p6v6.11 were synthesized and transfected into HEK293T cells. 48 hours post-transfection, cell surface expression of spike was analyzed by flow cytometry (FIG. 18A). To quantify ENP budding, transfected cell culture supernatants were harvested and ENPs were purified by ultracentrifugation on a 20% sucrose cushion. Purified ENP samples were serially diluted, and spike protein levels were measured by ELISA (FIG. 18B).

[0086] FIG. 19A-FIG. 19B display non-limiting exemplary data showing Lck-p6v6 adapters increase ENP budding for influenza HA immunogen. mRNAs encoding influenza HA, HA-EPM-EABR, HA-CD4 CT-delEMl.2 + Lck-p6v5.1, HA-CD4 CT-delEMl.2 + Lck-p6v6.1, HA-CD4 CT-delEMl.2 + Lck-p6v6.2, HA-CD4 CT-delEMl.2 + Lck-p6v6.3, HA-CD4 CT- delEM1.2 + Lck-p6v6.4, HA-CD4 CT-delEMl.2 + Lck-p6v6.5, HA-CD4 CT-delEMl.2 + Lck- p6v6.6, HA-CD4 CT-delEMl.2 + Lck-p6v6.7, HA-CD4 CT-delEMl.2 + Lck-p6v6.8, HA-CD4 CT-delEMl.2 + Lck-p6v6.9, HA-CD4 CT-delEMl.2 + Lck-p6v6.10, or HA-CD4 CT-delEMl.2 + Lck-p6v6.11 were synthesized and transfected into HEK293T cells. 48 hours post-transfection, cell surface expression of HA was analyzed by flow cytometry (FIG. 19A). To quantify ENP budding, transfected cell culture supernatants were harvested and ENPs were purified by ultracentrifugation on a 20% sucrose cushion. Purified ENP samples were serially diluted, and HA protein levels were measured by ELISA (FIG. 19B).

[0087] FIG. 20A-FIG. 20B display non-limiting exemplary data showing Lck-CD4CT sequences from other species induce lower ENP budding. mRNAs encoding SARS-CoV-2 spike-CD4 CT-delEMl + Lck-EABR, spike-chickCD4 CT-delEMl + chickLck-EABR, spike- dragCD4 CT-delEMl + dragLck-EABR, or spike-snakeCD4 CT-delEMl + snakeLck-EABR were synthesized and transfected into HEK293T cells. 48 hours post-transfection, cell surface expression of spike was analyzed by flow cytometry (FIG. 20 A). To quantify ENP budding, transfected cell culture supernatants were harvested and ENPs were purified by ultracentrifugation on a 20% sucrose cushion. Purified ENP samples were serially diluted, and spike protein levels were measured by ELISA (FIG. 20B).

[0088] FIG. 21A-FIG. 21B display non-limiting exemplary data showing ERD adapters systems using heterodimeric coiled-coil interactions promote efficient ENP budding. mRNAs encoding influenza HA, HA-CD4 CT-delEMl.2 + Lck-EABR, HA-ACIDpl + Fyn- BASEpl-EABR, HA-BASEpl + Fyn-ACIDpl-EABR, or HA-N5 + Fyn-N6-EABR were synthesized and transfected into HEK293T cells. 48 hours post-transfection, cell surface expression of HA was analyzed by flow cytometry (FIG. 21A). To quantify ENP budding, transfected cell culture supernatants were harvested and ENPs were purified by ultracentrifugation on a 20% sucrose cushion. Purified ENP samples were serially diluted, and HA protein levels were measured by ELISA (FIG. 2 IB).

[0089] FIG. 22A-FIG. 22C display a non-limiting exemplary schematic presentation of ENPs that co-display the immunogen and an immune cell-targeting protein. The previously described ERD approach is shown in FIG. 22A. An ERD such as the EABR is fused directly to the cytoplasmic domain of the membrane-bound immunogen. ERD-mediated recruitment of ESCRT proteins induces ENP budding. FIG. 22B-FIG. 22C show non-limiting exemplary schematics of co-localization of a membrane-bound immunogen and, e.g., an immune celltargeting protein on the cell surface results in co-di splay of both proteins on the ENP surface. The ERD can be fused to the cytoplasmic domain of (FIG. 22B) the immunogen or (FIG. 22C) the immune cell-targeting protein.

[0090] FIG. 23 displays non-limiting exemplary data showing co-expression of SARS-CoV-2 spike and muC3d-EPM-EABR produces ENPs that display high levels of spike. DNA plasmids encoding SARS-CoV-2 spike-EPM-EABR, spike-EPM-EABR + muC3d, spike + muC3d-EPM-EABR, or spike-EPM-EABR + muC3d-EPM-EABR were transfected into Expi293 cells. 72 hours post-transfection, cell culture supernatants were harvested and ENPs were purified by ultracentrifugation on a 20% sucrose cushion. Purified ENP samples were serially diluted, and spike protein levels were measured by ELISA.

[0091] FIG. 24 displays non-limiting exemplary data showing ENPs co-displaying SARS-CoV-2 spike and muC3d can be targeted to muCR2-expressing cells. HEK293T cells weretransiently transfected to express muCR2. The cells were mixed with the purified ENP samples from FIG. 23 at a concentration of 5 pg / mL (quantified based on spike protein levels in purified ENP samples) for 30 min. The cells were then stained with an anti-spike antibody and a fluorescently-labeled secondary antibody, and samples were analyzed by flow cytometry. For this experimental design, muCR2-expressing cells can only be fluorescently-labeled when they bind to ENPs that co-display muC3d and spike to interact with muCR2 and the anti-spike antibody, respectively. Purified ENP samples included spike-EPM-EABR, spike-EPM-EABR + muC3d, spike + muC3d-EPM-EABR, or spike-EPM-EABR + muC3d-EPM-EABR.

[0092] FIG. 25A-FIG. 25B display non-limiting exemplary data showing co-delivery of mRNAs encoding SARS-CoV-2 spike and muC3d-EPM-EABR promotes high levels of spike immunogen on the cell surface and ENPs. mRNAs encoding SARS-CoV-2 spike, spike-EPM- EABR, spike + muC3d-EPM-EABR, or spike + flagellintrunc-EPM-EABR were synthesized and transfected into HEK293T cells. 48 hours post-transfection, cell surface expression of spike was analyzed by flow cytometry (FIG. 25A). To quantify ENP budding, transfected cell culture supernatants were harvested and ENPs were purified by ultracentrifugation on a 20% sucrose cushion. Purified ENP samples were serially diluted, and spike protein levels were measured by ELISA (FIG. 25B).

[0093] FIG. 26A-FIG. 26D display non-limiting exemplary data showing tetherin expression blocks ENP budding for SARS-CoV-2 spike-EPM-EABR and influenza HA-EPM- EABR fusion constructs. In FIG. 26A-FIG. 26B, mRNAs encoding SARS-CoV-2 spike and spike- EPM-EABR were synthesized and transfected into HEK293T cells. Spike-EPM-EABR was also co-transfected with indicated amounts of mRNA-encoded tetherin. 48 hours post-transfection, cell surface expression of spike was analyzed by flow cytometry (FIG 26A). To quantify ENP budding, transfected cell culture supernatants were harvested and ENPs were purified by ultracentrifugation on a 20% sucrose cushion. Purified ENP samples were serially diluted, and spike protein levels were measured by ELISA (FIG. 26B). In FIG. 26C-FIG. 26D, mRNAs encoding influenza HA and HA-EPM-EABR were synthesized and transfected into HEK293T cells. HA-EPM-EABR was also co-transfected with indicated amounts of mRNA-encoded tetherin. 48 hours post-transfection, cell surface expression of HA was analyzed by flow cytometry (FIG. 26C). To quantify ENP budding, transfected cell culture supernatants were harvested and ENPs were purified by ultracentrifugation on a 20% sucrose cushion. Purified ENP samples were serially diluted, and HA protein levels were measured by ELISA (FIG. 26D).

[0094] FIG. 27A-FIG. 27B display non-limiting exemplary data showing the p6v4.2 ERD is less sensitive to tetherin-mediated blockage of ENP budding compared to other ERDs. mRNAs encoding influenza HA, HA-EPM-EABR, HA-p6v4.2, HA-EPM-rGalectin-3mini, or HA-CD4 CT-delEM1.2 + Lck-EABR were synthesized and transfected into HEK293T cells. HA- EPM-EABR, HA-p6v4.2, HA-EPM-rGalectin-3mini, or HA-CD4 CT-delEM1.2 + Lck-EABR were also co-transfected with 0.025 pg of mRNA-encoded tetherin. 48 hours post-transfection, cell surface expression of HA was analyzed by flow cytometry (FIG. 27A). To quantify ENP budding, transfected cell culture supernatants were harvested and ENPs were purified by ultracentrifugation on a 20% sucrose cushion. Purified ENP samples were serially diluted, and HA protein levels were measured by ELISA (FIG. 27B).

[0095] FIG. 28A-FIG. 28D display non-limiting exemplary data showing HIV-1 Vpu co-expression antagonizes tetherin-mediated blockage of ENP budding. In FIG. 28A-FIG. 28B, mRNAs encoding SARS-CoV-2 spike and spike-EPM-EABR were synthesized and transfected into HEK293T cells. Spike-EPM-EABR was also co-transfected with 0.025 pg of mRNA- encoded tetherin, 0.05 pg of mRNA-encoded Vpu, 0.025 pg of mRNA-encoded tetherin and 0.05 pg of mRNA-encoded Vpu, 0.025 pg of mRNA-encoded Vpu, or 0.025 pg of mRNA-encoded tetherin and 0.025 pg of mRNA-encoded Vpu. 48 hours post-transfection, cell surface expression of spike was analyzed by flow cytometry (FIG. 28A). To quantify ENP budding, transfected cell culture supernatants were harvested and ENPs were purified by ultracentrifugation on a 20% sucrose cushion. Purified ENP samples were serially diluted, and spike protein levels were measured by ELISA (FIG. 28B). In FIG. 28C-FIG. 28D, mRNAs encoding influenza HA and HA- EPM-EABR were synthesized and transfected into HEK293T cells. HA-EPM-EABR was also cotransfected with 0.025 pg of mRNA-encoded tetherin, 0.05 pg of mRNA-encoded Vpu, 0.025 pg of mRNA-encoded tetherin and 0.05 pg of mRNA-encoded Vpu, 0.025 pg of mRNA-encoded Vpu, or 0.025 pg of mRNA-encoded tetherin and 0.025 pg of mRNA-encoded Vpu. 48 hours post-transfection, cell surface expression of HA was analyzed by flow cytometry (FIG. 28C). To quantify ENP budding, transfected cell culture supernatants were harvested and ENPs were purified by ultracentrifugation on a 20% sucrose cushion. Purified ENP samples were serially diluted, and HA protein levels were measured by ELISA (FIG. 28D).

[0096] FIG. 29A-FIG. 29B display non-limiting exemplary data showing KSHV K5 and SARS-CoV-2 ORF7a do not antagonize tetherin-mediated blockage of ENP budding. mRNAs encoding influenza HA and HA-EPM-EABR were synthesized and transfected into HEK293T cells. HA-EPM-EABR was also co-transfected with 0.025 pg of mRNA-encoded tetherin, 0.025 pg of mRNA-encoded tetherin and 0.1 pg of mRNA-encoded Vpu, 0.025 pg of mRNA-encoded tetherin and 0.025 pg of mRNA-encoded Vpu, 0.025 pg of mRNA-encoded tetherin and 0.1 pg of mRNA-encoded K5, 0.025 pg of mRNA-encoded tetherin and 0.025 pg of mRNA-encoded K5, 0.025 pg of mRNA-encoded tetherin and 0.1 pg of mRNA-encoded ORF7a, 0.025 pg of mRNA-encoded tetherin and 0.025 pg of mRNA-encoded ORF7a. 48 hours post-transfection, cellsurface expression of HA was analyzed by flow cytometry (FIG. 29A). To quantify ENP budding, transfected cell culture supernatants were harvested and ENPs were purified by ultracentrifugation on a 20% sucrose cushion. Purified ENP samples were serially diluted, and HA protein levels were measured by ELISA (FIG. 29B).DETAILED DESCRIPTION

[0097] In the following detailed description, reference is made to the accompanying drawings, which form a part hereof. In the drawings, similar symbols typically identify similar components, unless context dictates otherwise. The illustrative embodiments described in the detailed description, drawings, and claims are not meant to be limiting. Other embodiments may be utilized, and other changes may be made, without departing from the spirit or scope of the subject matter presented herein. It will be readily understood that the aspects of the present disclosure, as generally described herein, and illustrated in the Figures, can be arranged, substituted, combined, separated, and designed in a wide variety of different configurations, all of which are explicitly contemplated herein and made part of the disclosure herein.

[0098] All patents, published patent applications, other publications, and sequences from GenBank, and other databases referred to herein are incorporated by reference in their entirety with respect to the related technology.

[0099] Disclosed herein include compositions. In some embodiments, the composition comprises: a nucleic acid composition comprising: (i) a polynucleotide encoding a dimerization fusion protein, wherein the dimerization fusion protein comprises an optional antigenic polypeptide (AP) and a heterologous cytoplasmic tail, and (ii) a polynucleotide encoding an adapter fusion protein comprising an endosomal sorting complex required for transport (ESCRT)- recruiting domain (ERD) and an adapter domain capable of binding the heterologous cytoplasmic tail to form a heterodimer, wherein binding of the adapter domain to the heterologous cytoplasmic tail is capable of recruiting one or more ESCRT proteins to the heterodimer, thereby inducing a plurality of dimerization fusion proteins to self-assemble into an enveloped nanoparticle (ENP) secreted from a cell in which the dimerization fusion protein and adapter fusion protein are expressed, thereby generating a population of ENPs.

[0100] In some embodiments, the composition comprises: a nucleic acid composition comprising (i) n polynucleotides each encoding an nth dimerization fusion protein, wherein n is an integer from 2 to 500, wherein each dimerization fusion protein comprises an optional antigenic polypeptide (AP) and a heterologous cytoplasmic tail, wherein at least two of the dimerization fusion proteins differ with respect to the AP; and (ii) a polynucleotide encoding an adapter fusion protein comprising an endosomal sorting complex required for transport (ESCRT)-recruiting domain (ERD) and an adapter domain capable of binding the heterologous cytoplasmic tail toform a heterodimer, wherein binding of the adapter domain to the heterologous cytoplasmic tail is capable of recruiting one or more ESCRT proteins to the heterodimer, thereby inducing a plurality of dimerization fusion proteins to self-assemble into an enveloped nanoparticle (ENP) secreted from a cell in which the n dimerization fusion proteins and the adapter fusion protein are expressed, thereby generating a population of ENPs.

[0101] In some embodiments, the composition comprises: a population of enveloped nanoparticles (ENPs), wherein each of the ENPs comprises a plurality of the dimerization fusion proteins each comprising a heterologous cytoplasmic tail and optionally, an antigenic polypeptide (AP).

[0102] In some embodiments, the composition comprises: a nucleic acid composition comprising: (i) a polynucleotide encoding a recombinant protein comprising an optional antigenic polypeptide (AP) and a transmembrane domain, and (ii) a polynucleotide encoding a chimeric protein comprising: a) a signal peptide, b) a cell surface domain, c) a transmembrane domain, and d) a cytoplasmic domain; wherein the recombinant protein and / or the chimeric protein further comprises an endosomal sorting complex required for transport (ESCRT)-recruiting domain (ERD), optionally the recombinant protein further comprises a cytoplasmic domain; and wherein the recombinant protein and the chimeric protein are capable of co-localizing to a site on the plasma membrane of the cell, and wherein the ERD is capable of recruiting one or more ESCRT proteins to the site on the plasma membrane, thereby inducing a plurality of recombinant proteins and chimeric proteins to self-assemble into an enveloped nanoparticle (ENP) secreted from a cell in which the recombinant protein and chimeric protein are expressed, thereby generating a population of ENPs, optionally the cytoplasmic domain of the recombinant protein and the cytoplasmic domain of the chimeric protein are capable of co-localizing the recombinant protein and the chimeric protein to a site on the plasma membrane of the cell, optionally the transmembrane domain of the recombinant protein and the transmembrane domain of the chimeric protein are capable of co-localizing the recombinant protein and the chimeric protein to a site on the plasma membrane of the cell.

[0103] Disclosed herein include compositions. In some embodiments, the composition comprises: a nucleic acid composition comprising: (i) n polynucleotides each encoding a recombinant protein comprising an optional antigenic polypeptide (AP) and a transmembrane domain, wherein at least two of the recombinant proteins differ with respect to the AP, wherein n is an integer from 2 to 500; and (ii) z polynucleotides each encoding a chimeric protein comprising: a) a signal peptide, b) a cell surface domain, wherein at least two of the chimeric proteins differ with respect to the cell surface domain, c) a transmembrane domain, and d) a cytoplasmic domain wherein z is an integer from 1 to 500, wherein the nth recombinant proteinsand / or the zth chimeric proteins further comprise an endosomal sorting complex required for transport (ESCRT)-recruiting domain (ERD), optionally the nth recombinant proteins further comprise a cytoplasmic domain, and wherein each of the nth recombinant protein and each of the zth chimeric protein are capable of co-localizing to a site on the plasma membrane of the cell, and the ERD is capable of recruiting one or more ESCRT proteins to the site on the plasma membrane, thereby inducing a plurality of recombinant proteins and chimeric proteins to self-assemble into an enveloped nanoparticle (ENP) secreted from the cell in which the recombinant proteins and chimeric proteins are expressed, thereby generating a population of ENPs, optionally the cytoplasmic domain of each of the nth recombinant protein and the cytoplasmic domain of each of the zth chimeric protein are capable of co-localizing each of the nth recombinant protein and each of the zth chimeric protein to a site on the plasma membrane of the cell, optionally the transmembrane domain of each of the nth recombinant protein and the transmembrane domain of each of the zth chimeric protein are capable of co-localizing each of the nth recombinant protein and each of the zth chimeric protein to a site on the plasma membrane of the cell.

[0104] Disclosed herein include kits. In some embodiments, the kit comprises: a composition disclosed herein (e.g., a nucleic acid composition, a population of ENPs). Disclosed herein include cells. In some embodiments, a cell comprises: a nucleic acid composition disclosed herein.

[0105] Disclosed herein include methods of stimulating an immune response in a subject in need thereof. In some embodiments, the method comprises: administering to the subject a pharmaceutically effective amount of a composition disclosed herein (e.g., a nucleic acid composition, a population of ENPs), thereby stimulating an immune response in the subject.

[0106] Disclosed herein include methods of treating or preventing a disease or disorder in a subject in need thereof. In some embodiments, the method comprises: administering to the subject a pharmaceutically effective amount of a composition disclosed herein (e.g., a nucleic acid composition, a population of ENPs), thereby treating or preventing the disease or disorder in the subject. In some embodiments, the disease or disorder is a disease or disorder caused by an infectious agent. In some embodiments, the disease or disorder caused by an infectious agent is a disease or disorder caused by a coronavirus (CoV) infection.

[0107] Disclosed herein include methods for treating or preventing a coronavirus (CoV) infection in a subject in need thereof. In some embodiments, the method comprises: administering to the subject a pharmaceutically effective amount of a composition disclosed herein (e.g., a nucleic acid composition, a population of ENPs), thereby treating or preventing the CoV infection in the subject.Definitions

[0108] Unless defined otherwise, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the present disclosure belongs. See, e.g. Singleton et al., Dictionary of Microbiology and Molecular Biology 2nd ed., J. Wiley & Sons (New York, NY 1994); Sambrook et al., Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Press (Cold Spring Harbor, NY 1989). For purposes of the present disclosure, the following terms are defined below.

[0109] As used herein, the terms “antigen” or “immunogen” are used interchangeably to refer to a substance, typically a protein, which is capable of inducing an immune response in a subject (e.g., a mammal, such as a human). The term also refers to proteins that are immunologically active in the sense that once administered to a subject, either directly or in the form of a nucleotide sequence or vector that encodes the protein, is able to evoke an immune response of the humoral and / or cellular type directed against that protein or a variant thereof.

[0110] As used herein, “sequence identity” or “identity” in the context of two nucleic acid or polypeptide sequences makes reference to the nucleotide bases or residues in the two sequences that are the same when aligned for maximum correspondence over a specified comparison window. Methods of alignment of sequences for comparison are well known in the art. Various programs and alignment algorithms are described in: Smith & Waterman, Adv. Appl. Math. 2:482, 1981; Needleman & Wunsch, J. Mol. Biol. 48:443, 1970; Pearson & Lipman, Proc. Natl. Acad. Sci. USA 85:2444, 1988; Higgins & Sharp, Gene, 73:237-44, 1988; Higgins & Sharp, CABIOS 5: 151-3, 1989; Corpet et al., Nuc. Acids Res. 16: 10881-90, 1988; Huang et al. Computer Appls. in the Biosciences 8, 155-65, 1992; Pearson et al., Meth. Mol. Bio. 24:307-31, 1994; and Altschul et al., J. Mol. Biol. 215:403-10, 1990 (the content of each of these references is incorporated herein in its entirety).[OHl] When percentage of sequence identity or similarity is used in reference to proteins, it is recognized that residue positions which are not identical often differ by conservative amino acid substitutions, where amino acid residues are substituted with a functionally equivalent residue of the amino acid residues with similar physiochemical properties and therefore do not change the functional properties of the molecule. A functionally equivalent residue of an amino acid used herein typically can refer to other amino acid residues having physiochemical and stereochemical characteristics substantially similar to the original amino acid. The physiochemical properties include water solubility (hydrophobicity or hydrophilicity), dielectric and electrochemical properties, physiological pH, partial charge of side chains (positive, negative or neutral) and other properties identifiable to one of skill in the art. The stereochemical characteristics include spatial and conformational arrangement of the amino acids and theirchirality. For example, glutamic acid is considered to be a functionally equivalent residue to aspartic acid in the sense of the current disclosure. Tyrosine and tryptophan are considered as functionally equivalent residues to phenylalanine. Arginine and lysine are considered as functionally equivalent residues to histidine.

[0112] The term “substantially identical” as used herein in the context of two or more sequences refers to a specified percentage of amino acid residues or nucleotides that are identical or functionally equivalent, such as about, at least or at least about 65% identity, optionally, about, at least or at least about 70%, 75%, 80%, 85%, 90%, 95%, or 99% identity over a specified region or over the entire sequence.

[0113] As used herein, the term “variant” refers to a polynucleotide or polypeptide having a sequence substantially similar or identical to a reference (e.g., the parent) polynucleotide or polypeptide. In the case of a polynucleotide, a variant can have deletions, substitutions, additions of one or more nucleotides at the 5' end, 3' end, and / or one or more internal sites in comparison to the reference polynucleotide. Similarities and / or differences in sequences between a variant and the reference polynucleotide can be detected using conventional techniques known in the art, for example polymerase chain reaction (PCR) and hybridization techniques. Variant polynucleotides also include synthetically derived polynucleotides, such as those generated, for example, by using site-directed mutagenesis. Generally, a variant of a polynucleotide, including, but not limited to, a DNA, can have at least, or at least about, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to the reference polynucleotide as determined by sequence alignment programs known in the art. In the case of a polypeptide, a variant can have deletions, substitutions, additions of one or more amino acids in comparison to the reference polypeptide. Similarities and / or differences in sequences between a variant and the reference polypeptide can be detected using conventional techniques known in the art, for example Western blot. A variant of a polypeptide can have, for example, at least, or at least about, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to the reference polypeptide as determined by sequence alignment programs known in the art.

[0114] Standard techniques can be used for recombinant DNA, oligonucleotide synthesis, and tissue culture and transformation (e.g., electroporation, lipofection). Enzymatic reactions and purification techniques can be performed according to manufacturer's specifications or as commonly accomplished in the art or as described herein. The foregoing techniques and procedures can be generally performed according to conventional methods well known in the art and as described in various general and more specific references that are cited and discussed throughout the present specification. See, e.g., Sambrook et al., Molecular Cloning: A LaboratoryManual (2d ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (1989)), which is incorporated herein by reference for any purpose. Unless specific definitions are provided, the nomenclatures utilized in connection with, and the laboratory procedures and techniques of, analytical chemistry, synthetic organic chemistry, and medicinal and pharmaceutical chemistry described herein are those commonly known and used in the art. Standard techniques can be used for chemical syntheses, chemical analyses, pharmaceutical preparation, formulation, and delivery, and treatment of patients.

[0115] The term “construct,” as used herein, refers to a recombinant nucleic acid that has been generated for the purpose of the expression of a specific nucleotide sequence(s), or that is to be used in the construction of other recombinant nucleotide sequences.

[0116] As used herein, the terms “nucleic acid” and “polynucleotide” are interchangeable and refer to any nucleic acid, whether composed of phosphodiester linkages or modified linkages such as phosphotriester, phosphoramidate, siloxane, carbonate, carboxymethylester, acetamidate, carbamate, thioether, bridged phosphoramidate, bridged methylene phosphonate, bridged phosphoramidate, bridged phosphoramidate, bridged methylene phosphonate, phosphorothioate, methylphosphonate, phosphorodithioate, bridged phosphorothioate or sultone linkages, and combinations of such linkages. The terms “nucleic acid” and “polynucleotide” also specifically include nucleic acids composed of bases other than the five biologically occurring bases (adenine, guanine, thymine, cytosine and uracil).

[0117] Hybrid mRNA vaccine approaches utilize genetically encoded self-assembling enveloped nanoparticles (ENPs). Described herein are compositions for improved hybrid vaccines. Described herein are ERD adapter systems, which improve cell surface expression and ENP budding compared to immunogen-ERD fusion constructs. In some embodiments, for this adapter system, the ERD was directly fused to a small adapter protein domain that targets the ERD to the plasma membrane and specifically interacts with the immunogen’s cytoplasmic domain that was modified to tightly bind the adapter, thereby connecting the ERD to the immunogen (FIG. 5A-FIG. 5B). As described herein, this ERD adapter system effectively induces ENP budding for mRNA-encoded SARS-CoV-2 spike and influenza hemagglutinin (HA) immunogens.

[0118] The ERD adapter system of the disclosure was developed because direct fusion of the EABR sequence to the cytoplasmic domain of membrane-bound immunogens can result in low cell surface expression when the mRNA-encoded fusion constructs are delivered to cells. As also described herein, high cell surface expression of the immunogen and efficient ENP budding can be achieved by fusing the EABR to the cytoplasmic domain of a second cell surface protein (e.g., a cell surface protein domain) that co-localizes with the immunogen on the cell surface (FIG. 22A-FIG. 22C). Due to their co-localization, co-delivery of both cell surface proteins can resultin efficient incorporation of the immunogen on the surface of budding ENPs. As the EABR is fused to the second cell surface protein rather than the immunogen, the cell surface expression of the immunogen can be higher compared to immunogen-EABR fusion constructs.

[0119] Without being bound by any particular theory, the second cell surface protein (e.g., a chimeric protein) can further enhance the effectiveness of ERD-based hybrid mRNA vaccines in various ways. First, the second cell surface protein can be a second target antigen for vaccines against pathogens that have multiple surface antigens such as influenza (HA and neuraminidase). In this case, the ERD can be fused to the surface antigen for which it is less critical to achieve high cell surface expression, or to the antigen for which cell surface expression is less affected by EABR fusion to the cytoplasmic domain. The second cell surface protein (e.g., a chimeric protein) can also be a subunit of the immunogen to potentially help focus immune responses to a certain part of the immunogen, e.g., the receptor-binding domain (RBD) of the SARS-CoV-2 spike protein, which is the target of most neutralizing antibodies against SARS- CoV-2. Finally, the second cell surface protein (e.g., a chimeric protein) can be a ligand for cell surface receptors specifically presented on immune cells (FIG. 22A-FIG. 22C). Targeting ENPs to immune cells through co-display of such a targeting protein may enhance immune cell activation and the overall effectiveness of ERD-based hybrid mRNA vaccines. For instance, fusion of complement component 3d (C3d) to protein-based immunogens has been shown to enhance immune responses in vivo. C3d targets immunogens to immune cells by binding to complement receptor 2 (CR2), which is primarily expressed on B cells and follicular dendritic cells.

[0120] As provided herein, a membrane-bound version of mouse C3d (muC3d) can be co-displayed with a SARS-CoV-2 spike immunogen on ENPs and targets ENPs to cells expressing mouse CR2 (muCR2). Importantly, co-transfection of mRNAs encoding a SARS-CoV-2 spike immunogen and a muC3d-EPM-EABR construct induces high cell surface expression of spike and efficient production of ENPs that display similar levels of the spike immunogen compared to a spike-EPM-EABR fusion construct. These results show that immune-cell targeting proteins can be co-displayed on ENPs and that higher cell surface expression of the immunogen can be achieved by fusing the EABR sequence to the cytoplasmic domain of the membrane-bound immune cell-targeting protein instead of the immunogen. The combined effects of immune cell targeting of ENPs, and higher cell surface expression of the immunogen have the potential to improve the effectiveness ERD-based hybrid mRNA vaccines.

[0121] Described herein are ERD adapter systems, which improve cell surface expression and ENP budding compared to immunogen-ERD fusion constructs. For this adapter system, the ERD was directly fused to a small adapter protein (e.g., an adapter domain) that targetsthe ERD to the plasma membrane and specifically interacts with the immunogen’s cytoplasmic domain that was modified to tightly bind the adapter, thereby connecting the ERD to the immunogen. As provided herein, ERD adapter systems effectively induce ENP budding for mRNA-encoded SARS-CoV-2 spike and influenza hemagglutinin (HA) immunogens. The ERD adapter approach can be particularly valuable for immunogen / ERD combinations that result in low cell surface expression of the immunogen when the ERD is directly fused to the cytoplasmic domain of the immunogen. In some embodiments, the ERD adapter design takes advantage of the strong interaction between the CD4 cytoplasmic tail and the N-terminal Lek unique domain. Alternative ERD adapter systems based on heterodimeric coiled-coil peptide pairs were also shown to promote efficient ENP budding. Interestingly, Lck-p6v6-based ERD adapters consistently induced markedly higher cell surface expression than the unmodified membranebound immunogens in addition to high ENP budding that was slightly higher compared to immunogen-EABR fusion constructs. The combination of high cell surface expression and ENP budding can vastly improve the immunogenicity of ERD mRNA vaccines, and this approach can be applied to a wide range of immunogens for the development of protein nanoparticle- or nucleic acid-based vaccines.

[0122] As an alternative approach to increase cell surface expression of the immunogen, also provided herein are membrane-bound immune cell-targeting proteins that colocalize with a SARS-CoV-2 spike immunogen on the cell surface. Due to their co-localization, direct ERD fusion to the cytoplasmic domain of the immunogen or the immune cell-targeting protein produces ENPs that co-display high levels of both proteins. ENPs that co-display immune cell-targeting proteins such as mouse complement component 3d (muC3d) or a truncated bacterial flagellin construct may also enhance the effectiveness of ERD-based hybrid mRNA vaccines by targeting mRNA-encoded ENPs to immune cells. Importantly, co-delivery of mRNAs encoding the membrane-bound spike immunogen and a muC3d-EPM-EABR construct induced high cell surface expression of spike and efficient production of ENPs that displayed similar levels of the spike immunogen compared to a spike-EPM-EABR fusion construct. These results show that immune-cell targeting proteins can be co-displayed on ENPs and that higher cell surface expression of the immunogen can be achieved by fusing the EABR sequence to the cytoplasmic domain of the membrane-bound immune cell-targeting protein instead of the immunogen. The combined effects of immune cell targeting of ENPs, and higher cell surface expression of the immunogen have the potential to improve the effectiveness ERD-based hybrid mRNA vaccines. ENP co-display of TLR agonists such as flagellintrunc can have additional benefits due to their adjuvant properties. The previously described ERD was obtained from the ESCRT- and ALIX- binding region (EABR) of the human CEP55 protein. This ERD induces efficient ENP buddingwhen directly fused to the cytoplasmic tail of various cell surface proteins. However, direct fusion of the EABR sequence to the cytoplasmic domain of SARS-CoV-2 spike can result in lower cell surface expression when the mRNA-encoded fusion constructs are delivered to cells.

[0123] The systems, methods, compositions, and kits provided herein can, in some embodiments, be employed in concert with the systems, methods, compositions, and kits described in U.S. Patent Application Publication No. US20220402977A1, the content of which is incorporated herein by reference in its entirety. Additionally, the systems, methods, compositions, and kits provided herein can, in some embodiments, be employed in concert with the systems, methods, compositions, and kits described in U.S. Provisional Patent Application No. 63 / 654,437, entitled “CO-DELIVERY OF A TETHERIN ANTAGONIST RESCUES BUDDING OF ENVELOPED NANOPARTICLES (ENPS) IN TETHERIN-EXPRESSING CELLS”, filed May 31, 2024, and in PCT Patent Application No. PCT / US2025 / 022923, entitled “ENGINEERED VIRAL AND MAMMALIAN ESCRT-RECRUITING DOMAINS (ERDS) INDUCE EFFICIENT BUDDING OF ENVELOPED NANOPARTICLES (ENPS) FOR VARIOUS IMMUNOGENS”, filed April 3, 2025, the contents of which are hereby incorporated by reference in their entireties.Adapter Systems

[0124] There are provided, in some embodiments, compositions (e.g., nucleic acid compositions, population(s) of ENPs). In some embodiments, the composition is a vaccine composition. In some embodiments, the composition comprises a nucleic acid composition (e.g., mRNA vaccine, DNA vaccine, a construct).

[0125] In some embodiments, the composition comprises: a nucleic acid composition comprising: (i) a polynucleotide encoding a dimerization fusion protein, wherein the dimerization fusion protein comprises an optional antigenic polypeptide (AP) and a heterologous cytoplasmic tail, and (ii) a polynucleotide encoding an adapter fusion protein comprising an endosomal sorting complex required for transport (ESCRT)-recruiting domain (ERD) and an adapter domain capable of binding the heterologous cytoplasmic tail to form a heterodimer, wherein binding of the adapter domain to the heterologous cytoplasmic tail is capable of recruiting one or more ESCRT proteins to the heterodimer, thereby inducing a plurality of dimerization fusion proteins to self-assemble into an enveloped nanoparticle (ENP) secreted from a cell in which the dimerization fusion protein and adapter fusion protein are expressed, thereby generating a population of ENPs.

[0126] In some embodiments, the composition comprises: a nucleic acid composition comprising (i) n polynucleotides each encoding an nth dimerization fusion protein, wherein n is an integer from 2 to 500 (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80, 90, 100, 110, 120, 128, 130, 140, 150, 160, 170, 180, 190, 200, 210,220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490, 500, or a number or a range between any two of these values), wherein each dimerization fusion protein comprises an optional antigenic polypeptide (AP) and a heterologous cytoplasmic tail, wherein at least two of the dimerization fusion proteins differ with respect to the AP; and (ii) a polynucleotide encoding an adapter fusion protein comprising an endosomal sorting complex required for transport (ESCRT)-recruiting domain (ERD) and an adapter domain capable of binding the heterologous cytoplasmic tail to form a heterodimer, wherein binding of the adapter domain to the heterologous cytoplasmic tail is capable of recruiting one or more ESCRT proteins to the heterodimer, thereby inducing a plurality of dimerization fusion proteins to self-assemble into an enveloped nanoparticle (ENP) secreted from a cell in which the n dimerization fusion proteins and the adapter fusion protein are expressed, thereby generating a population of ENPs.

[0127] In some embodiments, the dimerization fusion proteins are capable of being presented on the surface of a cell in which the dimerization fusion proteins are expressed. In some embodiments, the self-assembly of an ENP: does not require an exogenous nucleic acid other than the nucleic acid composition, and / or does not require any exogenous components other than the dimerization fusion proteins and the adapter fusion proteins.

[0128] In some embodiments, the cell is: a cell of a subject; an in vivo cell, an ex vivo cell, or an in situ cell; and / or an adherent cell or a suspension cell. In some embodiments, upon secretion from a cell of a subject, the ENPs are capable of distributing within one or more tissues of the subject. The one or more tissues can comprise adrenal gland tissue, appendix tissue, bladder tissue, bone, bowel tissue, brain tissue, breast tissue, bronchi, coronal tissue, ear tissue, esophagus tissue, eye tissue, gall bladder tissue, genital tissue, heart tissue, hypothalamus tissue, kidney tissue, large intestine tissue, intestinal tissue, larynx tissue, liver tissue, lung tissue, lymph nodes, mouth tissue, nose tissue, pancreatic tissue, parathyroid gland tissue, pituitary gland tissue, prostate tissue, rectal tissue, salivary gland tissue, skeletal muscle tissue, skin tissue, small intestine tissue, spinal cord, spleen tissue, stomach tissue, thymus gland tissue, trachea tissue, thyroid tissue, ureter tissue, urethra tissue, soft and connective tissue, peritoneal tissue, blood vessel tissue, fat tissue, or any combination thereof. In some embodiments, the ENPs engage a plurality of immune cells in the one or more tissues, thereby mimicking a natural infection.

[0129] Disclosed herein include compositions. In some embodiments, the composition comprises: a population of enveloped nanoparticles (ENPs), wherein each of the ENPs comprises a plurality of the dimerization fusion proteins each comprising a heterologous cytoplasmic tail and optionally, an antigenic polypeptide (AP). The ENPs can be derived from expression of any of the nucleic acid compositions of the disclosure.

[0130] The adapter fusion protein can comprise, from N-terminus to C-terminus: the adapter domain, an optional linker, and the ERD. In some embodiments, the linker: is a flexible linker, a rigid linker, or a hybrid linker; is hydrophilic or hydrophobic; is between 1 and 250 amino acids (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26,27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52,53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 150, 200, 250, or a number between any two of these values, amino acids in length); or comprises one or more flexible amino acid residues. The linker can comprise about 1 to about 250 flexible amino acid residues. In some embodiments, the flexible amino acid residues comprise glycine, serine, or a combination thereof. In some embodiments, the linker comprises 3 repeating amino acid subunits or more.

[0131] In some embodiments, the heterologous cytoplasmic tail is derived from or comprises a cytoplasmic tail of a cell surface protein. In some embodiments, the adapter domain is (1) capable of binding the heterologous cytoplasmic tail derived from or comprising the cytoplasmic tail of said cell surface protein and (2) capable of targeting the adapter fusion protein to the plasma membrane. In some embodiments, the cell surface protein is or is derived from a human protein, a non-human mammalian protein, an avian protein, or a reptile protein and / or the adapter domain is or is derived from a human protein, a non-human mammalian protein, an avian protein, or a reptile protein.

[0132] In some embodiments, the heterologous cytoplasmic tail comprises or is derived from a cell surface protein of an immune cell. In some embodiments, the adapter domain is capable of binding to a portion of the cell surface protein of said immune cell, e.g., the cytoplasmic tail or other cytosolic portion of the cell surface protein. In some embodiments, the heterologous cytoplasmic tail comprises or is derived from a cytoplasmic tail (CT) of CD4 or CD8; and / or the adapter domain comprises or is derived from at least a portion of Lek tyrosine kinase. The CD4 CT can comprise the sequence of SEQ ID NO: 95 or SEQ ID NO: 96. The CD4 CT can comprise a sequence having one, two, or three mismatches relative to the sequence of SEQ ID NO: 95 or SEQ ID NO: 96. The CD8 CT can comprise the sequence of SEQ ID NO: 101 or a sequence having one, two, or three mismatches relative to the sequence of SEQ ID NO: 101.

[0133] The adapter domain can comprise the sequence of SEQ ID NO: 43 or SEQ ID NO: 97. The adapter domain can comprise a sequence having one, two, or three mismatches relative to the sequence of SEQ ID NO: 43 or SEQ ID NO: 97. The heterologous cytoplasmic tail of CD4 can comprise the sequence of SEQ ID NO: 98, SEQ ID NO: 99, or SEQ ID NO: 100. The heterologous cytoplasmic tail of CD4 can comprise a sequence having one, two, or threemismatches relative to the sequence of SEQ ID NO: 98, SEQ ID NO: 99, or SEQ ID NO: 100.

[0134] The heterologous cytoplasmic tail and / or the adapter domain can be modified (e.g., by mutation) to form disulfide bonds between the heterologous cytoplasmic tail and the adapter domain. In some embodiments: the heterologous cytoplasmic tail of CD4 comprises a sequence having one, two, or three mismatches relative to the sequence of SEQ ID NO: 102, and the adapter domain comprises a sequence having one, two, or three mismatches relative to the sequence of SEQ ID NO: 105; the heterologous cytoplasmic tail of CD4 comprises a sequence having one, two, or three mismatches relative to the sequence of SEQ ID NO: 103, and the adapter domain comprises a sequence having one, two, or three mismatches relative to the sequence of SEQ ID NO: 106; or the heterologous cytoplasmic tail of CD4 comprises a sequence having one, two, or three mismatches relative to the sequence of SEQ ID NO: 104, and the adapter domain comprises a sequence having one, two, or three mismatches relative to the sequence of SEQ ID NO: 107. In some embodiments: the heterologous cytoplasmic tail of CD4 comprises the sequence of SEQ ID NO: 102, and the adapter domain comprises the sequence of SEQ ID NO: 105; the heterologous cytoplasmic tail of CD4 comprises the sequence of SEQ ID NO: 103, and the adapter domain comprises the sequence of SEQ ID NO: 106; or the heterologous cytoplasmic tail of CD4 comprises the sequence of SEQ ID NO: 104, and the adapter domain comprises the sequence of SEQ ID NO: 107.

[0135] In some embodiments, the composition further comprises: a polynucleotide encoding a second adapter fusion protein comprising an endosomal sorting complex required for transport (ESCRT)-recruiting domain (ERD) and an adapter domain capable of binding the heterologous cytoplasmic tail to form a heterodimer, wherein the adapter domain of the second adapter fusion protein is the same or different from the adapter domain of the adapter fusion protein. In some embodiments, the ERD of the second adapter fusion protein is the same or different from the ERD of the adapter fusion protein. In some embodiments, the adapter domain and the second adapter domain are both capable of binding the heterologous cytoplasmic tail. The adapter domain of the second adapter fusion protein can comprise the sequence of SEQ ID NO: 109 (or a sequence having one, two, or three mismatches relative to the sequence of SEQ ID NO: 109) and the adapter domain of the adapter fusion protein can comprise the sequence of SEQ ID NO: 43 or SEQ ID NO: 97 (or a sequence having one, two, or three mismatches relative to the sequence of SEQ ID NO: 43 or SEQ ID NO: 97).

[0136] In some embodiments, the heterologous cytoplasmic tail and / or the adapter domain comprise or are derived from a non-mammalian protein. In some embodiments, the heterologous cytoplasmic tail comprises or is derived from G. gallus CD4 CT. The heterologous cytoplasmic tail can comprise the sequence of SEQ ID NO: 110 (or a sequence having one, two,or three mismatches relative to the sequence of SEQ ID NO: 110). In some embodiments, the adapter domain comprises or is derived from at least a portion of G. gallus Lek tyrosine kinase. The adapter domain can comprise the sequence of SEQ ID NO: 113 (or a sequence having one, two, or three mismatches relative to the sequence of SEQ ID NO: 113).

[0137] In some embodiments, the heterologous cytoplasmic tail comprises or is derived from P. vitticeps CD4 CT. The heterologous cytoplasmic tail can comprise the sequence of SEQ ID NO: 111 (or a sequence having one, two, or three mismatches relative to the sequence of SEQ ID NO: 111). In some embodiments, the adapter domain comprises or is derived from at least a portion of P. vitticeps Lek tyrosine kinase. The adapter domain can comprise the sequence of SEQ ID NO: 114 (or a sequence having one, two, or three mismatches relative to the sequence of SEQ ID NO: 114). In some embodiments, the heterologous cytoplasmic tail comprises or is derived from N scutatus CD4 CT. The heterologous cytoplasmic tail can comprise the sequence of SEQ ID NO: 112 (or a sequence having one, two, or three mismatches relative to the sequence of SEQ ID NO: 112). In some embodiments, the adapter domain comprises or is derived from at least a portion of N scutatus Lek tyrosine kinase. The adapter domain can comprise the sequence of SEQ ID NO: 115 (or a sequence having one, two, or three mismatches relative to the sequence of SEQ ID NO: 115).

[0138] The dimerization fusion protein can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to an amino acid sequence selected from the group consisting of SEQ ID NOs: 41-42, 45, 48-59, 63, 68-69, and 83-85. The adapter fusion protein can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to an amino acid sequence selected from the group consisting of SEQ ID NOs: 44, 46, 60-62, 64-65, 70-82, and 86-88.

[0139] The dimerization fusion protein can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to the sequence of SEQ ID NO: 51 ; and the adapter fusion protein can comprise an amino acid sequence having at least 65%, 70%,75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to an amino acid sequence selected from the group consisting of SEQ ID NOs: 44, 64, 77, and 80-82.

[0140] The dimerization fusion protein can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to the sequence of SEQ ID NO: 51 ; and the adapter fusion protein can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to the sequence of SEQ ID NO: 44.

[0141] The dimerization fusion protein can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to the sequence of SEQ ID NO: 51 ; and the adapter fusion protein can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to the sequence of SEQ ID NO: 64.

[0142] The dimerization fusion protein can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to the sequence of SEQ ID NO: 51 ; and the adapter fusion protein can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to the sequence of SEQ ID NO: 77.

[0143] The dimerization fusion protein can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to the sequence of SEQ ID NO: 51 ; and the adapter fusion protein can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to the sequence of SEQ ID NO: 80.

[0144] The dimerization fusion protein can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to the sequence of SEQ ID NO: 51 ; and the adapter fusion protein can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to the sequence of SEQ ID NO: 81.

[0145] The dimerization fusion protein can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to the sequence of SEQ ID NO: 51 ; and the adapter fusion protein can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to the sequence of SEQ ID NO: 82.

[0146] The dimerization fusion protein can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to the sequence of SEQ ID NO:69; and the adapter fusion protein can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to an amino acid sequence selected from the group consisting of SEQ ID NOs: 44, 64, 77, and 80-82.

[0147] The dimerization fusion protein can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to the sequence of SEQ ID NO: 69; and the adapter fusion protein can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to the sequence of SEQ ID NO: 44.

[0148] The dimerization fusion protein can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to the sequence of SEQ ID NO: 69; and the adapter fusion protein can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to the sequence of SEQ ID NO: 64.

[0149] The dimerization fusion protein can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to the sequence of SEQ ID NO: 69; and the adapter fusion protein can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or anumber or a range between any two of these values) to the sequence of SEQ ID NO: 77.

[0150] The dimerization fusion protein can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to the sequence of SEQ ID NO: 69; and the adapter fusion protein can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to the sequence of SEQ ID NO: 80.

[0151] The dimerization fusion protein can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to the sequence of SEQ ID NO: 69; and the adapter fusion protein can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to the sequence of SEQ ID NO: 81.

[0152] The dimerization fusion protein can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to the sequence of SEQ ID NO: 69; and the adapter fusion protein can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to the sequence of SEQ ID NO: 82.

[0153] The dimerization fusion protein can comprise, from N-terminus to C-terminus: the antigenic polypeptide, an optional linker, and the heterologous cytoplasmic tail. In some embodiments, the endogenous cytoplasmic tail of an antigen (e.g., Spike protein), is replaced with the heterologous cytoplasmic tail. In some embodiments, the heterologous cytoplasmic tail isfused to the endogenous cytoplasmic tail of the antigenic protein, optionally, separated by a linker. The dimerization fusion protein can comprise an endogenous cytoplasmic tail N-terminal to the heterologous cytoplasmic tail. In some embodiments, the dimerization fusion protein comprises, from C-terminus toN-terminus: the antigenic polypeptide, an optional linker, and the heterologous cytoplasmic tail. In some embodiments, the dimerization fusion protein comprises an endogenous cytoplasmic tail C-terminal to the heterologous cytoplasmic tail.

[0154] The adapter fusion protein can comprise, from N-terminus to C-terminus: a heterologous membrane-targeting domain, an optional first linker, the adapter domain, an optional second linker, and the ERD. In some embodiments, the linker, the first linker, and / or the second linker: is a flexible linker, a rigid linker, or a hybrid linker; is hydrophilic or hydrophobic; is between 1 and 250 amino acids (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19,20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45,46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71,72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,98, 99, 100, 150, 200, 250, or a number between any two of these values, amino acids in length); comprises one or more flexible amino acid residues, optionally about 1 to about 250 flexible amino acid residues, further optionally the flexible amino acid residues comprise glycine, serine, or a combination thereof. In some embodiments, the linker comprises 3 repeating amino acid subunits or more.

[0155] In some embodiments, an adapter domain comprises endogenous sequence capable of targeting the adapter fusion protein to the plasma membrane. In some embodiments, the adapter fusion protein comprises heterologous sequence for targeting the adapter fusion protein to the plasma membrane (e.g., a heterologous membrane targeting domain). In some embodiments, the heterologous membrane targeting domain comprises or is derived from at least a portion of LAT, PAG, LCK, FYN, LAX, CD2, CD3, CD4, CD5, CD7, CD8a, PD1, SRC, or LYN. In some embodiments, the heterologous membrane targeting domain comprises or is derived from FYN. The heterologous membrane targeting domain can comprise the sequence of SEQ ID NO: 118 or a sequence having one, two, or three mismatches relative to the sequence of SEQ ID NO: 118.

[0156] In some embodiments, the heterologous cytoplasmictail and / or the adapter domain are each selected from the group comprising DHD9 heterodimer a, DHD13 XAAA heterodimer a, DHD13 XAXA heterodimer a, DHD13 XAAX heterodimer a, DHD13_2:341 heterodimer a, DHD13 AAAA heterodimer a, DHD13 BAAA heterodimer a, DHD13_4: 123 heterodimer a, DHD13_1 :234 heterodimer a, DHD15 heterodimer a, DHD20 heterodimer a, DHD21 heterodimer a, DHD25 heterodimer a, DHD27 heterodimer a, DHD30 heterodimer a,DHD33 heterodimer a, DHD34 XAAXA heterodimer a, DHD34 XAXXA heterodimer a, DHD34 XAAAA heterodimer a, DHD36 heterodimer a, DHD37 ABXB heterodimer a, DHD37 BBBB heterodimer a, DHD37 XBXB heterodimer a, DHD37 AXXB heterodimer a, DHD37_3: 124 heterodimer a, DHD37_1 :234 heterodimer a, DHD37 AXBB heterodimer a, DHD37 XBBA heterodimer a, DHD39 heterodimer a, DHD40 heterodimer a, DHD43 heterodimer a, DHD65 heterodimer a, DHD70 heterodimer a, DHD88 heterodimer a, DHD89 heterodimer a, DHD90 heterodimer a, DHD91 heterodimer a, DHD92 heterodimer a, DHD93 heterodimer a, DHD94 heterodimer a, DHD94_3:214 heterodimer a, DHD94_2: 143 heterodimer a, DHD95 heterodimer a, DHD96 heterodimer a, DHD97 heterodimer a, DHD98 heterodimer a, DHD99 heterodimer a, DHD100 heterodimer a, DHD101 heterodimer a, DHD102 heterodimer a, DHD102_l :243 heterodimer a, DHD103 heterodimer a, DHD103_l :423 heterodimer a, DHD104 heterodimer a, DHD105 heterodimer a, DHD106 heterodimer a, DHD107 heterodimer a, DHD108 heterodimer a, DHD109 heterodimer a, DHD110 heterodimer a, DHD111 heterodimer a, DHD112 heterodimer a, DHD113 heterodimer a, DHD114 heterodimer a, DHD115 heterodimer a, DHD116 heterodimer a, DHD117 heterodimer a, DHD118 heterodimer a, DHD119 heterodimer a, DHD120 heterodimer a, DHD121 heterodimer a, DHD122 heterodimer a, DHD123 heterodimer a, DHD124 heterodimer a, DHD125 heterodimer a, DHD126 heterodimer a, DHD127 heterodimer a, DHD128 heterodimer a, DHD129 heterodimer a, DHD130 heterodimer a, DHD145 heterodimer a, DHD146 heterodimer a, DHD147 heterodimer a, DHD1 heterodimer a, DHD2 heterodimer a, DHD3 heterodimer a, DHD4 heterodimer a, DHD5 heterodimer a, DHD6 heterodimer a, DHD7 heterodimer a, DHD8 heterodimer a, DHD16 heterodimer a, DHD18 heterodimer a, DHD19 heterodimer a, DHD22 heterodimer a, DHD23 heterodimer a, DHD24 heterodimer a, DHD26 heterodimer a, DHD28 heterodimer a, DHD29 heterodimer a, DHD31 heterodimer a, DHD32 heterodimer a, DHD38 heterodimer a, DHD60 heterodimer a, DHD63 heterodimer a, DHD66 heterodimer a, DHD67 heterodimer a, DHD69 heterodimer a, DHD71 heterodimer a, DHD72 heterodimer a, DHD73 heterodimer a, DHD148 heterodimer a, DHD149 heterodimer a, DHD150 heterodimer a, DHD151 heterodimer a, DHD152 heterodimer a, DHD153 heterodimer a, DHD154 heterodimer a, DHD155 heterodimer a, DHD156 heterodimer a, DHD157 heterodimer a, DHD158 heterodimer a, DHD159 heterodimer a, DHD160 heterodimer a, DHD161 heterodimer a, DHD162 heterodimer a, DHD163 heterodimer a, DHD164 heterodimer a, DHD165 heterodimer a, DHD166 heterodimer a, DHS17 heterodimer a, DHD17 heterodimer a, DHD131 heterodimer a, DHD132 heterodimer a, DHD133 heterodimer a, DHD134 heterodimer a, DHD135 heterodimer a, DHD136 heterodimer a, DHD137 heterodimer a, DHD138 heterodimer a, DHD139 heterodimer a, DHD140 heterodimer a, DHD141 heterodimer a, DHD142 heterodimer a, DHD143 heterodimera, DHD144 heterodimer a, DHD9 heterodimer b, DHD13 XAAA heterodimer b, DHD13 XAXA heterodimer b, DHD13 XAAX heterodimer b, DHD13_2:341 heterodimer b, DHD13 AAAA heterodimer b, DHD13 BAAA heterodimer b, DHD13_4: 123 heterodimer b, DHD13_1 :234 heterodimer b, DHD15 heterodimer b, DHD20 heterodimer b, DHD21 heterodimer b, DHD25 heterodimer b, DHD27 heterodimer b, DHD30 heterodimer b, DHD33 heterodimer b, DHD34 XAAXA heterodimer b, DHD34 XAXXA heterodimer b, DHD34 XAAAA heterodimer b, DHD36 heterodimer b, DHD37 ABXB heterodimer b, DHD37 BBBB heterodimer b, DHD37_XBXB heterodimer b, DHD37 AXXB heterodimer b, DHD37_3: 124 heterodimer b, DHD37_1 :234 heterodimer b, DHD37 AXBB heterodimer b, DHD37 XBBA heterodimer b, DHD39 heterodimer b, DHD40 heterodimer b, DHD43 heterodimer b, DHD65 heterodimer b, DHD70 heterodimer b, DHD88 heterodimer b, DHD89 heterodimer b, DHD90 heterodimer b, DHD91 heterodimer b, DHD92 heterodimer b, DHD93 heterodimer b, DHD94 heterodimer b, DHD94_3:214 heterodimer b, DHD94_2: 143 heterodimer b, DHD95 heterodimer b, DHD96 heterodimer b, DHD97 heterodimer b, DHD98 heterodimer b, DHD99 heterodimer b,DHD100 heterodimer b, DHD101 heterodimer b, DHD102 heterodimer b, DHD102_l :243 heterodimer b, DHD103 heterodimer b, DHD103_l :423 heterodimer b, DHD104 heterodimer b, DHD105 heterodimer b, DHD106 heterodimer b, DHD107 heterodimer b, DHD108 heterodimer b, DHD109 heterodimer b, DHD110 heterodimer b, DHD111 heterodimer b, DHD112 heterodimer b, DHD113 heterodimer b, DHD114 heterodimer b, DHD115 heterodimer b, DHD1 16 heterodimer b, DHD117 heterodimer b, DHD118 heterodimer b, DHD119 heterodimer b, DHD120 heterodimer b, DHD121 heterodimer b, DHD122 heterodimer b, DHD123 heterodimer b, DHD124 heterodimer b, DHD125 heterodimer b, DHD126 heterodimer b, DHD127 heterodimer b, DHD128 heterodimer b, DHD129 heterodimer b, DHD130 heterodimer b, DHD145 heterodimer b, DHD146 heterodimer b, DHD147 heterodimer b, DHD1 heterodimer b, DHD2 heterodimer b, DHD3 heterodimer b, DHD4 heterodimer b, DHD5 heterodimer b, DHD6 heterodimer b, DHD7 heterodimer b, DHD8 heterodimer b, DHD16 heterodimer b, DHD18 heterodimer b, DHD19 heterodimer b, DHD22 heterodimer b, DHD23 heterodimer b,DHD24 heterodimer b, DHD26 heterodimer b, DHD28 heterodimer b, DHD29 heterodimer b,DHD31 heterodimer b, DHD32 heterodimer b, DHD38 heterodimer b, DHD60 heterodimer b,DHD63 heterodimer b, DHD66 heterodimer b, DHD67 heterodimer b, DHD69 heterodimer b,DHD71 heterodimer b, DHD72 heterodimer b, DHD73 heterodimer b, DHD148 heterodimer b, DHD149 heterodimer b, DHD150 heterodimer b, DHD151 heterodimer b, DHD152 heterodimer b, DHD153 heterodimer b, DHD154 heterodimer b, DHD155 heterodimer b, DHD156 heterodimer b, DHD157 heterodimer b, DHD158 heterodimer b, DHD159 heterodimer b, DHD160 heterodimer b, DHD161 heterodimer b, DHD162 heterodimer b, DHD163 heterodimerb, DHD164 heterodimer b, DHD165 heterodimer b, DHD166 heterodimer b, DHS17 heterodimer b, DHD17 heterodimer b, DHD131 heterodimer b, DHD132 heterodimer b, DHD133 heterodimer b, DHD134 heterodimer b, DHD135 heterodimer b, DHD136 heterodimer b, DHD137 heterodimer b, DHD138 heterodimer b, DHD139 heterodimer b, DHD140 heterodimer b, DHD141 heterodimer b, DHD142 heterodimer b, DHD143 heterodimer b, DHD144 heterodimer b, portions thereof, derivatives thereof, or any combination thereof.

[0157] In some embodiments, the heterologous cytoplasmic tail and / or the adapter domain comprises or is derived from SYNZIP1, SYNZIP2, SYNZIP3, SYNZIP4, SYNZIP5, SYNZIP6, SYNZIP7, SYNZIP8, SYNZIP9, SYNZIP10, SYNZIP11, SYNZIP12, SYNZIP13, SYNZIP14, SYNZIP15, SYNZIP16, SYNZIP17, SYNZIP18, SYNZIP19, SYNZIP20, SYNZIP21, SYNZIP22, SYNZIP23, BATF, FOS, ATF4, BACH1, JUNE), NFE2L3, AZip, BZip, a PDZ domain ligand, an SH3 domain, a PDZ domain, a GTPase binding domain, a leucine zipper domain, an SH2 domain, a PTB domain, an FHA domain, a WW domain, a 14-3- 3 domain, a death domain, a caspase recruitment domain, a bromodomain, a chromatin organization modifier, a shadow chromo domain, an F-box domain, a HECT domain, a RING finger domain, a sterile alpha motif domain, a glycine-tyrosine-phenylalanine domain, a SNAP domain, a VHS domain, an ANK repeat, an armadillo repeat, a WD40 repeat, an MH2 domain, a calponin homology domain, a Dbl homology domain, a gelsolin homology domain, a PB 1 domain, a SOCS box, an RGS domain, a Toll / IL-1 receptor domain, a tetratricopeptide repeat, a TRAF domain, a Bcl-2 homology domain, a coiled-coil domain, a bZIP domain, portions thereof, variants thereof, or any combination thereof.

[0158] In some embodiments, the heterologous cytoplasmic tail comprises or is derived from ACIDpl or BASEp 1 or the heterologous cytoplasmic tail comprises or is derived from N5 or N6. The heterologous cytoplasmic tail can comprise the sequence of any one of SEQ ID NOs: 116-117 and 121-122. The heterologous cytoplasmic tail can comprise a sequence having one, two, or three mismatches relative to the sequence of any one of SEQ ID NOs: 116-117 and 121-122. In some embodiments, the adapter domain comprises or is derived from ACIDpl or BASEp 1 or the heterologous cytoplasmic tail comprises or is derived from N5 or N6. The adapter domain can comprise the sequence of any one of SEQ ID NOs: 116-117 and 121-122. The adapter domain can comprise a sequence having one, two, or three mismatches relative to any one of SEQ ID NOs: 116-117 and 121-122.

[0159] The dimerization fusion protein can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%,100%, or a number or a range between any two of these values) to an amino acid sequence selected from the group consisting of SEQ ID NOs: 89-90 and 93. The adapter fusion protein can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to an amino acid sequence selected from the group consisting of SEQ ID NOs: 91-92 and 94.

[0160] The dimerization fusion protein can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to the sequence of SEQ ID NO: 93; and the adapter fusion protein can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to the sequence of SEQ ID NO: 94.

[0161] In some embodiments, (i) the polynucleotide encoding the dimerization fusion protein or the n polynucleotides each encoding an nth dimerization fusion protein, and (ii) the polynucleotide encoding the adapter fusion protein, are each present in a different nucleic acid molecule. The amount of (i) the polynucleotide encoding the dimerization fusion protein or the n polynucleotides each encoding an nth dimerization fusion protein; and (ii) the polynucleotide encoding the adapter fusion protein present in the composition can vary. In some embodiments, the amount of (i) the polynucleotide encoding the dimerization fusion protein or the n polynucleotides each encoding an nth dimerization fusion protein; and (ii) the polynucleotide encoding the adapter fusion protein, are present in the composition at a molar ratio of about 9:1, 5:1, or 1:1 (e.g., 1:1, 1.1:1, 1.2:1, 1.3:1, 1.4:1, 1.5:1, 1.6:1, 1.7:1, 1.8:1, 1.9:1, 2:1, 2.5:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, 20:1, 21:1, 22:1, 23:1, 24:1, 25:1, 26:1, 27:1, 28:1, 29:1, 30:1, 31:1, 32:1, 33:1, 34:1, 35:1, 36:1, 37:1, 38:1, 39:1,40:1, 41:1, 42:1, 43:1, 44:1, 45:1, 46:1, 47:1, 48:1, 49:1, 50:1, 51:1, 52:1, 53:1, 54:1, 55:1, 56:1,57:1, 58:1, 59:1, 60:1, 61:1, 62:1, 63:1, 64:1, 65:1, 66:1, 67:1, 68:1, 69:1, 70:1, 71:1, 72:1, 73:1,74:1, 75:1, 76:1, 77:1, 78:1, 79:1, 80:1, 81:1, 82:1, 83:1, 84:1, 85:1, 86:1, 87:1, 88:1, 89:1, 90:1,91:1, 92:1, 93:1, 94:1, 95:1, 96:1, 97:1, 98:1, 99:1, 100:1, or a number or a range between any of these values). In some embodiments, (i) the polynucleotide encoding the dimerization fusionprotein or the n polynucleotides each encoding an nth dimerization fusion protein, and (ii) the polynucleotide encoding the adapter fusion protein, are present in the same nucleic acid molecule.

[0162] In some embodiments, the cell expressing the dimerization fusion protein or the plurality of dimerization fusion proteins and the adapter fusion protein exhibits at least a 2- fold (e.g., 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 20-fold, 30-fold, 40- fold, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold, 100-fold, or a number or a range between any of these values) increase in AP expression at the surface of the cell, relative to a cell expressing a fusion protein or a plurality of fusion proteins comprising an AP and an ERD and does not express the adapter fusion protein. In some embodiments, the production of ENPs from the cell expressing the dimerization fusion protein or the plurality of dimerization fusion proteins and the adapter fusion protein is increased by at least 2-fold (e.g., 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8- fold, 9-fold, 10-fold, 20-fold, 30-fold, 40-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold, 100- fold, or a number or a range between any of these values), relative to a cell expressing a fusion protein or a plurality of fusion proteins comprising an AP and an ERD and does not express the adapter fusion protein.

[0163] In some embodiments, the nucleic acid composition further comprises a polynucleotide comprising or encoding a tetherin inhibitor. In some embodiments, the tetherin inhibitor is capable of modulating expression, concentration, localization, stability, and / or activity of tetherin. In some embodiments, the tetherin inhibitor comprises a dsRNA, an siRNA, an shRNA, a pre-miRNA, a pri-miRNA, a miRNA, an stRNA, an IncRNA, a piRNA, a snoRNA, or a protein. In some embodiments, one or more of (i) the polynucleotide comprising or encoding the tetherin inhibitor, (ii) the polynucleotide encoding the dimerization fusion protein or the n polynucleotides each encoding an nth dimerization fusion protein, and (iii) the polynucleotide encoding the adapter fusion protein, are present in a same or a different nucleic acid molecule. In some embodiments, the amount of (i) the polynucleotide comprising or encoding the tetherin inhibitor; and (ii) the polynucleotide encoding the dimerization fusion protein or the n polynucleotides each encoding an nth dimerization fusion protein, and / or the polynucleotide encoding the adapter fusion protein, are present in the composition at a molar ratio of about 1:1, 1:5 or 1:25. In some embodiments, the amount of (i) the polynucleotide comprising or encoding the tetherin inhibitor; and (ii) the polynucleotide encoding the dimerization fusion protein or the n polynucleotides each encoding an nth dimerization fusion protein, and / or the polynucleotide encoding the adapter fusion protein, are present in the composition at a molar ratio of about 1:1, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9, 1:2, 1:2.5, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, 1:20, 1:21, 1:22, 1:23, 1:24, 1:25, 1:26, 1:27, 1:28, 1:29, 1:30, 1:31, 1:32, 1:33, 1:34, 1:35, 1:36, 1:37, 1:38, 1:39, 1:40, 1:41, 1:42, 1:43,1 :44, 1 :45, 1 :46, 1 :47, 1 :48, 1 :49, 1 :50, 1 :51, 1 :52, 1 :53, 1 :54, 1 :55, 1 :56, 1 :57, 1 :58, 1 :59, 1 :60,1 :61, 1 :62, 1 :63, 1 :64, 1 :65, 1 :66, 1 :67, 1 :68, 1 :69, 1 :70, 1 :71, 1 :72, 1 :73, 1 :74, 1 :75, 1 :76, 1 :77,1 :78, 1 :79, 1 :80, 1 :81, 1 :82, 1 :83, 1 :84, 1 :85, 1 :86, 1 :87, 1 :88, 1 :89, 1 :90, 1 :91, 1 :92, 1 :93, 1 :94,1 :95, 1 :96, 1 :97, 1 :98, 1 :99, or 1 : 100 or a number or a range between any of these values. In some embodiments, the polynucleotide comprising or encoding the tetherin inhibitor and the polynucleotide encoding the dimerization fusion protein are present in the same nucleic acid. In some embodiments, the polynucleotide comprising or encoding the tetherin inhibitor and the polynucleotide encoding the adapter fusion protein are present in the same nucleic acid. For example, the polynucleotide encoding the tetherin inhibitor and the polynucleotide encoding the dimerization fusion protein or the adapter fusion protein can be operably linked to a tandem gene expression element (e.g., an internal ribosomal entry site (IRES), foot-and-mouth disease virus 2A peptide (F2A), equine rhinitis A virus 2A peptide (E2A), porcine teschovirus 2A peptide (P2A) or Thosea asigna virus 2A peptide (T2A), or any combination thereof).

[0164] In some embodiments, the tetherin inhibitor comprises or is derived from a viral protein, optionally the virus is HIV-1, HIV-2, SIV, Ebola virus, KSHV, SARS CoV, or SARS-CoV-2. In some embodiments, the tetherin inhibitor comprises HIV-1 Vpu protein, KSHV K5 protein, SARS-CoV-2 ORF7a, HIV-2 Env, Ebola GP, SIV Env, SIV Vpu, SIV Nef, or any portions, variants or derivatives thereof. In some embodiments, the tetherin inhibitor comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to an amino acid sequence selected from the group consisting of SEQ ID NOs: 132-134. In some embodiments, the tetherin inhibitor comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to the sequence of SEQ ID NO: 132. In some embodiments, presence or expression of the tetherin inhibitor in the cell results in an increase in ENP production by the cell by at least 2-fold (e.g., 2-fold, 3-fold, 4-fold, 5-fold, 6- fold, 7-fold, 8-fold, 9-fold, 10-fold, 20-fold, 30-fold, 40-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold, 100-fold, or a number or a range between any of these values), relative to a cell that does not comprise or express the tetherin inhibitor.Co-localization Systems

[0165] There are provided, in some embodiments, compositions (e.g., nucleic acidcompositions, population(s) of ENPs). In some embodiments, the composition is a vaccine composition. In some embodiments, the composition comprises a nucleic acid composition (e.g., mRNA vaccine, DNA vaccine, a construct).

[0166] Disclosed herein include compositions. In some embodiments, the composition comprises: a nucleic acid composition comprising: (i) a polynucleotide encoding a recombinant protein comprising an optional antigenic polypeptide (AP) and a transmembrane domain, and (ii) a polynucleotide encoding a chimeric protein comprising (e.g., from N terminus to C terminus): a) a signal peptide, b) a cell surface domain, c) a transmembrane domain, and d) a cytoplasmic domain; wherein the recombinant protein and / or the chimeric protein further comprises an endosomal sorting complex required for transport (ESCRT)-recruiting domain (ERD), optionally the recombinant protein further comprises a cytoplasmic domain; and wherein the recombinant protein and the chimeric protein are capable of co-localizing to a site on the plasma membrane of the cell, and wherein the ERD is capable of recruiting one or more ESCRT proteins to the site on the plasma membrane, thereby inducing a plurality of recombinant proteins and chimeric proteins to self-assemble into an enveloped nanoparticle (ENP) secreted from a cell in which the recombinant protein and chimeric protein are expressed, thereby generating a population of ENPs, optionally the cytoplasmic domain of the recombinant protein and the cytoplasmic domain of the chimeric protein are capable of co-localizing the recombinant protein and the chimeric protein to a site on the plasma membrane of the cell, optionally the transmembrane domain of the recombinant protein and the transmembrane domain of the chimeric protein are capable of colocalizing the recombinant protein and the chimeric protein to a site on the plasma membrane of the cell.

[0167] Disclosed herein include compositions. In some embodiments, the composition comprises: a nucleic acid composition comprising: (i) n polynucleotides each encoding a recombinant protein comprising an optional antigenic polypeptide (AP) and a transmembrane domain, wherein at least two of the recombinant proteins differ with respect to the AP, wherein n is an integer from 2 to 500 (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80, 90, 100, 110, 120, 128, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490, 500, or a number or a range between any two of these values); and (ii) z polynucleotides each encoding a chimeric protein comprising (e.g., from N terminus to C terminus): a) a signal peptide, b) a cell surface domain, wherein at least two of the chimeric proteins differ with respect to the cell surface domain, c) a transmembrane domain, and d) a cytoplasmic domain wherein z is an integer from 1 to 500 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80, 90, 100, 110, 120, 128, 130,140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490, 500, or a number or a range between any two of these values), wherein the nth recombinant proteins and / or the zth chimeric proteins further comprise an endosomal sorting complex required for transport (ESCRT)-recruiting domain (ERD), optionally the nth recombinant proteins further comprise a cytoplasmic domain, and wherein each of the nth recombinant protein and each of the zth chimeric protein are capable of co-localizing to a site on the plasma membrane of the cell, and the ERD is capable of recruiting one or more ESCRT proteins to the site on the plasma membrane, thereby inducing a plurality of recombinant proteins and chimeric proteins to self-assemble into an enveloped nanoparticle (ENP) secreted from the cell in which the recombinant proteins and chimeric proteins are expressed, thereby generating a population of ENPs, optionally the cytoplasmic domain of each of the nth recombinant protein and the cytoplasmic domain of each of the zth chimeric protein are capable of co-localizing each of the nth recombinant protein and each of the zth chimeric protein to a site on the plasma membrane of the cell, optionally the transmembrane domain of each of the nth recombinant protein and the transmembrane domain of each of the zth chimeric protein are capable of co-localizing each of the nth recombinant protein and each of the zth chimeric protein to a site on the plasma membrane of the cell.

[0168] In some embodiments, the plurality of recombinant proteins and the plurality of chimeric proteins are capable of being presented on the surface of the cell in which the plurality of recombinant proteins and the plurality of chimeric proteins are expressed. In some embodiments, the self-assembly of an ENP: does not require an exogenous nucleic acid other than the nucleic acid composition, and / or does not require any exogenous components other than the recombinant protein and the chimeric protein.

[0169] In some embodiments, the cell is: a cell of a subject; an in vivo cell, an ex vivo cell, or an in situ cell; and / or an adherent cell or a suspension cell. In some embodiments, upon secretion from a cell of a subject, the ENPs are capable of distributing within one or more tissues of the subject. The one or more tissues can comprise adrenal gland tissue, appendix tissue, bladder tissue, bone, bowel tissue, brain tissue, breast tissue, bronchi, coronal tissue, ear tissue, esophagus tissue, eye tissue, gall bladder tissue, genital tissue, heart tissue, hypothalamus tissue, kidney tissue, large intestine tissue, intestinal tissue, larynx tissue, liver tissue, lung tissue, lymph nodes, mouth tissue, nose tissue, pancreatic tissue, parathyroid gland tissue, pituitary gland tissue, prostate tissue, rectal tissue, salivary gland tissue, skeletal muscle tissue, skin tissue, small intestine tissue, spinal cord, spleen tissue, stomach tissue, thymus gland tissue, trachea tissue, thyroid tissue, ureter tissue, urethra tissue, soft and connective tissue, peritoneal tissue, blood vessel tissue, fat tissue, or any combination thereof. In some embodiments, the ENPs engage aplurality of immune cells in the one or more tissues, thereby mimicking a natural infection.

[0170] Disclosed herein include compositions. In some embodiments, the composition comprises: a population of enveloped nanoparticles (ENPs), wherein each of the ENPs comprises a plurality of recombinant proteins comprising a transmembrane domain and an optional antigenic polypeptide (AP), and a plurality of chimeric proteins comprising: a) a signal peptide, b) a cell surface domain, c) a transmembrane domain, and d) a cytoplasmic domain, wherein the plurality of recombinant proteins and / or the plurality of chimeric proteins further comprise an endosomal sorting complex required fortransport (ESCRT)-recruiting domain (ERD), optionally the plurality of recombinant proteins further comprise a cytoplasmic domain. The ENPs can be derived from expression of any of the nucleic acid compositions of the disclosure.

[0171] The chimeric protein can comprise a signal peptide. The signal peptide can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to an amino acid of SEQ ID NO: 127 or a sequence having one, two, or three mismatches relative to the sequence of SEQ ID NO: 127.

[0172] The chimeric protein can comprise a cell surface domain (e.g., an extracellular domain). The cell surface domain can comprise an immune-cell targeting polypeptide or a second antigenic polypeptide sequence. The second antigenic polypeptide sequence can comprise any of the antigenic sequences disclosed herein. In some embodiments, the immune-cell targeting polypeptide comprises or is derived from one or more of complement component 3d (C3d), flagellin, an influenza HA molecule, a parainfluenza FiN molecule, a Venezuelan equine encephalitis (VEE) glycoprotein molecule, a mannose receptor molecule, a mammalian toll- like receptor (TLR) ligand molecule, a MIP-1 alpha molecule, a RANTES MIP-1 beta molecule, a GM-CSF molecule, a Flt3 ligand molecule, a CD40 ligand molecule, a Prevotella intermedia glycoprotein, a respiratory syncytial virus protein F, a fibronectin A domain, fibrinogen, a measles virus HA protein, and Pam2Cys lipoprotein / lipopeptide (MALP-2). In some embodiments, the immune-cell targeting polypeptide is capable of targeting the ENP to one or more immune cells each selected from the group comprising: a T-cell, a B-cell, a macrophage, a neutrophil, a dendritic cell, optionally a follicular dendritic cell, an innate lymphoid cell, a mast cell, an eosinophil, a basophil, a megakaryocyte, or a natural killer cell.

[0173] In some embodiments, the cell surface domain and the chimeric protein transmembrane domain are separated by a linker, wherein the linker: is a flexible linker, a rigid linker, or a hybrid linker; is hydrophilic or hydrophobic; is between 1 and 250 amino acids (e.g.,1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55,56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81,82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 150, 200, 250, or a number between any two of these values, amino acids in length); comprises one or more flexible amino acid residues, optionally about 1 to about 250 flexible amino acid residues, further optionally the flexible amino acid residues comprise glycine, serine, or a combination thereof. In some embodiments, the linker comprises 3 repeating amino acid subunits or more.

[0174] The cell surface domain can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to SEQ ID NO: 128 or SEQ ID NO: 130.

[0175] As described herein, the recombinant protein and the chimeric protein can have the same or similar transmembrane and / or cytoplasmic domains, thereby, co-localizing the recombinant protein and the chimeric protein at the plasma membrane. The cytoplasmic domain and / or transmembrane domain of the recombinant protein can comprise an amino acid sequence having at least 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to the sequence of the cytoplasmic domain and / or the transmembrane domain of the chimeric protein. In some embodiments, the endogenous transmembrane domain (and optionally, endogenous cytoplasmic tail) of the antigenic polypeptide is used as transmembrane domain sequence for the chimeric protein. In some embodiments, a transmembrane domain (and, optionally, cytoplasmic tail) heterologous to the antigenic polypeptide is used as the transmembrane domain for both the recombinant protein and the chimeric peptide. The transmembrane domain and the cytoplasmic domain of the recombinant protein and the transmembrane domain and the cytoplasmic domain of the chimeric protein each can comprise an amino acid sequence having at least 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 129 (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values). The recombinant protein and / or the chimeric protein can further comprise an ERD. In some embodiments, the ERD is at the C-terminus of the recombinant protein and / or the chimeric protein.

[0176] The chimeric protein can comprise an amino acid sequence having at least65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) sequence identity to the sequence of SEQ ID NO: 124, SEQ ID NO: 125, or SEQ ID NO: 126. The AP of the recombinant protein can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to SEQ ID NO: 40 or SEQ ID NO: 47.

[0177] In some embodiments, one or more of (i) the polynucleotide encoding the recombinant protein or the n polynucleotides each encoding an nth recombinant protein, and (ii) the polynucleotide encoding the chimeric protein or the z polynucleotides each encoding a zth chimeric protein, are present in a same or different nucleic acid molecule. The amount of (i) the polynucleotide encoding the recombinant protein or the n polynucleotides each encoding an nth recombinant protein; and (ii) the polynucleotide encoding the chimeric protein or the z polynucleotides each encoding a zth chimeric protein can vary. In some embodiments, the amount of (i) the polynucleotide encoding the recombinant protein or the n polynucleotides each encoding an nth recombinant protein; and (ii) the polynucleotide encoding the chimeric protein or the z polynucleotides each encoding a zth chimeric protein, are present in the composition at a molar ratioof about 10:1, 4:1, or 1:1 (e.g., 1:1, 1.1:1, 1.2:1, 1.3:1, 1.4:1, 1.5:1, 1.6:1, 1.7:1, 1.8:1, 1.9:1, 2:1, 2.5:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, 20:1, 21:1, 22:1, 23:1, 24:1, 25:1, 26:1, 27:1, 28:1, 29:1, 30:1, 31:1, 32:1, 33:1, 34:1, 35:1, 36:1,37:1, 38:1, 39:1, 40:1, 41:1, 42:1, 43:1, 44:1, 45:1, 46:1, 47:1, 48:1, 49:1, 50:1, 51:1, 52:1, 53:1,54:1, 55:1, 56:1, 57:1, 58:1, 59:1, 60:1, 61:1, 62:1, 63:1, 64:1, 65:1, 66:1, 67:1, 68:1, 69:1, 70:1,71:1, 72:1, 73:1, 74:1, 75:1, 76:1, 77:1, 78:1, 79:1, 80:1, 81:1, 82:1, 83:1, 84:1, 85:1, 86:1, 87:1,88:1, 89:1, 90:1, 91:1, 92:1, 93:1, 94:1, 95:1, 96:1, 97:1, 98:1, 99:1, 100:1, or a number or a range between any of these values). In some embodiments, the polynucleotide encoding the recombinant protein and the polynucleotide encoding the chimeric protein are present in the same nucleic acid molecule.

[0178] In some embodiments, the cell expressing the recombinant protein or the plurality of recombinant proteins and the chimeric protein or plurality of chimeric proteins exhibits at least a 2-fold increase (e.g., 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 20-fold, 30-fold, 40-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold, 100-fold, or a number or a range between any of these values) in AP expression at the surface of the cell, relativeto a cell expressing a fusion protein or a plurality of fusion proteins comprising an AP and an ERD and does not express the chimeric protein. The production of ENPs from the cell expressing the recombinant protein or the plurality of recombinant proteins and the chimeric protein or plurality of chimeric proteins can be increased by at least 2-fold (e.g., 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 20-fold, 30-fold, 40-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90- fold, 100-fold, or a number or a range between any of these values), relative to a cell expressing a fusion protein or a plurality of fusion proteins comprising an AP and an ERD and does not express the adapter fusion protein.

[0179] In some embodiments, the nucleic acid composition further comprises a polynucleotide comprising or encoding a tetherin inhibitor. In some embodiments, the tetherin inhibitor is capable of modulating expression, concentration, localization, stability, and / or activity of tetherin. In some embodiments, the tetherin inhibitor comprises a dsRNA, an siRNA, an shRNA, a pre-miRNA, a pri-miRNA, a miRNA, an stRNA, an IncRNA, a piRNA, a snoRNA, or a protein. In some embodiments, one or more of (i) the polynucleotide comprising or encoding the tetherin inhibitor, (ii) the polynucleotide encoding the recombinant protein or the n polynucleotides each encoding an nth recombinant protein, and (iii) the polynucleotide encoding the chimeric protein or the z polynucleotides each encoding a zth chimeric protein, are present in a same or different nucleic acid molecule. In some embodiments, the amount of (i) the polynucleotide comprising or encoding the tetherin inhibitor; and (ii) the polynucleotide encoding the recombinant protein or the n polynucleotides each encoding an nth recombinant protein and / or the polynucleotide encoding the chimeric protein or the z polynucleotides each encoding a zth chimeric protein, are present in the composition at a molar ratio of about 1:1, 1 :5 or 1 :25. In some embodiments, the amount of (i) the polynucleotide comprising or encoding the tetherin inhibitor; and (ii) the polynucleotide encoding the recombinant protein or the n polynucleotides each encoding an nth recombinant protein and / or the polynucleotide encoding the chimeric protein or the z polynucleotides each encoding a zth chimeric protein, are present in the composition at a molar ratio of about e.g., 1:1, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9, 1:2, 1:2.5, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, 1:20, 1:21,1:22, 1:23, 1:24, 1:25, 1:26, 1:27, 1:28, 1:29, 1:30, 1:31, 1:32, 1:33, 1:34, 1:35, 1:36, 1:37, 1:38,1:39, 1:40, 1:41, 1:42, 1:43, 1:44, 1:45, 1:46, 1:47, 1:48, 1:49, 1:50, 1:51, 1:52, 1:53, 1:54, 1:55,1:56, 1:57, 1:58, 1:59, 1:60, 1:61, 1:62, 1:63, 1:64, 1:65, 1:66, 1:67, 1:68, 1:69, 1:70, 1:71, 1:72,1:73, 1:74, 1:75, 1:76, 1:77, 1:78, 1:79, 1:80, 1:81, 1:82, 1:83, 1:84, 1:85, 1:86, 1:87, 1:88, 1:89,1:90, 1:91, 1:92, 1:93, 1:94, 1:95, 1:96, 1:97, 1:98, 1:99, 1:100 or a number or a range between any of these values. In some embodiments, the polynucleotide comprising or encoding the tetherin inhibitor and the polynucleotide encoding the recombinant protein are present in the same nucleicacid. In some embodiments, the polynucleotide comprising or encoding the tetherin inhibitor and the polynucleotide encoding the chimeric protein are present in the same nucleic acid. For example, the polynucleotide encoding the tetherin inhibitor and the polynucleotide encoding the recombinant protein or the chimeric protein can be operably linked to a tandem gene expression element (e.g., an internal ribosomal entry site (IRES), foot-and-mouth disease virus 2A peptide (F2A), equine rhinitis A virus 2A peptide (E2A), porcine teschovirus 2A peptide (P2A) or Thosea asigna virus 2A peptide (T2A), or any combination thereof).

[0180] In some embodiments, the tetherin inhibitor comprises or is derived from a viral protein, optionally the virus is HIV-1, HIV-2, SIV, Ebola virus, KSHV, SARS CoV, or SARS-CoV-2. In some embodiments, the tetherin inhibitor comprises HIV-1 Vpu protein, KSHV K5 protein, SARS-CoV-2 ORF7a, HIV-2 Env, Ebola GP, SIV Env, SIV Vpu, SIV Nef, or any portions, variants or derivatives thereof. In some embodiments, the tetherin inhibitor comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 132-134. In some embodiments, the tetherin inhibitor comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) sequence identity to the sequence of SEQ ID NO: 132. In some embodiments, presence or expression of the tetherin inhibitor in the cell results in an increase in ENP production by the cell by at least 2-fold (e.g., 2-fold, 3-fold, 4-fold, 5-fold, 6- fold, 7-fold, 8-fold, 9-fold, 10-fold, 20-fold, 30-fold, 40-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold, 100-fold, or a number or a range between any of these values), relative to a cell that does not comprise or express the tetherin inhibitor.Endosomal Sorting Complex Required for Transport (ESCRT)-recruiting domain (ERD)

[0181] In some embodiments, an adapter fusion protein of the disclosure comprises an endosomal sorting complex required for transport (ESCRT)-recruiting domain (ERD). In some embodiments, a recombinant protein and / or chimeric protein of the disclosure can further comprise an endosomal sorting complex required for transport (ESCRT)-recruiting domain (ERD). In some embodiments, the recombinant protein comprises an ERD (e.g., at the C-terminal or cytosolic portion of the protein). In some embodiments, the chimeric protein comprises the ERD (e.g., at the C-terminal or cytosolic portion of the protein). The ERD is capable of recruitingone or more ESCRT proteins, thereby inducing production and secretion of ENPs from the cell.

[0182] The ENPs can comprise a lipid bilayer. In some embodiments, the ENPs can comprise a lipid bilayer derived from the cell from which the ENP was secreted. The ERD can be located at the C-terminus of the dimerization fusion protein, the adapter fusion protein, the recombinant protein and / or the chimeric protein. In some embodiments, the ERD is capable of interacting with the ESCRT proteins TSG101, NEDD4, and / or ALIX.

[0183] In some embodiments, the ERD comprises or is derived from a nonhuman protein. In some embodiments, the ERD comprises or is derived from a human protein. In some embodiments, the ERD comprises or is derived from a nonmammalian protein. In some embodiments, the ERD comprises or is derived from a chicken protein, a mouse protein, a lizard protein, a reptile protein, a hamster protein, or a goldfish protein. In some embodiments, the ERD comprises or is derived from the ESCRT and ALIX binding region (EABR) of the human CEP55 protein. In some embodiments, the ERD comprises or is derived from residues 170-213 of the human CEP55 protein. In some embodiments, the ERD comprises or is derived from Syntenin-1, rat Galectin-3 (rGalectin-3), Hrs, and / or CD2AP. In some embodiments, the ERD comprises or is derived from a viral protein. In some embodiments, the ERD comprises or is derived from a fragment of a viral protein. In some embodiments, the ERD comprises or is derived from a retroviral protein, herpes simplex viral protein, vaccinia viral protein, hepadnaviral protein, togaviral protein, flaviviral protein, arenaviral protein, coronaviral protein, orthomyxoviral protein, paramyxoviral protein, bunyaviral protein, bornaviral protein, rhabdoviral protein or filoviral protein. In some embodiments, the ERD comprises or is derived from a Gag protein. In some embodiments, the ERD comprises or is derived from EIAV, HTLV-1, MLV, or MPMV. In some embodiments, the ERD comprises or is derived from EIAV p9, SIV p6 and / or HIV-1 p6; and / or an Ebola protein. In some embodiments, the ERD comprises or is derived from EBOV VP40.

[0184] In some embodiments, the ERD comprises or is derived from at least a portion of CEP55 protein. The ERD can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to the amino acid sequence of SEQ ID NO: 2.

[0185] The ERD can comprise one or more TSG101 -binding motifs, one or more ALIX-binding motifs, one or more Nedd4-recruiting motifs, or any combination thereof. In some embodiments, any two of the one or more TSG101 -binding motifs, the one or more ALIX-bindingmotifs, or the one or more Nedd4-recruiting motifs are the same or different. In some embodiments, the ERD comprises or is derived from HIV-1 p6 protein or SIV p6 protein. In some embodiments, the ERD comprises or is derived from the p6 protein of HIV-1 isolate ETH2220. The ERD can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to an amino acid sequence selected from the group consisting of SEQ ID NOs: 5-37.

[0186] In some embodiments, the ERD comprises or is derived from a non-human galectin protein. In some embodiments, the ERD comprises or is derived from a rat galectin protein. In some embodiments, the ERD comprising or derived from a non-human galectin protein further comprises a viral-derived ALIX-recruiting motif. The ERD can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to an amino acid sequence selected from the group consisting of SEQ ID NOs: 38-39. Provided in Table 1 are exemplary ERDs that can be used, e.g., in the adapter fusion proteins, the recombinant proteins, and / or the chimeric proteins of the disclosure.Table 1 : Exemplary ERDs

[0187] The dimerization fusion protein, the adapter fusion protein, the recombinant protein, and / or the chimeric protein can comprise an endocytosis-preventing motif (EPM) capable of preventing endocytosis of the dimerization fusion protein, the adapter fusion protein, the recombinant protein, and / or the chimeric protein. In some embodiments, the EPM: tethers the dimerization fusion protein, the adapter fusion protein, the recombinant protein, and / or the chimeric protein to the cytoskeleton, thereby preventing localization to coated pits and endocytosis; enhances ENP assembly, ENP production, and / or ENP secretion; and / or prevents endocytosis of the dimerization fusion protein, the adapter fusion protein, the recombinant protein, and / or the chimeric protein, thereby extending the time the dimerization fusion protein, the adapter fusion protein, the recombinant protein, and / or the chimeric protein remains at the plasma membrane to interact with ESCRT proteins.

[0188] In some embodiments, the EPM: increases the abundance and / or density of dimerization fusion proteins, adapter fusion proteins, recombinant proteins, and / or chimeric proteins on and / or in the ENP by at least about 2-fold (e.g., 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 20-fold, 30-fold, 40-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90- fold, 100-fold, or a number or a range between any of these values) as compared to an ENP comprising a dimerization fusion protein, an adapter fusion protein, a recombinant protein, and / or a chimeric protein that does not comprise the EPM; and / or increases the number of ENPs secreted by a cell by at least about 2-fold (e.g., 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 20-fold, 30-fold, 40-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold, 100-fold, or a number or a range between any of these values) as compared to a cell expressing a dimerization fusion protein, an adapter fusion protein, a recombinant protein, and / or a chimeric protein that does not comprise the EPM.

[0189] In some embodiments, the EPM comprises or is derived from a portion of murine low-affinity gamma Fc region receptor II isoform FcRII-Bl. The EPM can comprise all or a portion of the cytoplasmic tail of FcRII-Bl. The EPM can comprise an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity (e.g., at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or a range between any two of these values) to the amino acid sequence of SEQ ID NO: 1.

[0190] In some embodiments, the dimerization fusion protein, the adapter fusion protein, the recombinant protein, and / or the chimeric protein does not comprise an endocytosis- preventing motif (EPM).Pathogenic Antigens

[0191] The AP can comprise or can be derived from the full-length surface protein of an infectious agent. The disease or disorder can be an infectious disease or disorder caused by an infectious agent, the AP can comprise or can be derived from an antigenic protein of said infectious agent, and the antigenic protein of said infectious agent can be a pathogenic antigen. In some embodiments, the pathogenic antigen is selected from the group comprising: Outer membrane protein A OmpA, biofilm associated protein Bap, transport protein MucK (Acinelobacler baumannii, Acinetobacter infections)); variable surface glycoprotein VSG, microtubule-associated protein MAPP15, trans-sialidase TSA (Trypanosoma brucei. African sleeping sickness (African trypanosomiasis)); HIV p24 antigen, HIV envelope proteins (Gpl20, Gp41, Gpl60), polyprotein GAG, negative factor protein Nef, trans-activator of transcription Tat (HIV (Human immunodeficiency virus), AIDS (Acquired immunodeficiency syndrome)); galactose-inhibitable adherence protein GIAP, 29 kDa antigen Eh29, Gal / GalNAc lectin, protein CRT, 125 kDa immunodominant antigen, protein M17, adhesin ADH112, protein STIRP (Entamoeba histolytica, Amoebiasis); Major surface proteins 1-5 (MSPla, MSPlb, MSP2, MSP3, MSP4, MSP5), type IV secretion system proteins (VirB2, VirB7, VirBl l, VirD4) (Anaplasma genus, Anaplasmosis); protective Antigen PA, edema factor EF, lethal factor LF, the S-layer homology proteins SLH (Bacillus anlhracis. Anthrax); acranolysin, phospholipase D, collagen-binding protein CbpA (Arcanobacterium haemolyticum, Arcanobacterium haemolyticum infection); nucleocapsid protein NP, glycoprotein precursor GPC, glycoprotein GP1, glycoprotein GP2 (Junin virus, Argentine hemorrhagic fever); chitin-protein layer proteins, 14 kDa surface antigen A14, major sperm protein MSP, MSP polymerization-organizing protein MPOP, MSP fiber protein 2 MFP2, MSP polymerization-activating kinase MPAK, ABA- 1 -like protein ALB, protein ABA-1, cuticulin CUT-1 (Ascaris himbricoides, Ascariasis); 41 kDaallergen Asp vl3, allergen Asp f3, major coni dial surface protein rodlet A, protease Pep Ip, GPI- anchored protein Gel Ip, GPI-anchored protein Crap (Aspergillus genus, Aspergillosis); family VP26 protein, VP29 protein (Astroviridae, Astrovirus infection); Rhoptry-associated protein 1 RAP-1, merozoite surface antigens MSA-1, MSA-2 (al, a2, c), 12D3, 1105, 21134, P29, variant erythrocyte surface antigen VESA1, Apical Membrane Antigen 1 AMA-1 (Babesia genus, Babesiosis); hemolysin, enterotoxin C, PX01-51, glycolate oxidase, ABC-transporter, penicillin- binding protein, zinc transporter family protein, pseudouridine synthase Rsu, plasmid replication protein RepX, oligoendopeptidase F, prophage membrane protein, protein HemK, flagellar antigen H, 28.5-kDa cell surface antigen (Bacillus cereus, Bacillus cereus infection); large T antigen LT, small T antigen, capsid protein VP1, capsid protein VP2 (BK virus, BK virus infection); 29 kDa-protein, caspase-3 -like antigens, glycoproteins (Blastocystis hominis, Blastocystis hominis infection ,' yeast surface adhesin WI-1 (Blastomyces dermatitidis, Blastomycosis); nucleoprotein N, polymerase L, matrix protein Z, glycoprotein GP (Machupo virus, Bolivian hemorrhagic fever); outer surface protein A OspA, outer surface protein OspB, outer surface protein OspC, decorin binding protein A DbpA, decorin binding protein B DbpB, flagellar filament 41 kDa core protein Fla, basic membrane protein A precursor Bmp A (Immunodominant antigen P39), outer surface 22 kDa lipoprotein precursor (antigen IPLA7), variable surface lipoprotein vlsE (Borrelia genus, Borrelia infection); Botulinum neurotoxins BoNT / Al, BoNT / A2, BoNT / A3, BoNT / B, BoNT / C, BoNT / D, BoNT / E, BoNT / F, BoNT / G, recombinant botulinum toxin F He domain FHc (Clostridium botulinum, Botulism (and Infant botulism)); nucleocapsid, glycoprotein precursor (Sabia virus, Brazilian hemorrhagic fever); copper / Zinc superoxide dismutase SodC, bacterioferritin Bfr, 50S ribosomal protein RplL, OmpA-like transmembrane domain-containing protein 0mp31, immunogenic 39-kDa protein M5 P39, zinc ABC transporter periplasmic zinc-binding protein znuA, periplasmic immunogenic protein Bp26, 30S ribosomal protein S12 RpsL, glyceraldehyde-3 -phosphate dehydrogenase Gap, 25 kDa outer-membrane immunogenic protein precursor Omp25, invasion protein B lalB, trigger factor Tig, molecular chaperone DnaK, putative peptidyl-prolyl cis-trans isomerase SurA, lipoprotein 0mpl9, outer membrane protein MotY 0mpl6, conserved outer membrane protein D15, malate dehydrogenase Mdh, component of the Type-IV secretion system (TOSS) VirJ, lipoprotein of unknown function BAB 1 0187 (Brucella genus, Brucellosis); members of the ABC transporter family (LoIC, OppA, and PotF), putative lipoprotein releasing system transmembrane protein LoICZE, flagellin FliC, Burkholderia intracellular motility A BimA, bacterial Elongation factor-Tu EF-Tu, 17 kDa OmpA-like protein, boaA coding protein, boaB coding protein (Burkholderia cepacia and other Burkholderia species, Burkholderia infection); mycolyl- transferase Ag85A, heat-shock protein Hsp65, protein TB10.4, 19 kDa antigen, protein PstS3,heat-shock protein Hsp70 (Mycobacterium ulcerans, Buruli ulcer); norovirus major and minor viral capsid proteins VP1 and VP2, genome polyprotein, Sapoviurus capsid protein VP1, protein Vp3, genome polyprotein (Caliciviridae family, Calicivirus infection (Norovirus and Sapovirus)); major outer membrane protein PorA, flagellin FlaA, surface antigen CjaA, fibronectin binding protein CadF, aspartate / glutamate-binding ABC transporter protein PeblA, protein FspAl, protein FspA2 (Campylobacter genus, Campylobacteriosis); glycolytic enzyme enolase, secreted aspartyl proteinases SAP1-10, glycophosphatidylinositol (GPI)-linked cell wall protein, protein Hyrl, complement receptor 3-related protein CR3-RP, adhesin Als3p, heat shock protein 90 kDa hsp90, cell surface hydrophobicity protein CSH (usually Candida albicans and other Candida species, Candidiasis); 17-kDa antigen, protein P26, trimeric autotransporter adhesins TAAs, Bartonella adhesin A BadA, variably expressed outer-membrane proteins Vomps, protein Pap3, protein HbpA, envelope-associated protease HtrA, protein OMP89, protein GroEL, protein LalB, protein OMP43, dihydrolipoamide succinyltransferase SucB (Bartonella henselae, Catscratch disease); amastigote surface protein-2, amastigote-specific surface protein SSP4, cruzipain, trans-sialidase TS, trypomastigote surface glycoprotein TSA-1, complement regulatory protein CRP-10, protein G4, protein G2, paraxonemal rod protein PAR2, paraflagellar rod component Part, mucin-Associated Surface Proteins MPSP (Trypanosoma cruzi, Chagas Disease (American trypanosomiasis)); envelope glycoproteins (gB, gC, gE, gH, gl, gK, gL) (Varicella zoster virus (VZV), Chickenpox); major outer membrane protein MOMP, probable outer membrane protein PMPC, outer membrane complex protein B OmcB, heat shock proteins Hsp60 HSP10, protein IncA, proteins from the type III secretion system, ribonucleotide reductase small chain protein NrdB, plasmid protein Pgp3, chlamydial outer protein N CopN, antigen CT521, antigen CT425, antigen CT043, antigen TC0052, antigen TC0189, antigen TC0582, antigen TC0660, antigen TC0726, antigen TC0816, antigen TC0828 (Chlamydia trachomatis, Chlamydia , low calcium response protein E LCrE, chlamydial outer protein N CopN, serine / threonine-protein kinase PknD, acyl-carrier-protein S-malonyltransferase FabD, singlestranded DNA-binding protein Ssb, major outer membrane protein MOMP, outer membrane protein 2 0mp2, polymorphic membrane protein family (Pmpl, Pmp2, Pmp3, Pmp4, Pmp5, Pmp6, Pmp7, Pmp8, Pmp9, PmplO, Pmpl l, Pmpl2, Pmpl3, Pmpl4, Pmpl5, Pmpl6, Pmpl7, Pmpl 8, Pmpl 9, Pmp20, Pmp21) (Chlamydophila pneumoniae, Chlamydophila pneumoniae infection); cholera toxin B CTB, toxin coregulated pilin A TcpA, toxin coregulated pilin TcpF, toxin co-regulated pilus biosynthesis ptrotein F TcpF, cholera enterotoxin subunit A, cholera enterotoxin subunit B, Heat-stable enterotoxin ST, mannose-sensitive hemagglutinin MSHA, outer membrane protein U Porin ompU, Poring B protein, polymorphic membrane protein-D (Vibrio cholerae, Cholera); propionyl-CoA carboxylase PCC, 14-3-3 protein,-n-prohibitin, cysteine proteases, glutathione transferases, gelsolin, cathepsin L proteinase CatL, Tegumental Protein 20.8 kDa TP20.8, tegumental protein 31.8 kDa TP31.8, lysophosphatidic acid phosphatase LPAP, (Clonorchis sinensis, Clonorchiasis); surface layer proteins SLPs, glutamate dehydrogenase antigen GDH, toxin A, toxin B, cysteine protease Cwp84, cysteine protease Cwpl3, cysteine protease Cwpl9, Cell Wall Protein CwpV, flagellar protein FliC, flagellar protein FliD (Clostridium difficile, Clostridium difficile infection); rhinoviruses: capsid proteins VP1, VP2, VP3, VP4; coronaviruses: spike proteins S, envelope proteins E, membrane proteins M, nucleocapsid proteins N (usually rhinoviruses and coronaviruses, Common cold (Acute viral rhinopharyngitis; Acute coryza)); prion protein Prp (CJD prion, Creutzfeldt-Jakob disease (CJD)); envelope protein Gc, envelope protein Gn, nucleocapsid proteins (Crimean-Congo hemorrhagic fever virus, Crimean-Congo hemorrhagic fever (CCHF)); virulence-associated DEAD-box RNA helicase VAD1, galactoxylomannan-protein GalXM, glucuronoxylomannan GXM, mannoprotein MP (Cryptococcus neoformans, Cryptococcosis); acidic ribosomal protein P2 CpP2, mucin antigens Mucl, Muc2, Muc3 Muc4, Muc5, Muc6, Muc7, surface adherence protein CP20, surface adherence protein CP23, surface protein CP 12, surface protein CP21, surface protein CP40, surface protein CP60, surface protein CP 15, surface-associated glycopeptides gp40, surface- associated glycopeptides gpl5, oocyst wall protein AB, profilin PRF, apyrase (Cryptosporidium genus, Cryptosporidiosis); fatty acid and retinol binding protein- 1 FAR-1, tissue inhibitor of metalloproteinase TIMP (TMP), cysteine proteinase ACEY-1, cysteine proteinase ACCP-1, surface antigen Ac- 16, secreted protein 2 ASP-2, metalloprotease 1 MTP-1, aspartyl protease inhibitor API-1, surface-associated antigen SAA-1, adult-specific secreted factor Xa serine protease inhibitor anticoagulant AP, cathepsin D-like aspartic protease ARR-1 (usually Ancylostoma braziliense,' multiple other parasites, Cutaneous larva migrans (CLM)); cathepsin L-like proteases, 53 / 25-kDa antigen, 8 kDa family members, cysticercus protein with a marginal trypsin-like activity TsAg5, oncosphere protein TSOL18, oncosphere protein TSOL45- 1A, lactate dehydrogenase A LDHA, lactate dehydrogenase B LDHB (Taenia solium, Cysticercosis); pp65 antigen, membrane protein ppi 5, capsid-proximal tegument protein ppi 50, protein M45, DNA polymerase UL54, helicase ULI 05, glycoprotein gM, glycoprotein gN, glycoprotein H, glycoprotein B gB, protein UL83, protein UL94, protein UL99 (Cytomegalovirus (CMV), Cytomegalovirus infection); capsid protein C, premembrane protein prM, membrane protein M, envelope protein E (domain I, domain II, domain II), protein NS1, protein NS2A, protein NS2B, protein NS3, protein NS4A, protein 2K, protein NS4B, protein NS5 (Dengue viruses (DEN-1, DEN-2, DEN-3 and DEN-4)-Flaviviruses, Dengue fever); 39 kDa protein (Dientamoeba fragilis, Dientamoebiasis); diphtheria toxin precursor Tox, diphtheria toxin DT, pilin-specific sortase SrtA, shaft pilin protein SpaA, tip pilin protein SpaC, minor pilin proteinSpaB, surface-associated protein DIP 1281 (Corynebacterium diphlheriae, Diphtheria); glycoprotein GP, nucleoprotein NP, minor matrix protein VP24, major matrix protein VP40, transcription activator VP30, polymerase cofactor VP35, RNA polymerase L (Ebolavirus (EBOV), Ebola hemorrhagic fever); prion protein (vCJD prion, Variant Creutzfeldt-Jakob disease (vCJD, nvCJD)); UvrABC system protein B, protein Flpl, protein Flp2, protein Flp3, protein TadA, hemoglobin receptor HgbA, outer membrane protein TdhA, protein CpsRA, regulator CpxR, protein SapA, 18 kDa antigen, outer membrane protein NcaA, protein LspA, protein LspAl, protein LspA2, protein LspB, outer membrane component DsrA, lectin DItA, lipoprotein Hip, major outer membrane protein OMP, outer membrane protein 0mpA2 (Haemophilus ducreyi, Chancroid); aspartyl protease 1 Pepl, phospholipase B PLB, alpha-mannosidase 1 AMN1, glucanosyltransferase GEL1, urease URE, peroxisomal matrix protein Pmpl, proline-rich antigen Pra, humal T-cell reative protein TcrP (Coccidioides immitis and Coccidioides posadasii, Coccidioidomycosis); allergen Tri r 2, heat shock protein 60 Hsp60, fungal actin Act, antigen Tri r2, antigen Tri r4, antigen Tri tl, protein IV, glycerol-3 -phosphate dehydrogenase Gpdl, osmosensor HwSholA, osmosensor HwSholB, histidine kinase HwHhk7B, allergen Mala s 1, allergen Mala s 11, thioredoxin Trx Mala s 13, allergen Mala f, allergen Mala s (usually Trichophyton spp, Epidermophyton spp., Malassezia spp., Hortaea w erneckii,Dermatophytosis); protein EG95, protein EG10, protein EG18, protein EgA31, protein EM18, antigen EPCI, antigen B, antigen 5, protein P29, protein 14-3-3, 8-kDa protein, myophilin, heat shock protein 20 HSP20, glycoprotein GP-89, fatty acid binding protein FAPB (Echinococcus genus, Echinococcosis); major surface protein 2 MSP2, major surface protein 4 MSP4, MSP variant SGV1, MSP variant SGV2, outer membrane protein OMP, outer membrane protein 19 OMP- 19, major antigenic protein MAPI, major antigenic protein MAP 1-2, major antigenic protein MAP1B, major antigenic protein MAPI-3, Erum2510 coding protein, protein GroEL, protein GroES, 30-kDA major outer membrane proteins, GE 100-kDa protein, GE 130- kDa protein, GE 160-kDa protein (Ehrlichia genus, Ehrlichiosis); secreted antigen SagA, sagA- like proteins Sal A and SalB, collagen adhesin Scm, surface proteins Fmsl (EbpA(fm), Fms5 (EbpB(fm), Fms9 (EpbC(fm) and FmslO, protein EbpC(fm), 96 kDa immunoprotective glycoprotein G1 (Enterococcus genus, Enterococcus infection); genome polyprotein, polymerase 3D, viral capsid protein VP1, viral capsid protein VP2, viral capsid protein VP3, viral capsid protein VP4, protease 2 A, protease 3C (Enterovirus genus, Enterovirus infection); outer membrane proteins OM, 60 kDa outer membrane protein, cell surface antigen OmpA, cell surface antigen OmpB (sca5), 134 kDa outer membrane protein, 31 kDa outer membrane protein, 29.5 kDa outer membrane protein, cell surface protein SCA4, cell surface protein Adri (RP827), cell surface protein Adr2 (RP828), cell surface protein SCA1, Invasion protein invA, cell divisionprotein fts, secretion proteins sec Ofamily, virulence proteins virB, tlyA, tlyC, parvulin-like protein Pip, preprotein translocase SecA, 120-kDa surface protein antigen SPA, 138 kD complex antigen, major 100-kD protein (protein I), intracytoplasmic protein D, protective surface protein antigen SPA (Rickettsia prowazekii, Epidemic typhus); Epstein-Barr nuclear antigens (EBNA-1, EBNA-2, EBNA-3A, EBNA-3B, EBNA-3C, EBNA-leader protein (EBNA-LP)), latent membrane proteins (LMP-1, LMP-2A, LMP-2B), early antigen EBV-EA, membrane antigen EBV-MA, viral capsid antigen EBV-VCA, alkaline nuclease EBV-AN, glycoprotein glycoprotein gp350, glycoprotein gpl lO, glycoprotein gp42, glycoprotein gHgL, glycoprotein gB (Epstein- Barr Virus (EBV), Epstein-Barr Virus Infectious Mononucleosis); cpasid protein VP2, capsid protein VP1, major protein NS1 (Parvovirus B19, Erythema infectiosum (Fifth disease)); pp65 antigen, glycoprotein 105, major capsid protein, envelope glycoprotein H, protein U51 (Human herpesvirus 6 (HHV-6) and Human herpesvirus 7 (HHV-7), Exanthem subitum); thioredoxin- glutathione reductase TGR, cathepsins LI and L2, Kunitz-type protein KTM, leucine aminopeptidase LAP, cysteine proteinase Fast, saposin-like protein-2 SAP-2, thioredoxin peroxidases TPx, Prx-1, Prx-2, cathepsin I cysteine proteinase CL3, protease cathepsin L CL1, phosphoglycerate kinase PGK, 27-kDa secretory protein, 60 kDa protein HSP35alpha, glutathione transferase GST, 28.5 kDa tegumental antigen 28.5 kDa TA, cathepsin B3 protease CatB3, Type I cystatin stefin-1, cathepsin L5, cathepsin Llg and cathepsin B, fatty acid binding protein FABP, leucine aminopeptidases LAP (Fasciola hepatica and Fasciola gigantica, Fasciolosis); prion protein (FFI prion, Fatal familial insomnia (FFI)); venom allergen homolog-like protein VAL-1, abundant larval transcript ALT-1, abundant larval transcript ALT-2, thioredoxin peroxidase TPX, vespid allergen homologue VAH, thiordoxin peroxidase 2 TPX-2, antigenic protein SXP (peptides N, Nl, N2, and N3), activation associated protein- 1 ASP-1, Thioredoxin TRX, transglutaminase BmTGA, glutathione-S-transferases GST, myosin, vespid allergen homologue VAH, 175 kDa collagenase, glyceraldehyde-3 -phosphate dehydrogenase GAPDH, cuticular collagen Col-4, secreted larval acidic proteins SLAPs, chitinase CHI-1, maltose binding protein MBP, glycolytic enzyme fructose- 1,6-bisphosphate aldolase Fba, tropomyosin TMY-1, nematode specific gene product OvB20, onchocystatin CPI-2, Cox-2 (Filarioidea superfamily, Filariasis); phospholipase C PLC, heat-labile enterotoxin B, Iota toxin component lb, protein CPE1281 pyruvate ferredoxin oxidoreductase, elongation factor G EF-G, perfringolysin 0 Pfo, glyceraldehyde-3 -phosphate dehydrogenase GapC, Fructose-bisphosphate aldolase Alf2, Clostridium perfringens enterotoxin CPE, alpha toxin AT, alpha toxoid ATd, epsilon-toxoid ETd, protein HP, large cytotoxin TpeL, endo-beta-N-acetylglucosaminidase Naglu, phosphoglyceromutase Pgm (Clostridium perfringens, Food poisoning by Clostridium perfringens),' leukotoxin IktA, adhesion FadA, outer membrane protein RadD, high-molecular weight arginine-binding protein(Fusobacterium genus, Fusobacterium infection); phospholipase C PLC, heat-labile enterotoxin B, Iota toxin component lb, protein CPE1281, pyruvate ferredoxin oxidoreductase, elongation factor GEF-G, perfringolysin 0 Pfo, glyceraldehyde-3 -phosphate dehydrogenase GapC, fructosebisphosphate aldolase Alf2, Clostridium perfringens enterotoxin CPE, alpha toxin AT, alpha toxoid ATd, epsilon-toxoid ETd, protein HP, large cytotoxin TpeL, endo-beta-N- acetylglucosaminidase Naglu, phosphoglyceromutase Pgm (usually Clostridium perfringens,' other Clostridium species, Gas gangrene (Clostridial myonecrosis)); lipase A, lipase B, peroxidase Decl (Geotrichum candidum. Geotrichosis); prion protein (GSS prion, Gerstmann- Straussler-Scheinker syndrome (GSS)); cyst wall proteins CWP1, CWP2, CWP3, variant surface protein VSP, VSP1, VSP2, VSP3, VSP4, VSP5, VSP6, 56 kDa antigen, pyruvate ferredoxin oxidoreductase PF OR, alcohol dehydrogenase E ADHE, alpha-giardin, alpha8-giardin, alpha 1- guiardin, beta-giardin, cystein proteases, glutathione-S-transferase GST, arginine deiminase ADI, fructose- 1,6-bisphosphat aldolase FBA, Giardia trophozoite antigens GTA (GTA1, GTA2), ornithine carboxyl transferase OCT, striated fiber-asseblin-like protein SALP, uridine phosphoryl- like protein UPL, alpha-tubulin, beta-tubulin (Giardia inleslinalis. Giardiasis); members of the ABC transporter family (LoIC, OppA, and PotF), putative lipoprotein releasing system transmembrane protein LoICZE, flagellin FliC, Burkholderia intracellular motility A BimA, bacterial Elongation factor-Tu EF-Tu, 17 kDa OmpA-like protein, boaA coding protein (Burkholderia mallei, Glanders); cyclophilin CyP, 24 kDa third-stage larvae protein GS24, excretion- secretion products ESPs (40, 80, 120 and 208 kDa) (Gnathostoma spinigerum and Gnathostoma hispidum, Gnathostomiasis); pilin proteins, minor pilin-associated subunit pilC, major pilin subunit and variants pilE, pilS, phase variation protein porA, Porin B PorB, protein TraD, Neisserial outer membrane antigen H.8, 70 kDa antigen, major outer membrane protein PI, outer membrane proteins PIA and PIB, W antigen, surface protein A NspA, transferrin binding protein TbpA, transferrin binding protein TbpB PBP2, mtrR coding protein, ponA coding protein, membrane permease FbpBC, FbpABC protein system, LbpAB proteins, outer membrane protein Opa, outer membrane transporter FetA, iron-repressed regulator MpeR (Neisseria gonorrhoeae. Gonorrhea); outer membrane protein A OmpA, outer membrane protein C OmpC, outer membrane protein K17 0mpK17 (Klebsiella granulomalis. Granuloma inguinale (Donovanosis)); fibronectin-binding protein Sfb, fibronectin / fibrinogen-binding protein FBP54, fibronectin-binding protein FbaA, M protein type 1 Emml, M protein type 6 Emm6, immunoglobulin-binding protein 35 Sib35, Surface protein R28 Spr28, superoxide dismutase SOD, C5a peptidase ScpA, antigen I / II Agl / I I, adhesin AspA, G-related alpha2 -macroglobulin- binding protein GRAB, surface fibrillar protein M5 (Streptococcus pyogenes, Group A streptococcal infection); C protein P antigen, arginine deiminase proteins, adhesin Bib A, 105 kDAprotein BPS, surface antigens c, surface antigens R, surface antigens X, trypsin-resistant protein Rl, trypsin-resistant protein R3, trypsin-resistant protein R4, surface immunogenic protein Sip, surface protein Rib, Leucine-rich repeats protein LrrG, serine-rich repeat protein Srr-2, C protein alpha-antigen Bea, Beta antigen Bag, surface antigen Epsilon, alpha-like protein ALP1, alpha-like protein ALP5 surface antigen delta, alpha-like protein ALP2, alpha-like protein ALP3, alpha-like protein ALP4, Cbeta protein Bac (Streptococcus agalactiae, Group B streptococcal infection); transferrin-binding protein 2 Tbp2, phosphatase P4, outer membrane protein P6, peptidoglycan- associated lipoprotein Pal, protein D, protein E, adherence and penetration protein Hap, outer membrane protein 26 Omp26, outer membrane protein P5 (Fimbrin), outer membrane protein DI 5, outer membrane protein 0mpP2, 5 '-nucleotidase NucA, outer membrane protein Pl, outer membrane protein P2, outer membrane lipoprotein Pep, Lipoprotein E, outer membrane protein P4, fuculokinase FucK, [Cu,Zn]-superoxide dismutase SodC, protease HtrA, protein 0145, alphagalactosylceramide (Haemophilus influenzae, Haemophilus influenzae infection); polymerase 3D, viral capsid protein VP1, viral capsid protein VP2, viral capsid protein VP3, viral capsid protein VP4, protease 2 A, protease 3C (Enteroviruses, mainly Coxsackie A virus and Enterovirus 71 (EV71), Hand, foot and mouth disease (HFMD)); RNA polymerase L, protein L, glycoprotein Gn, glycoprotein Gc, nucleocapsid protein 5, envelope glycoprotein Gl, nucleoprotein NP, protein N, polyprotein M (Sin Nombre virus, Hantavirus, Hantavirus Pulmonary Syndrome (HPS)); heat shock protein HspA, heat shock protein HspB, citrate synthase GItA, protein UreB, heat shock protein Hsp60, neutrophil-activating protein NAP, catalase KatA, vacuolating cytotoxin VacA, urease alpha UreA, urease beta Ureb, protein CpnlO, protein groES, heat shock protein HsplO, protein MopB, cytotoxicity-associated 10 kDa protein CAG, 36 kDa antigen, beta-lactamase HcpA, Beta-lactamase HcpB (Helicobacter pylori, Helicobacter pylori infection); integral membrane proteins, aggregation-prone proteins, O-antigen, toxin-antigens Stx2B, toxin-antigen StxlB, adhesion-antigen fragment Int28, protein EspA, protein EspB, Intimin, protein Tir, protein IntC300, protein Eae (Escherichia coli 0157 :H7 , 0111 and 0104:H4, Hemolytic-uremic syndrome (HUS)); RNA polymerase L, protein L, glycoprotein Gn, glycoprotein Gc, nucleocapsid protein 5, envelope glycoprotein Gl, nucleoprotein NP, protein N, polyprotein M (Bunyaviridae family, Hemorrhagic fever with renal syndrome (HFRS)); glycoprotein G, matrix protein M, nucleoprotein N, fusion protein F, polymerase L, protein W, protein C, phosphoprotein p, non- structural protein V (Henipavirus (Hendra virus Nipah virus), Henipavirus infections); polyprotein, glycoproten Gp2, hepatitis A surface antigen HBAg, protein 2A, virus protein VP1, virus protein VP2, virus protein VP3, virus protein VP4, protein P1B, protein P2A, protein P3AB, protein P3D (Hepatitis A Virus, Hepatitis A); hepatitis B surface antigen HBsAg, Hepatitis B core antigen HbcAg, polymerase, protein Hbx, preS2 middle surface protein, surface protein L, largeS protein, virus protein VP1, virus protein VP2, virus protein VP3, virus protein VP4 (Hepatitis B Virus (HBV), Hepatitis B); envelope glycoprotein El gp32 gp35 envelope glycoprotein E2 NS1 gp68 gp70, capsid protein C core protein Core, polyprotein, virus protein VP1, virus protein VP2, virus protein VP3, virus protein VP4, antigen G, protein NS3, protein NSSA, (Hepatitis C Virus, Hepatitis C); virus protein VP1, virus protein VP2, virus protein VP3, virus protein VP4, large hepatitis delta antigen, small hepatitis delta antigen (Hepatitis D Virus, Hepatitis D); virus protein VP1, virus protein VP2, virus protein VP3, virus protein VP4, capsid protein E2 (Hepatitis E Virus, Hepatitis E); glycoprotein L ULI, uracil-DNA glycosylase UL2, protein UL3, protein UL4, DNA replication protein UL5, portal protein UL6, virion maturation protein UL7, DNA helicase UL8, replication origin-binding protein UL9, glycoprotein M UL10, protein UL11, alkaline exonuclease ULI 2, serine-threonine protein kinase ULI 3, tegument protein ULI 4, terminase ULI 5, tegument protein ULI 6, protein ULI 7, capsid protein VP23 ULI 8, major capsid protein VP5 ULI 9, membrane protein UL20, tegument protein UL21, Glycoprotein H (UL22), Thymidine Kinase UL23, protein UL24, protein UL25, capsid protein P40 (UL26, VP24, VP22A), glycoprotein B (UL27), ICP18.5 protein (UL28), major DNA-binding protein ICP8 (UL29), DNA polymerase UL30, nuclear matrix protein UL31, envelope glycoprotein UL32, protein UL33, inner nuclear membrane protein UL34, capsid protein VP26 (UL35), large tegument protein UL36, capsid assembly protein UL37, VP19C protein (UL38), ribonucleotide reductase (Large subunit) UL39, ribonucleotide reductase (Small subunit) UL40, tegument protein / virion host shutoff VHS protein (UL41), DNA polymerase processivity factor UL42, membrane protein UL43, glycoprotein C (UL44), membrane protein UL45, tegument proteins VP11 / 12 (UL46), tegument protein VP 13 / 14 (UL47), virion maturation protein VP 16 (UL48, Alpha- TIF), envelope protein UL49, dUTP diphosphatase UL50, tegument protein UL51, DNA helicase / primase complex protein UL52, glycoprotein K (UL53), transcriptional regulation protein 1E63 (ICP27, UL54), protein UL55, protein UL56, viral replication protein ICP22 (1E68, US1), protein U52, serine / threonine-protein kinase U53, glycoprotein G (U54), glycoprotein J (U55), glycoprotein D (U56), glycoprotein I (U57), glycoprotein E (U58), tegument protein U59, capsid / tegument protein US10, Vmw21 protein (US11), ICP47 protein (IE12, US12), major transcriptional activator ICP4 (1E175, RSI), E3 ubiquitin ligase ICPO (IE110), latency-related protein 1 LRP1, latency-related protein 2 LRP2, neurovirulence factor RL1 (ICP34.5), latency-associated transcript LAT (Herpes simplex virus 1 and 2 (HSV-1 and HSV-2), Herpes simplex); heat shock protein Hsp60, cell surface protein H1C, dipeptidyl peptidase type IV DppIV, M antigen, 70 kDa protein, 17 kDa histone-like protein (Histoplasma capsulatum, Histoplasmosis); fatty acid and retinol binding protein-1 FAR-1, tissue inhibitor of metalloproteinase TIMP (TMP), cysteine proteinase ACEY-1, cysteine proteinase ACCP-1, surface antigen Ac- 16, secreted protein 2 ASP-2, metalloprotease 1 MTP-1, aspartyl protease inhibitor API-1, surface-associated antigen SAA- 1, surface-associated antigen SAA-2, adult-specific secreted factor Xa, serine protease inhibitor anticoagulant AP, cathepsin D-like aspartic protease ARR-1, 5-transferase GST, aspartic protease APR-1, acetylcholinesterase AChE (Ancylostoma duodena le and Vecator americanus. Hookworm infection); protein NS 1, protein NP1, protein VP1, protein VP2, protein VP3 (Human bocavirus (HBoV), Human bocavirus infection); major surface protein 2 MSP2, major surface protein 4 MSP4, MSP variant SGV1, MSP variant SGV2, outer membrane protein OMP, outer membrane protein 19 OMP- 19, major antigenic protein MAPI, major antigenic protein MAP 1-2, major antigenic protein MAP1B, major antigenic protein MAPI-3, Erum2510 coding protein, protein GroEL, protein GroES, 30-kDA major outer membrane proteins, GE 100-kDa protein, GE 130-kDa protein, GE 160-kDa protein (Ehrlichia ewingii, Human ewingii ehrlichiosis); major surface proteins 1-5 (MSPla, MSPlb, MSP2, MSP3, MSP4, MSP5), type IV secretion system proteins VirB2, VirB7, VirBl l, VirD4 (Anaplasma phagocy tophilum, Human granulocytic anaplasmosis (HGA)); protein NS1, small hydrophobic protein N52, SH protein, fusion protein F, glycoprotein G, matrix protein M, matrix protein M2-1, matrix protein M2-2, phosphoprotein P, nucleoprotein N, polymerase L (Human metapneumovirus (hMPV), Human metapneumovirus infection); major surface protein 2 MSP2, major surface protein 4 MSP4, MSP variant SGV1, MSP variant SGV2, outer membrane protein OMP, outer membrane protein 19 OMP- 19, major antigenic protein MAPI, major antigenic protein MAPI-2, major antigenic protein MAP1B, major antigenic protein MAP 1-3, Erum2510 coding protein, protein GroEL, protein GroES, 30-kDA major outer membrane proteins, GE 100-kDa protein, GE 130-kDa protein, GE 160-kDa protein (Ehrlichia chaffeensis, Human monocytic ehrlichiosis); replication protein EL, regulatory protein E2, protein E3, protein E4, protein ES, protein E6, protein E7, protein E8, major capsid protein LI, minor capsid protein L2 (Human papillomavirus (HPV), Human papillomavirus (HPV) infection); fusion protein F, hemagglutinin-neuramidase HN, glycoprotein G, matrix protein M, phosphoprotein P, nucleoprotein N, polymerase L (Human parainfluenza viruses (HPIV), Human parainfluenza virus infection); Hemagglutinin (HA), Neuraminidase (NA), Nucleoprotein (NP), Ml protein, M2 protein, NS1 protein, NS2 protein (NEP protein: nuclear export protein), PA protein, PB1 protein (polymerase basic 1 protein), PB1-F2 protein and PB2 protein (Orthomyxoviridae family, Influenza virus (flu)); genome polyprotein, protein E, protein M, capsid protein C (Japanese encephalitis virus, Japanese encephalitis); RTX toxin, type IV pili, major pilus subunit PilA, regulatory transcription factors PilS and PilR, protein sigma54, outer membrane proteins (Kingella kingae, Kingella kingae infection); prion protein (Kuru prion, Kuru); nucleoprotein N, polymerase L, matrix protein Z, glycoprotein GP (Lassa virus, Lassa fever); peptidoglycan-associated lipoprotein PAL, 60 kDa chaperonin Cpn60 (groEL, HspB), typeIV pilin PilE, outer membrane protein MIP, major outer membrane protein MompS, zinc metalloproteinase MSP (Legionella pneumophila, Legionellosis (Legionnaires' disease, Pontiac fever)); P4 nuclease, protein WD, ribonucleotide reductase M2, surface membrane glycoprotein Pg46, cysteine proteinase CP, glucose-regulated protein 78 GRP-78, stage-specific S antigen-like protein A2, ATPase Fl, beta-tubulin, heat shock protein 70 Hsp70, KMP-11, glycoprotein GP63, protein BT1, nucleoside hydrolase NH, cell surface protein Bl, ribosomal protein Pl -like protein Pl, sterol 24-c-methyltransferase SMT, LACK protein, histone Hl, SPB1 protein, thiol specific antioxidant TSA, protein antigen STH, signal peptidase SP, histone H2B, surface antigen PSA-2, cystein proteinase b Cpb (Leishmania genus, Leishmaniasis); major membrane protein I, serine- rich antigen-45 kDa, 10 kDa caperonin GroES, HSP kDa antigen, amino-oxononanoate synthase AONS, protein recombinase A RecA, AcetyL / propionyl-coenzyme A carboxylase alpha, alanine racemase, 60 kDa chaperonin 2, ESAT-6-like protein EcxB (L-ESAT-6), protein Lsr2, protein ML0276, Heparin-binding hemagglutinin HBHA, heat-shock protein 65 Hsp65, mycPl or ML0041 coding protein htrA2 or ML0176 coding protein htrA4 or ML2659 coding protein, gcp or ML0379 coding protein, clpC or ML0235 coding protein (Mycobacterium leprae and Mycobacterium lepromatosis, Leprosy); outer membrane protein LipL32, membrane protein LIC10258, membrane protein LP30, membrane protein LIC12238, Ompa-like protein Lsa66, surface protein LigA, surface protein LigB, major outer membrane protein OmpLl, outer membrane protein LipL41, protein LigAni, surface protein LcpA, adhesion protein LipL53, outer membrane protein UpL32, surface protein Lsa63, flagellin FlaBl, membrane lipoprotein LipL21, membrane protein pL40, leptospiral surface adhesin Lsa27, outer membrane protein OmpL36, outer membrane protein OmpL37, outer membrane protein OmpL47, outer membrane protein OmpL54, acyltransferase LpxA (Leptospira genus, Leptospirosis); listeriolysin O precursor Hly (LLO), invasion-associated protein lap (P60), Listeriolysin regulatory protein PrfA, Zinc metalloproteinase Mpl, Phosphatidylinositol-specific phospholipase C PLC (PIcA, PlcB), 0- acetyltransferase Oat, ABC-transporter permease Im.G_1771, adhesion protein LAP, LAP receptor Hsp60, adhesin LapB, haemolysin listeriolysin O LLO, protein ActA, Intemalin A InIA, protein InIB (Listeria monocytogenes, Listeriosis); outer surface protein A OspA, outer surface protein OspB, outer surface protein OspC, decorin binding protein A DbpA, decorin binding protein B DbpB, flagellar filament 41 kDa core protein Fla, basic membrane protein A Bmp A (Immunodominant antigen P39), outer surface 22 kDa lipoprotein precursor (antigen IPLA7), variable surface lipoprotein vlsE (usually Borrelia burgdorferi and other Borrelia species, Lyme disease (Lyme borreliosis)); venom allergen homolog-like protein VAL-1, abundant larval transcript ALT-1, abundant larval transcript ALT-2, thioredoxin peroxidase TPX, vespid allergen homologue VAH, thi ordoxin peroxidase 2 TPX-2, antigenic protein SXP (peptides N, Nl, N2,and N3), activation associated protein-1 ASP-1, thioredoxin TRX, transglutaminase BmTGA, glutathione-S-transferases GST, myosin, vespid allergen homologue VAH, 175 kDa collagenase, glyceraldehyde-3 -phosphate dehydrogenase GAPDH, cuticular collagen Col -4, Secreted Larval Acidic Proteins SLAPs, chitinase CHI-1, maltose binding protein MBP, glycolytic enzyme fructose- 1,6-bisphosphate aldolase Fba, tropomyosin TMY-1, nematode specific gene product OvB20, onchocystatin CPI-2, protein Cox-2 (Wuchereria bancrofti and Brugia malctyi. Lymphatic filariasis (Elephantiasis)); glycoprotein GP, matrix protein polymerase L, nucleoprotein N (Lymphocytic choriomeningitis virus (LCMV), Lymphocytic choriomeningitis); thrombospondin-related anonymous protein TRAP, SSP2 Sporozoite surface protein 2, apical membrane antigen 1 AMA1, rhoptry membrane antigen RMA1, acidic basic repeat antigen ABRA, cell-traversal protein PF, protein Pvs25, merozoite surface protein 1 MSP-1, merozoite surface protein 2 MSP-2, ring-infected erythrocyte surface antigen RESALiver stage antigen 3 LSA-3, protein Eba-175, serine repeat antigen 5 SERA-5, circumsporozoite protein CS, merozoite surface protein 3 MSP3, merozoite surface protein 8 MSP5, enolase PF10, hepatocyte erythrocyte protein 17 kDa HEP 17, erythrocyte membrane protein 1 EMP1, protein Kbetamerozoite surface protein 4 / 5 MSP 4 / 5, heat shock protein Hsp90, glutamate-rich protein GLURP, merozoite surface protein 4 MSP-4, protein STARP, circumsporozoite protein-related antigen precursor CRA (Plasmodium genus, Malaria); nucleoprotein N, membrane-associated protein VP24, minor nucleoprotein VP30, polymerase cofactor VP35, polymerase L, matrix protein VP40, envelope glycoprotein GP (Marburg virus, Marburg hemorrhagic fever (MHF)); protein C, matrix protein M, phosphoprotein P, non-structural protein V, hemagglutinin glycoprotein H, polymerase L, nucleoprotein N, fusion protein F (Measles virus, Measles); members of the ABC transporter family (LoIC, OppA, and PotF), putative lipoprotein releasing system transmembrane protein LoIC / E, flagellin FliC, Burkholderia intracellular motility A BimA, bacterial Elongation factor- Tu EF-Tu, 17 kDa OmpA-like protein, boaA coding protein, boaB coding protein (Burkholderia pseudomallei, Melioidosis (Whitmore's disease)); pilin proteins, minor pilin-associated subunit pilC, major pilin subunit and variants pilE, pilS, phase variation protein porA, Porin B PorB, protein TraD, Neisserial outer membrane antigen H.8, 70 kDa antigen, major outer membrane protein PI, outer membrane proteins PIA and PIB, W antigen, surface protein A NspA, transferrin binding protein TbpA, transferrin binding protein TbpB PBP2, mtrR coding protein, ponA coding protein, membrane permease FbpBC, FbpABC protein system, LbpAB proteins, outer membrane protein Opa, outer membrane transporter FetA, iron-repressed regulator MpeR, factor H-binding protein fHbp, adhesin NadA, protein NhbA, repressor FarR (Neisseria meningitidis, Meningococcal disease); 66 kDa protein, 22 kDa protein (usually Metagonimus yokagawai, Metagonimiasis); polar tube proteins (34, 75, and 170 kDa in Glugea, 35, 55 and 150 kDa inEncephalitozoon), kinesin-related protein, RNA polymerase II largest subunit, similar of integral membrane protein YIP A, anti-silencing protein 1, heat shock transcription factor HSF, protein kinase, thymidine kinase, NOP-2 like nucleolar protein (Microsporidia phylum, Microsporidiosis); CASP8 and FADD-like apoptosis regulator, Glutathione peroxidase GPX1, RNA helicase NPH-II NPH2, Poly(A) polymerase catalytic subunit PAPL, Major envelope protein P43K, early transcription factor 70 kDa subunit VETFS, early transcription factor 82 kDa subunit VETFL, metalloendopeptidase Gl-type, nucleoside triphosphatase I NPH1, replication protein A28-like MC134L, RNA polymerase 7 kDa subunit RPO7 (Molluscum contagiosum virus (MCV), Molluscum contagiosum (MC)); matrix protein M, phosphoprotein P / V, small hydrophobic protein SH, nucleoprotein N, protein V, fusion glycoprotein hemagglutininneuraminidase HN, RNA polymerase L (Mumps virus, Mumps); Outer membrane proteins OM, cell surface antigen OmpA, cell surface antigen OmpB (sca5), cell surface protein SCA4, cell surface protein SCA1, intracytoplasmic protein D, crystalline surface layer protein SLP, protective surface protein antigen SPA (Rickettsia typhi, Murine typhus (Endemic typhus)); adhesin Pl, adhesion P30, protein pl 16, protein P40, cytoskeletal protein HMW1, cytoskeletal protein HMW2, cytoskeletal protein HMW3, MPN 152 coding protein, MPN426 coding protein, MPN456 coding protein, MPN-500coding protein (Mycoplasma pneumoniae, Mycoplasma pneumonia); NocA, Iron dependent regulatory protein, VapA, VapD, VapF, VapG, caseinolytic protease, filament tip-associated 43-kDa protein, protein P24, protein P61, 15-kDa protein, 56-kDa protein (usually Nocardia asteroides and other Nocardia species, Nocardiosis); venom allergen homolog-like protein VAL-1, abundant larval transcript ALT-1, abundant larval transcript ALT- 2, thioredoxin peroxidase TPX, vespid allergen homologue VAH, thiordoxin peroxidase 2 TPX- 2, antigenic protein SXP (peptides N, Nl, N2, and N3), activation associated protein-1 ASP-1, Thioredoxin TRX, transglutaminase BmTGA, glutathione-S-transferases GST, myosin, vespid allergen homologue VAH, 175 kDa collagenase, glyceraldehyde-3 -phosphate dehydrogenase GAPDH, cuticular collagen Col-4, Secreted Larval Acidic Proteins SLAPs, chitinase CHI-1, maltose binding protein MBP, glycolytic enzyme fructose- 1,6-bisphosphate aldolase Fba, tropomyosin TMY-1, nematode specific gene product OvB20, onchocy statin CPL2, Cox-2 (Onchocerca volvulus, Onchocerciasis (River blindness)); 43 kDa secreted glycoprotein, glycoprotein gpO, glycoprotein gp75, antigen Pb27, antigen Pb40, heat shock protein Hsp65, heat shock protein Hsp70, heat shock protein Hsp90, protein PIO, triosephosphate isomerase TPI, N- acetyl-glucosamine-binding lectin Paracoccin, 28 kDa protein Pb28 (Paracoccidioides brasiliensis, Paracoccidioidomycosis (South American blastomycosis)); 28-kDa cruzipain-like cystein protease Pw28CCP (usually Paragonimus westermani and other Paragonimus species, Paragonimiasis); outer membrane protein OmpH, outer membrane protein Omp28, proteinPM1539, protein PM0355, protein PM1417, repair protein MutL, protein BcbC, protein PM0305, formate dehydrogenase-N, protein PM0698, protein PM1422, DNA gyrase, lipoprotein PIpE, adhesive protein Cp39, heme acquisition system receptor HasR, 39 kDa capsular protein, iron- regulated OMP IROMP, outer membrane protein OmpA87, fimbrial protein Ptf, fimbrial subunit protein PtfA, transferrin binding protein Tbpl, esterase enzyme MesA, Pasteurella multocida toxin PMT, adhesive protein Cp39 (Pasteurella genus, Pasteurellosis); “filamentous hemagglutinin FhaB, adenylate cyclase CyaA, pertussis toxin subunit 4 precursor PtxD, pertactin precursor Prn, toxin subunit 1 PtxA, protein Cpn60, protein brkA, pertussis toxin subunit 2 precursor PtxB, pertussis toxin subunit 3 precursor PtxC, pertussis toxin subunit 5 precursor PtxE, pertactin Pm, protein Fim2, protein Fim3;” (Bordetella pertussis, Pertussis (Whooping cough)); “Fl capsule antigen, virulence-associated V antigen, secreted effector protein LcrV, V antigen, outer membrane protease Pla, secreted effector protein YopD, putative secreted protein-tyrosine phosphatase YopH, needle complex major subunit YscF, protein kinase YopO, putative autotransporter protein YapF, inner membrane ABC-transporter YbtQ (Irp7), putative sugar binding protein YP00612, heat shock protein 90 HtpG, putative sulfatase protein YdeN, outermembrane lipoprotein carrier protein LoIA, secretion chaperone Yer A, putative lipoprotein YP00420, hemolysin activator protein HpmB, pesticin / yersiniabactin outer membrane receptor Psn, secreted effector protein YopE, secreted effector protein YopF, secreted effector protein YopK, outer membrane protein YopN outer membrane protein YopM, Coagulase / fibrinolysin precursor Pla;” (Yersinia pestis. Plague); protein PhpA, surface adhesin PsaA, pneumolysin Ply, ATP-dependent protease CIp, lipoate-protein ligase LpIA, cell wall surface anchored protein psrP, sortase SrtA, glutamyl-tRNA synthetase GItX, choline binding protein A CbpA, pneumococcal surface protein A PspA, pneumococcal surface protein C PspC, 6-phosphogluconate dehydrogenase Gnd, iron-binding protein PiaA, Murein hydrolase LytB, proteon LytC, protease Al (Streptococcus pneumoniae, Pneumococcal infection); major surface protein B, kexin-like protease KEX1, protein A12, 55 kDa antigen P55, major surface glycoprotein Msg (Pneumocystis jirovecii, Pneumocystis pneumonia (PCP)); genome polyprotein, polymerase 3D, viral capsid protein VP1, viral capsid protein VP2, viral capsid protein VP3, viral capsid protein VP4, protease 2A, protease 3C (Poliovirus, Poliomyelitis); protein Nfal, exendin-3, secretory lipase, cathepsin B-like protease, cysteine protease, cathepsin, peroxiredoxin, protein CrylAc (usually Naegleria fowleri, Primary amoebic meningoencephalitis (PAM)); agnoprotein, large T antigen, small T antigen, major capsid protein VP1, minor capsid protein Vp2 (JC virus, Progressive multifocal leukoencephalopathy); low calcium response protein E LCrE, chlamydial outer protein N CopN, serine / threonine-protein kinase PknD, acyl-carrier-protein S-malonyltransferase FabD, singlestranded DNA-binding protein Ssb, major outer membrane protein MOMP, outer membraneprotein 2 0mp2, polymorphic membrane protein family (Pmpl, Pmp2, Pmp3, Pmp4, Pmp5, Pmp6, Pmp7, Pmp8, Pmp9, PmplO, Pmpl l, Pmpl2, Pmpl3, Pmpl4, Pmpl5, Pmpl6, Pmpl7, Pmpl 8, Pmpl 9, Pmp20, Pmp21) (Chlamydophila psittaci, Psittacosis); outer membrane protein Pl, heat shock protein B HspB, peptide ABC transporter, GTP -binding protein, protein IcmB, ribonuclease R, phosphatas SixA, protein DsbD, outer membrane protein ToIC, DNA-binding protein PhoB, ATPase DotB, heat shock protein B HspB, membrane protein Coml, 28 kDa protein, DNA-3 -methyladenine glycosidase I, pouter membrane protein OmpH, outer membrane protein AdaA, glycine cleavage system T-protein (Coxiella burnetii, Q fever); nucleoprotein N, large structural protein L, phophoprotein P, matrix protein M, glycoprotein G (Rabies virus, Rabies); fusionprotein F, nucleoprotein N, matrix protein M, matrix protein M2-1, matrix protein M2-2, phophoprotein P, small hydrophobic protein SH, major surface glycoprotein G, polymerase L, non-structural protein 1 NS1, non- structural protein 2 NS2 (Respiratory syncytial virus (RSV), Respiratory syncytial virus infection); genome polyprotein, polymerase 3D, viral capsid protein VP1, viral capsid protein VP2, viral capsid protein VP3, viral capsid protein VP4, protease 2 A, protease 3C (Rhinovirus, Rhinovirus infection); outer membrane proteins OM, cell surface antigen OmpA, cell surface antigen OmpB (sca5), cell surface protein SCA4, cell surface protein SCA1, protein PS120, intracytoplasmic protein D, protective surface protein antigen SPA (Rickettsia genus, Rickettsial infection); outer membrane proteins OM, cell surface antigen OmpA, cell surface antigen OmpB (sca5), cell surface protein SCA4, cell surface protein SCA1, intracytoplasmic protein D (Rickettsia akari. Rickettsialpox); envelope glycoprotein GP, polymerase L, nucleoprotein N, non-structural protein NSS (Rift Valley fever virus, Rift Valley fever (RVF)); outer membrane proteins OM, cell surface antigen OmpA, cell surface antigen OmpB (sca5), cell surface protein SCA4, cell surface protein SC Al, intracytoplasmic protein D (Rickettsia rickeUsii, Rocky mountain spotted fever (RMSF)); non-structural protein 6 N56, non- structural protein 2 N52, intermediate capsid protein VP6, inner capsid protein VP2, non- structural protein 3 NS3, RNA-directed RNA polymerase L, protein VP3, non-structural protein 1 NS1, non-structural protein 5 N55, outer capsid glycoprotein VP7, non-structural glycoprotein 4 N54, outer capsid protein VP4; (Rotavirus, Rotavirus infection); polyprotein P200, glycoprotein El, glycoprotein E2, protein N52, capsid protein C (Rubella virus, Rubella); chaperonin GroEL (Mop A), inositol phosphate phosphatase SopB, heat shock protein HslU, chaperone protein DnaJ, protein TviB, protein IroN, flagellin FliC, invasion protein SipC, glycoprotein gp43, outer membrane protein LamB, outer membrane protein PagC, outer membrane protein ToIC, outer membrane protein NmpC, outer membrane protein FadL, transport protein SadA, transferase WgaP, effector proteins SifA, SteC, SseL, SseJ and SseF (Salmonella genus, Salmonellosis)' ,' protein 14, non-structural protein NS7b, non-structural protein NS8a, protein 9b, protein 3a,nucleoprotein N, non-structural protein NS3b, non- structural protein N56, protein 7a, non- structural protein NS8b, membrane protein M, envelope small membrane protein EsM, replicase polyprotein la, spike glycoprotein S, replicase polyprotein lab; SARS coronavirus, SARS (Severe Acute Respiratory Syndrome)); serin protease, Atypical Sarcoptes Antigen 1 ASAI, glutathione 5 -transferases GST, cystein protease, serine protease, apolipoprotein (Sarcoptes scabiei, Scabies); glutathione 5 -transferases GST, paramyosin, hemoglbinase SM32, major egg antigen, 14 kDa fatty acid-binding protein Sml4, major larval surface antigen P37, 22.6 kDa tegumental antigen, calpain CANP, triphospate isomerase Tim, surface protein 9B, outer capsid protein VP2, 23 kDa integral membrane protein Sm23, Cu / Zn-superoxide dismutase, glycoprotein Gp, myosin (Schistosoma genus, Schistosomiasis (Bilharziosis)); 60 kDa chaperonin, 56 kDa type-specific antigen, pyruvate phosphate dikinase, 4-hydroxybenzoate octaprenyltransferase (Orientia tsutsugamushi, Scrub typhus); dehydrogenase GuaB, invasion protein Spa32, invasin IpaA, invasin IpaB, invasin IpaC, invasin IpaD, invasin IpaH, invasin IpaJ (Shigella genus, Shigellosis (Bacillary dysentery)); protein P53, virion protein US10 homolog, transcriptional regulator 1E63, transcriptional transactivator 1E62, protease P33, alpha trans-inducing factor 74 kDa protein, deoxyuridine 5 '-triphosphate nucleotidohydrolase, transcriptional transactivator 1E4, membrane protein UL43 homolog, nuclear phosphoprotein UL3 homolog, nuclear protein UL4 homolog, replication origin-binding protein, membrane protein 2, phosphoprotein 32, protein 57, DNA polymerase processivity factor, portal protein 54, DNA primase, tegument protein ULI 4 homolog, tegument protein UL21 homolog, tegument protein UL55 homolog, tripartite terminase subunit UL33 homolog, tripartite terminase subunit ULI 5 homolog, capsid-binding protein 44, virionpackaging protein 43 (Varicella zoster virus (VZV), Shingles (Herpes zoster)); truncated 3-beta hydroxy-5-ene steroid dehydrogenase homolog, virion membrane protein A13, protein A19, protein A31, truncated protein A35 homolog, protein A37.5 homolog, protein A47, protein A49, protein A51, semaphorin-like protein A43, serine proteinase inhibitor 1, serine proteinase inhibitor 2, serine proteinase inhibitor 3, protein A6, protein B15, protein Cl, protein C5, protein C6, protein F7, protein F8, protein F9, protein Fl 1, protein F14, protein F15, protein F16 (Variola major or Variola minor, Smallpox (Variola)); adhesin / glycoprotein gp70, proteases (Sporothrix schenckir Sporotrichosis); heme-iron binding protein IsdB, collagen adhesin Cna, clumping factor A ClfA, protein MecA, fibronectin-binding protein A FnbA, enterotoxin type A EntA, enterotoxin type B EntB, enterotoxin type C EntCl, enterotoxin type C EntC2, enterotoxin type D EntD, enterotoxin type E EntE, Toxic shock syndrome toxin-1 TSST-1, Staphylokinase, Penicillin binding protein 2a PBP2a (MecA), secretory antigen SssA (Staphylococcus genus, Staphylococcal food poisoning); heme-iron binding protein IsdB, collagen adhesin Cna, clumping factor A ClfA, protein MecA, fibronectin-binding protein A FnbA, enterotoxin type A EntA,enterotoxin type B EntB, enterotoxin type C EntCl, enterotoxin type C EntC2, enterotoxin type D EntD, enterotoxin type E EntE, Toxic shock syndrome toxin-1 TSST-1, Staphylokinase, Penicillin binding protein 2a PBP2a (MecA), secretory antigen SssA (Staphylococcus genus e.g. aureus, Staphylococcal infection); antigen Ss-IR, antigen NIE, strongylastacin, Na+-K+ ATPase Sseat-6, tropomysin SsTmy-1, protein LEC-5, 41 kDa aantigen P5, 41-kDa larval protein, 31-kDa larval protein, 28-kDa larval protein (Strongyloides slercoralis. Strongyloidiasis); glycerophosphodiester phosphodiesterase GlpQ (Gpd), outer membrane protein TmpB, protein Tp92, antigen TpFl, repeat protein Tpr, repeat protein F TprF, repeat protein G TprG, repeat protein I Tprl, repeat protein J TprJ, repeat protein KTprK, treponemal membrane protein A TmpA, lipoprotein, 15 kDa Tppl5, 47 kDa membrane antigen, miniferritin TpFl, adhesin Tp0751, lipoprotein TP0136, protein TpN17, protein TpN47, outer membrane protein TP0136, outer membrane protein TP0155, outer membrane protein TP0326, outer membrane protein TP0483, outer membrane protein TP0956 (Treponema pallidum, Syphilis); Cathepsin L-like proteases, 53 / 25-kDa antigen, 8 kDa family members, cysticercus protein with a marginal trypsin-like activity TsAg5, oncosphere protein TSOL18, oncosphere protein TSOL45-1A, lactate dehydrogenase A LDHA, lactate dehydrogenase B LDHB Taenia genus, Taeniasis); tetanus toxin TetX, tetanus toxin C TTC, 140 kDa S layer protein, flavoprotein beta-subunit CT3, phospholipase (lecithinase), phosphocarrier protein HPr (Clostridium tetani, Tetanus (Lockjaw)); genome polyprotein, protein E, protein M, capsid protein C (Tick-borne encephalitis virus (TBEV), Tick-borne encephalitis); 58-kDa antigen, 68-kDa antigens, Toxocara larvae excretory- secretory antigen TES, 32-kDa glycoprotein, glycoprotein TES-70, glycoprotein GP31, excretory- secretory antigen TcES-57, perienteric fluid antigen Pe, soluble extract antigens Ex, excretory / secretory larval antigens ES, antigen TES- 120, polyprotein allergen TBA-1, cathepsin L-like cysteine protease c-cpl-1, 26-kDa protein (Toxocara canis or Toxocara cati, Toxocariasis (Ocular Larva Migrans (OLM) and Visceral Larva Migrans (VLM))); microneme proteins (MIC1, MIC2, MIC3, MIC4, MIC5, MICE, MIC7, MICE), rhoptry protein Rop2, rhoptry proteins (Ropl, Rop2, Rop3, Rop4, Rop5, Rop6, Rop7, Ropl6, Rjopl7), protein SRI, surface antigen P22, major antigen p24, major surface antigen p30, dense granule proteins (GRA1, GRA2, GRA3, GRA4, GRA5, GRA6, GRA7, GRAB, GRA9, GRA10), 28 kDa antigen, surface antigen SAG1, SAG2 related antigen, nucleoside-triphosphatase 1, nucleoside-triphosphatase 2, protein Stt3, HesB-like domain-containing protein, rhomboid-like protease 5, toxomepsin 1 (Toxoplasma gondii, Toxoplasmosis); 43 kDa secreted glycoprotein, 53 kDa secreted glycoprotein, paramyosin, antigen Ts21, antigen Ts87, antigen p46000, TSL-1 antigens, caveolin-1 CAV-1, 49 kDa newborn larva antigen, prosaposin homologue, serine protease, serine proteinase inhibitor, 45-kDa glycoprotein Gp45 (Trichinella spiralis, Trichinellosis); Myb-like transcriptional factors (Mybl,Myb2, Myb3), adhesion protein AP23, adhesion protein AP33, adhesin protein AP33-3, adhesins AP51, adhesin AP65, adhesion protein AP65-1, alpha-actinin, kinesin-associated protein, teneurin, 62 kDa proteinase, subtili sin-like serine protease SUB1, cysteine proteinase gene 3 CP3, alpha-enolase Enol, cysteine proteinase CP30, heat shock proteins (Hsp70, Hsp60) immunogenic protein P270, (Trichomonas vaginalis, Trichomoniasis); beta-tubulin, 47-kDa protein, secretory leucocyte-like proteinase- 1 SLP-1, 50-kDa protein TT50, 17 kDa antigen, 43 / 47 kDa protein (Trichuris Irichiura, Trichuriasis (Whipworm infection)); protein ESAT-6 (EsxA), 10 kDa filtrate antigen EsxB, secreted antigen 85-B FBPB, fibronectin-binding protein A FbpA (Ag85A), serine protease PepA, PPE family protein PPE18, fibronectin-binding protein D FbpD, immunogenic protein MPT64, secreted protein MPT51, catalase-peroxidase-peroxynitritase T KATG, periplasmic phosphate-binding lipoprotein PSTS3 (PBP-3, Phos-1), iron-regulated heparin binding hemagglutinin Hbha, PPE family protein PPE14, PPE family protein PPE68, protein Mtb72F, protein Apa, immunogenic protein MPT63, periplasmic phosphate-binding lipoprotein PSTS1 (PBP-1), molecular chaperone DnaK, cell surface lipoprotein Mpt83, lipoprotein P23, phosphate transport system permease protein pstA, 14 kDa antigen, fibronectin-binding protein C FbpCl, Alanine dehydrogenase TB43, Glutamine synthetase 1, ESX-1 protein, protein CFP10, TB10.4 protein, protein MPT83, protein MTB12, protein MTBE, Rpf-like proteins, protein MTB32, protein MTB39, crystallin, heat-shock protein HSP65, protein PST- S(usually Mycobacterium tuberculosis, Tuberculosis); outer membrane protein Fob A, outer membrane protein FobB, intracellular growth locus IgICl, intracellular growth locus IgIC2, aminotransferase Wbtl, chaperonin GroEL, 17 kDa major membrane protein TUL4, lipoprotein LpnA, chitinase family 18 protein, isocitrate dehydrogenase, Nif3 family protein, type IV pili glycosylation protein, outer membrane protein toIC, FAD binding family protein, type IV pilin multimeric outer membrane protein, two component sensor protein KdpD, chaperone protein DnaK, protein TolQ (Francisella tularensis, Tularemia); “MB antigen, urease, protein GyrA, protein GyrB, protein ParC, protein ParE, lipid associated membrane proteins LAMP, thymidine kinase TK, phospholipase PL-A1, phospholipase PL-A2, phospholipase PL-C, surface-expressed 96-kDa antigen;” (Ureaplasma urealyticum, Ureaplasma urealyticum infection); non- structural polyprotein, structural polyprotein, capsid protein CP, protein El, protein E2, protein E3, protease Pb, protease P2, protease P3 (Venezuelan equine encephalitis virus, Venezuelan equine encephalitis); glycoprotein GP, matrix protein Z, polymerase L, nucleoprotein N (Guanarito virus, Venezuelan hemorrhagic fever); polyprotein, protein E, protein M, capsid protein C, protease NS3, protein NS1, protein NS2A, protein AS2B, brotein NS4A, protein NS4B, protein NS5 (West Nile virus, West Nile Fever); cpasid protein CP, protein El, protein E2, protein E3, protease P2 (Western equine encephalitis virus, Western equine encephalitis); genome polyprotein, protein E,protein M, capsid protein C, protease NS3, protein NS1, protein NS2A, protein AS2B, protein NS4A, protein NS4B, protein NS5 (Yellow fever virus, Yellow fever); putative Yop targeting protein YobB, effector protein YopD, effector protein YopE, protein YopH, effector protein YopJ, protein translocation protein YopK, effector protein YopT, protein YpkA, flagellar biosynthesis protein FlhA, peptidase M48, potassium efflux system KefA, transcriptional regulator RovA, adhesin Ifp, translocator protein LcrV, protein PcrV, invasin Inv, outer membrane protein OmpF- like porin, adhesin YadA, protein kinase C, phospholipase Cl, protein PsaA, mannosy Itransferase- like protein WbyK, protein YscU, antigen YPMa (Yersinia pseudotuberculosis, Yersinia pseudotuberculosis infection); and effector protein YopB, 60 kDa chaperonin, protein WbcP, tyrosine-protein phosphatase YopH, protein YopQ, enterotoxin, Galactoside permease, reductase NrdE, protein YasN, Invasin Inv, adhesin YadA, outer membrane porin F OmpF, protein UspAl, protein EibA, protein Hia, cell surface protein Ail, chaperone SycD, protein LcrD, protein LcrG, protein LcrV, protein SycE, protein YopE, regulator protein Tye A, protein YopM, protein YopN, protein YopO, protein YopT, protein YopD, protease CIpP, protein MyfA, protein FilA, and protein PsaA (Yersinia enterocolitica, Yersiniosis).

[0192] The infectious agent can be a bacterium, a fungus, a virus, or a protist. The infectious agent can be a coronavirus (CoV) (e.g., an alphacoronavirus, a betacoronavirus, a gammacoronavirus, or a deltacoronavirus). The infectious agent can be selected from the group comprising Acinetobacter baumannii, Anaplasma genus, Anaplasma phagocytophilum, Ancylostoma braziliense, Ancylostoma duodenale, Arcanobacterium haemolyticum, Ascaris lumbricoides, Aspergillus genus, Astroviridae, Babesia genus, Bacillus anthracis, Bacillus cereus, Bartonella henselae, BK virus, Blastocystis hominis, Blastomyces dermatitidis, Bordetella pertussis, Borrelia burgdorferi, Borrelia genus, Borrelia spp, Brucella genus, Brugia malayi, Bunyaviridae family, Burkholderia cepacia and other Burkholderia species, Burkholderia mallei, Burkholderia pseudomallei, Caliciviridae family, Campylobacter genus, Candida albicans, Candida spp, Chlamydia trachomatis, Chlamydophila pneumoniae, Chlamydophila psittaci, CJD prion, Clonorchis sinensis, Clostridium botulinum, Clostridium difficile, Clostridium perfr ingens, Clostridium perfringens, Clostridium spp, Clostridium tetani, Coccidioides spp, coronaviruses, Corynebacterium diphtheriae, Coxiella burnetii, Crimean-Congo hemorrhagic fever virus, Cryptococcus neoformans, Cryptosporidium genus, Cytomegalovirus (CMV), Dengue viruses (DEN-1, DEN-2, DEN-3 and DEN-4), Dientamoeba fragilis, Ebolavirus (EBOV), Echinococcus genus, Ehrlichia chaffeensis, Ehrlichia ewingii, Ehrlichia genus, Entamoeba histolytica, Enterococcus genus, Enterovirus genus, Enteroviruses, mainly Coxsackie A virus and Enterovirus 71 (EV71), Epidermophyton spp, Epstein-Barr Virus (EBV), Escherichia coli 0157 :H7 , 0111 and 0104 :H4, Fasciola hepatica and Fasciola gigantica, FFI prion,Filarioidea superfamily, Filoviruses, Flaviviruses, Francisella tularensis, Fusobacterium genus, Geotrichum candidum, Giardia intestinalis, Gnathostoma spp, GSS prion, Guanarito virus, Haemophilus ducreyi, Haemophilus influenzae, Helicobacter pylori, Henipavirus (Hendra virus Nipah virus), Hepatitis A Virus, Hepatitis B Virus (HBV), Hepatitis C Virus (HCV), Hepatitis D Virus, Hepatitis E Virus, Herpes simplex virus 1 and 2 (HSV-1 and HSV- 2), Histoplasma capsulatum, HIV (Human immunodeficiency virus), Hortaea werneckii, Human bocavirus (HBoV), Human herpesvirus 6 (HHV-6) and Human herpesvirus 7 (HHV-7), Human metapneumovirus (hMPV), Human papillomavirus (HPV), Human parainfluenza viruses (HPIV), Japanese encephalitis virus, JC virus, Junin virus, Kingella kingae, Klebsiella granulomatis, Kuru prion, Lassa virus, Legionella pneumophila, Leishmania genus, Leptospira genus, Listeria monocytogenes, Lymphocytic choriomeningitis virus (LCMV), Machupo virus, Malassezia spp, Marburg virus, Measles virus, Metagonimus yokagawai, Microsporidia phylum, Molluscum contagiosum virus (MCV), Mumps virus, Mycobacterium leprae and Mycobacterium lepromatosis, Mycobacterium tuberculosis, Mycobacterium ulcerans, Mycoplasma pneumoniae, Naegleria fowleri, Necator americanus, Neisseria gonorrhoeae, Neisseria meningitidis, Nocardia asteroides, Nocardia spp, Onchocerca volvulus, Orientia tsutsugamushi, Orthomyxoviridae family (Influenza), Paracoccidioides brasiliensis, Paragonimus spp, Paragonimus westermani, ParvovirusBl 9, Pasteurella genus, Plasmodium genus, Pneumocystis jirovecii, Poliovirus, Rabies virus, Respiratory syncytial virus (RSV), Rhinovirus, rhinoviruses, Rickettsia akari, Rickettsia genus, Rickettsia prowazekii, Rickettsia rickettsii, Rickettsia typhi, Rift Valley fever virus, Rotavirus, Rubella virus, Sabia virus, Salmonella genus, Sarcoptes scabiei, SARS coronavirus, Schistosoma genus, Shigella genus, Sin Nombre virus, Hantavirus, Sporothrix schenckii, Staphylococcus genus, Staphylococcus genus, Streptococcus agalactiae, Streptococcus pneumoniae, Streptococcus pyogenes, Strongyloides stercoralis, Taenia genus, Taenia solium, Tick-borne encephalitis virus (TBEV), Toxocara canis or Toxocara cati, Toxoplasma gondii, Treponema pallidum, Trichinella spiralis, Trichomonas vaginalis, Trichophyton spp, Trichuris trichiura, Trypanosoma brucei, Trypanosoma cruzi, Ureaplasma urealyticum, Varicella zoster virus (VZV), V ariola major or Variola minor, vCJD prion, Venezuelan equine encephalitis virus, Vibrio cholerae, West Nile virus, Western equine encephalitis virus, Wuchereria bancrofti, Yellow fever virus, Yersinia enter ocolitica, Yersinia pestis, and Yersinia pseudotuberculosis.

[0193] The AP can comprise or can be derived from the full-length surface protein of a coronavirus. The disease or disorder can be an infectious disease or disorder caused by a coronavirus, and the AP can comprise or can be derived from an antigenic protein of a coronavirus. The term “coronavirus” as used herein refers to a virus in the family Coronaviridae , which is inturn classified within the order Nidovirales. The coronaviruses are large, enveloped, positive- stranded RNA viruses. The coronaviruses have the largest genomes of the RNA viruses known in the art and replicate by a unique mechanism that results in a high frequency of recombination. The coronaviruses include antigenic groups I, II, and III. Nonlimiting examples of coronaviruses include SARS coronavirus (e.g., SARS-CoV and SARS-CoV-2), MERS coronavirus, transmissible gastroenteritis virus (TGEV), human respiratory coronavirus, porcine respiratory coronavirus, canine coronavirus, feline enteric coronavirus, feline infectious peritonitis virus, rabbit coronavirus, murine hepatitis virus, sialodacryoadenitis virus, porcine hemagglutinating encephalomyelitis virus, bovine coronavirus, avian infectious bronchitis virus, and turkey coronavirus, as well as chimeras thereof. Additional information related to coronavirus including classification, virion structure, genome structure, genetics and pathology is described, for example, in KV Holmes, Encyclopedia of Virology, 1999: 291-298, the content of which is incorporated herein by reference.

[0194] In some embodiments, a coronavirus described herein is in the genus of Alphacoronavirus and the coronavirus antigens can be of or derived from any species or strains in the genus of Alpha-coronavirus . In some embodiments, a coronavirus described herein is in the genus of Beta-coronavirus and the coronavirus antigens can be of or derived from any species or strains in the genus of Beta-coronavirus . Member viruses in the genus of Alpha-coronavirus and Betacoronavirus are enveloped, positive-strand RNA viruses that can infect mammals.

[0195] A coronavirus described herein can be of any subgenus of Alpha-coronavirus genus, including but not limited to Colacovirus (e.g., Bat coronavirus CDPHE15), Decacovirus (e.g., Bat coronavirus HKU10 and Rhinolophus ferrumequinum alphacoronavirus HuB-2 13), Duvinacovirus (Human coronavirus 229E), Luchacovirus (e.g., Lucheng Rn rat coronavirus), Minacovirus (e.g., Mink coronavirus 1), Minunacovirus (e.g., Miniopterus bat coronavirus 1 and Miniopterus bat coronavirus HKU8), Myotacovirus (e.g., Myotis ricketti alphacoronavirus Sax- 2011), Nyctacovirus (e.g., Nyctalus velutinus alphacoronavirus SC-2013 and Pipistrellus kuhlii coronavirus 3398), Pedacovirus (e.g., Porcine epidemic diarrhea virus and Scotophilus bat coronavirus 512), Rhinacovirus (e.g., Rhinolophus bat coronavirus HKU2), Setracovirus (e.g., Human coronavirus NL63 and NL63-related bat coronavirus strain BtKYNL63-9b), Soracovirus (e.g., Sorex araneus coronavirus T14), Sunacovirus (e.g., Suncus murinus coronavirus X74), and Tegacovirus (e.g., Alphacoronavirus 1).

[0196] Within the genus Beta-coronavirus, five subgenera or lineages have been recognized, including Embecovirus (lineage A), Sarbecovirus (lineage B), Merbecovirus (lineage C), Nobecovirus (lineage D), and Hibecovirus. Accordingly, in some embodiments, a coronavirus described herein can be any strain or species in any of the subgenera or lineages of Beta-coronavirus.

[0197] For example, a coronavirus antigen can be of or derived from any species or strains in the subgenus of Embecovirus. including but not limited to Beta-coronavirus 1 (e.g., Bov / we coronavirus and human coronavirus OC43 . China Rattus coronavirus HKU24, Human coronavirus HKU1, Murine coronavirus (e.g., mouse hepatitis virus), and Myodes coronavirus 2JL14. The coronavirus antigen can be of or derived from any species or strains in the subgenus of Sarbecovirus. including but not limited to SARS-CoV, SARS-CoV2, 16BO133, Bat SARS CoV Rfl, Bat coronavirus HKU3 (BtCoV HKU3), LYRal 1, Bat SARS-CoV / Rp3, Bat SL-CoV YNLF 31C, Bat SL-CoV YNLF 34C, SHC014-CoV, WIV1, WIV16, Civet SARS-CoV, Rc- o319, SL-ZXC21, SL-ZC45, Pangolin SARSr-COV-GX, Pangolin SARSr-COV-GD, RshSTT182, RshSTT200, RacCS203, RmYN02, RpYN06, RaTG13, Bat CoV BtKY72, and Bat CoV BM48-31. The coronavirus antigen can be of any species or strains in the subgenus of Merbecovirus. including but not limited to Hedgehog coronavirus 7, MERS-CoV, Pipistrellus bat coronavirus HKU5, and Tylonycteris bat coronavirus HKU4. The coronavirus antigen can be of any species or strains in the subgenus of Nobecovirus, including but not limited to Eidolon bat coronavirus C704, Rousettus bat coronavirus GCCDC1, and Rousettus bat coronavirus HKU9. The coronavirus antigen can be of any species or strains in the subgenus of Hibecovirus. including but not limited to Bat Hp-betacoronavirus Zhejiang 2013.

[0198] The coronaviruses described herein can be, for example, phylogenetically clustered in functionally distinct clades. For example, the coronaviruses of lineage B Betacoronavirus (Sarbecovirus) can be clustered into clade 1, clade 2, clade 1 / 2, or clade 3 using the nucleotide sequences of nonstructural protein gene ORF la and ORF lb (see, for example, Hu et al., PLoS Pathog 13(11): el006698). Accordingly, the coronavirus antigens can be of or derived from any species or strain in any one of these clades. For example, the coronavirus antigens can be of any species or strain in clade 1, including but not limited to SARS-CoV, WIV1, LYRal 1, Rs7327, Rs4231, Rs4084, and SHC014. The coronavirus antigens can be of any species or strain in clade 2, including but not limited to As6526, Yunnan 2011, Shaanxi 2011, 279-2005, Rs4237, Rs4081, Rp3, Rs4247, HKU3-8, HKU3-13, GX2013, Longquan-140, YN2013, Rf4092, ZXC21, ZC45, JL2012, HuB2013, Rfl, HeB2013, and 273-2005. The coronavirus antigens can be of any species or strain in clade 1 / 2, including but not limited to SARS-CoV2. The coronavirus antigens can be of any species or strain in clade 3, including but not limited to BM48-31. The coronavirus antigen described herein can be of a coronavirus, for example, SARS, SARS-2, WIV1, SHC014, Rfl, RmYN02, pang 17, RaTG13, and Rs4081.

[0199] As exemplified herein, SARS virus (e.g., SARS-CoV and SARS-CoV-2) is an enveloped coronavirus carrying a single- stranded positive-sense RNA genome (~30 kb),belonging to the genus Betacoronavirus from the Coronaviridae family. The virus RNA encodes four structural proteins including spike (S), envelope (E), membrane (M), and nucleocapsid (N) proteins, 16 non- structural proteins, and nine accessory proteins. The S glycoprotein contains an ectodomain that can be processed into SI and S2 subunits, a transmembrane domain, and an intracellular domain. Both SARS-CoV and SARS-CoV-2 bind the human ACE2 via the receptor binding domain within the SI subunit to facilitate entry into host cells, followed by membrane fusion mediated by the S2 subunit.

[0200] A coronavirus antigen of a coronavirus herein described can be any of a variety of coronavirus proteins capable of inducing an immune response against a coronavirus. Suitable coronavirus antigens are those that can elicit a protective immune response, such as producing broadly neutralizing antibodies. For example, the coronavirus antigen can comprise a coronavirus spike (S) protein, spike receptor binding domain (RBD), SI subunit, S2 subunit, spike full ectodomain proteins, papain-like proteases, 3 CL proteases, nucleocapsid proteins, envelope proteins, membrane proteins, or any of the structural, non- structural or accessory proteins that form a coronavirus.

[0201] In some embodiments, a coronavirus antigen used herein comprises a spike (S) protein or a portion thereof. A S protein is one of four major structural proteins covering the surface of each virion. The S protein, comprising a SI subunit and a S2 subunit, is a highly glycosylated, type I transmembrane protein capable of binding to a host-cell receptor and mediates viral entry. The S protein comprises a domain referred to as the RBD that mediates the interaction with the host-cell receptor to enter the host cell after one or more RBDs adopts an “up” position to bind the host receptor. It is believed that after binding the receptor, a nearby host protease cleaves the spike, which releases the spike fusion peptide, facilitating virus entry. Known host receptors for coronaviruses (e.g., Beta-coronaviruses) include antiotensin-converting enzyme 2 (ACE2), dipeptidyl peptidase-4 (DPP4) or sialic acids. For example, the RBDs of human coronaviruses SARS-CoV-2, SARS-CoV, HCoV-NL63, and related animal coronaviruses (WIV1 and SCH014) use ACE2 as their host receptor, while MERS-CoV uses DPP4 as its host receptor.

[0202] The coronavirus antigen used herein can, for example, comprise a coronavirus nucleocapsid protein (N protein) or a portion thereof. The N protein is a multifunctional RNA- binding protein required for viral RNA transcription, replication, and packaging. The N protein consists of three domains, an N-terminal RNA-binding domain, a central intrinsically disordered region, followed by a C-terminal dimerization domain. The RNA-binding domain contains multiple positively charged binding surfaces that form charged interactions with RNA promoting its helical arrangement. The coronavirus antigen used herein can comprise any of these N protein domains or a portion thereof.

[0203] In some embodiments, the coronavirus antigen used herein comprises a coronavirus membrane protein (M protein) or a portion thereof. The M protein is the most abundant structural protein and defines the shape of the viral envelope. The M protein is regarded as the central organizer of the viral assembly, interacting with other major coronaviral structural proteins.

[0204] In some embodiments, the coronavirus antigen used herein comprises a coronavirus envelope protein (E protein) or a portion thereof. The E protein is a small membrane protein and minor component of the virus particles. Without being bound to any theory, it is believed that the E protein plays roles in virion assembly and morphogenesis, alteration of the membrane of host cells and virus-host cell interaction.

[0205] In some embodiments, the coronavirus antigen used herein comprises a coronavirus hemagglutinin-esterase protein (HE protein) or a portion thereof. The HE protein, which is another envelope protein, mediates reversible attachment to O-acetylated sialic acids by acting both as lectins and receptor-destroying enzymes.

[0206] In some embodiments, the coronavirus antigen used herein comprises a coronavirus papain-like protease or a portion thereof. The coronavirus papain-like protease is one of several nonstructural proteins, and is responsible for processing of viral proteins into functional, mature subunits during maturation. For example, the coronavirus papain-like protease can cleave a site at the amino-terminal end of the viral replicase region. In addition to its role in viral protein maturation, papain-like protease exhibits both a deubiquitinating and deISG15ylating activity. In vivo, this protease antagonizes innate immunity by acting on IFN beta and NF- kappa B signaling pathways.

[0207] In some embodiments, the coronavirus antigen used herein comprises a coronavirus 3CL protease or a portion thereof. The 3CL protease is another main protease in addition to the papain-like protease and is required for processing of viral polypeptides into distinct, functional proteins. In some embodiments, the 3CL protease is a SARS-CoV-2 3CL Protease, which is a C30-type cysteine protease located within the non- structural proteins 3 (NS3) region of the viral polypeptide. Analysis of the Coronavirus genome reveals at least 11 sites of cleavage for the 3CL protease, many containing the amino acid sequence LQ[S / A / G],

[0208] The coronavirus antigen disclosed herein can, in some embodiments, comprise a S protein or a portion thereof, a N protein or a portion thereof, a HE protein or a portion thereof, a papain-like protease or a portion thereof, a coronavirus 3 CL protease or a portion thereof, a M protein or a portion thereof, or a combination thereof.

[0209] In some embodiments, the coronavirus antigen can be an immunogenic portion of a coronavirus protein herein described. It will be appreciated by those skilled in the art that animmunogenic portion of a coronavirus antigen can be fragments of the S protein (e.g., spike protein RBD), N protein, HE protein, papain-like protease, 3 CL protease, or M protein capable of eliciting an immune response against one or more coronaviruses. The immunogenic portion can comprise about, at least or at least about, at most or at most about, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, or a number or a range between any two of these values, contiguous amino acid residues from the coronavirus proteins. In some embodiments, the immunogenic portion comprises a S protein RBD or a portion thereof. The portion of the S protein RBD can comprise the receptor binding motif of the S protein RBD.

[0210] One or more of the plurality of CoV antigens can be of Co Vs in the genus of Alpha-CoV and / or Beta-CoV, and optionally each of the plurality of CoV antigens are of CoVs in the genus of Beta-CoV. The plurality of CoV antigens can be of CoVs in the subgenus of Sarbecovirus. The first CoV and the second CoV can be in the genus of Beta-CoV, optionally in the subgenus of Sarbecovirus. The plurality of CoV antigens can be of CoVs selected from the group consisting of: SARS-CoV, SARS-CoV-2, WIV1, SHC014, Rfl, RmYN02, pangl7, RaTG13, Rs4081, LYRal 1, HKU3, Yunnan2011, BtKY72, BM48-31, WIV16, Khosta-1, and Khosta-2. The first CoV, the second CoV, or both can be selected from the group consisting of: SARS-CoV, SARS-CoV-2, WIV1, SHC014, Rfl, RmYN02, pangl7, RaTG13, Rs4081, LYRal 1, HKU3, Yunnan2011, BtKY72, BM48-31, WIV16, Khosta-1, and Khosta-2. The CoV can be selected from a species or subspecies of SARS-CoV, SARS-CoV- 1, SARS-CoV-2, MERS-CoV, SL-CoV-WIVl, HKU4, HKU5, HCoV-OC43, HCoV-HKUl, HKU9, HKU3, HKU8, HKU24, NL63, SHC014, 229E and / or SARS-CoV-2 variants B.1.351, B.l.1.7, P. l, B.1.617.2, B.1.1.529, BA.1, BA.1.1, BA.2, BA.3, BA.4, BA.5 and other descendent lineages.

[0211] The CoV can be selected from a species or subspecies of Embecovirus, Sarbecovirus, Merbecovirus, Nobevovirus, Hibecovirus, SARSr-CoV, MERS-CoV, or any combination thereof. The CoV can be selected from a beta-CoV from the sarbe-, embeco-, merbeco-, and / or nobecovirus lineages. The CoV can be selected from a sarbecovirus strain, optionally, SARS, LYRal 1, Rfl, Rs4081, BtKY72, and BM48-31. The CoV can be selected from a merbecovirus strain, optionally HKU4, HKU5, HKU25, BtCoV-Vs-CoVl, MERS-related NL13845, and MERS-related NL140422. The CoV can be selected from an embecovirus strain, optionally HKU1, Rat CoV Parker, PHEV, Equine CoV, Rodent CoV, and Longquan Rat CoV.Tumor-associated Antisens, Autoimmune Antisens, and Allergenic Antisens

[0212] The disease or disorder can be a disease associated with expression of a tumor- associated antigen, and the antigenic protein can be a tumor-associated antigen. A tumor- associated antigen can be a tumor-specific antigen. In some embodiments, the tumor-associated antigen is selected from the group comprising: lA01_HLA-A / m (UniProtKB: P30443); 1A02(UniProtKB: P01892); 5T4 (UniProtKB: Q13641); ACRBP (UniProtKB: Q8NEB7); AFP (UniProtKB: P02771); AKAP4 (UniProtKB: Q5JQC9); alpha-actinin-_4 / m (UniProtKB: B4DSX0); alpha-actinin-_4 / m (UniProtKB: B4E337); alpha-actinin-_4 / m (UniProtKB: 043707); alpha-methylacyl-coenzyme_A_racemase (UniProtKB: A0A024RE16); alpha-methylacyl- coenzyme A racemase (UniProtKB: A8KAC3); ANDR (UniProtKB: P10275); ART-4 (UniProtKB: Q9ULX3); ARTCl / m (UniProtKB: P52961); AURKB (UniProtKB: Q96GD4); B2MG (UniProtKB: P6176...

Claims

WHAT IS CLAIMED IS:

1. A composition, comprising: a nucleic acid composition comprising:(i) a polynucleotide encoding a dimerization fusion protein, wherein the dimerization fusion protein comprises an optional antigenic polypeptide (AP) and a heterologous cytoplasmic tail, and(ii) a polynucleotide encoding an adapter fusion protein comprising an endosomal sorting complex required for transport (ESCRT)-recruiting domain (ERD) and an adapter domain capable of binding the heterologous cytoplasmic tail to form a heterodimer, wherein binding of the adapter domain to the heterologous cytoplasmic tail is capable of recruiting one or more ESCRT proteins to the heterodimer, thereby inducing a plurality of dimerization fusion proteins to self-assemble into an enveloped nanoparticle (ENP) secreted from a cell in which the dimerization fusion protein and adapter fusion protein are expressed, thereby generating a population of ENPs.

2. A composition, comprising: a nucleic acid composition comprising(i) n polynucleotides each encoding an nth dimerization fusion protein, wherein n is an integer from 2 to 500, wherein each dimerization fusion protein comprises an optional antigenic polypeptide (AP) and a heterologous cytoplasmic tail, wherein at least two of the dimerization fusion proteins differ with respect to the AP; and(ii) a polynucleotide encoding an adapter fusion protein comprising an endosomal sorting complex required for transport (ESCRT)-recruiting domain (ERD) and an adapter domain capable of binding the heterologous cytoplasmic tail to form a heterodimer, wherein binding of the adapter domain to the heterologous cytoplasmic tail is capable of recruiting one or more ESCRT proteins to the heterodimer, thereby inducing a plurality of dimerization fusion proteins to self-assemble into an enveloped nanoparticle (ENP) secreted from a cell in which the n dimerization fusion proteins and the adapter fusion protein are expressed, thereby generating a population of ENPs.

3. The composition of any one of claims 1-2, wherein the plurality of dimerization fusion proteins are capable of being presented on the surface of the cell in which the plurality of dimerization fusion proteins are expressed, optionally wherein the cell is: a cell of a subject;an in vivo cell, an ex vivo cell, or an in situ cell; and / or an adherent cell or a suspension cell.

4. The composition of any one of claims 1-3, wherein the self-assembly of an ENP: does not require an exogenous nucleic acid other than the nucleic acid composition, and / or does not require any exogenous components other than the dimerization fusion protein and the adapter fusion protein.

5. The composition of any one of claims 1-4, wherein upon secretion from a cell of a subject, the ENPs are capable of distributing within one or more tissues of the subject, optionally the one or more tissues comprise adrenal gland tissue, appendix tissue, bladder tissue, bone, bowel tissue, brain tissue, breast tissue, bronchi, coronal tissue, ear tissue, esophagus tissue, eye tissue, gall bladder tissue, genital tissue, heart tissue, hypothalamus tissue, kidney tissue, large intestine tissue, intestinal tissue, larynx tissue, liver tissue, lung tissue, lymph nodes, mouth tissue, nose tissue, pancreatic tissue, parathyroid gland tissue, pituitary gland tissue, prostate tissue, rectal tissue, salivary gland tissue, skeletal muscle tissue, skin tissue, small intestine tissue, spinal cord, spleen tissue, stomach tissue, thymus gland tissue, trachea tissue, thyroid tissue, ureter tissue, urethra tissue, soft and connective tissue, peritoneal tissue, blood vessel tissue, fat tissue, or any combination thereof, further optionally the ENPs engage a plurality of immune cells in the one or more tissues, thereby mimicking a natural infection.

6. A composition, comprising: a population of enveloped nanoparticles (ENPs), wherein each of the ENPs comprises a plurality of dimerization fusion proteins each comprising a heterologous cytoplasmic tail and optionally, an antigenic polypeptide (AP).

7. The composition of any one of claims 1-6, wherein the ENPs are derived from expression of the nucleic acid composition of any one of claims 1-5.

8. The composition of any one of claims 1-7, wherein the adapter fusion protein comprises, from N-terminus to C-terminus: the adapter domain, an optional linker, and the ERD, optionally the linker: is a flexible linker, a rigid linker, or a hybrid linker; is hydrophilic or hydrophobic; is between 1 and 250 amino acids; comprises one or more flexible amino acid residues, optionally about 1 to about 250flexible amino acid residues, further optionally the flexible amino acid residues comprise glycine, serine, or a combination thereof; and / or comprises 3 repeating amino acid subunits or more.

9. The composition of any one of claims 1-8, wherein the heterologous cytoplasmic tail is derived from or comprises a cytoplasmic tail of a cell surface protein, and wherein the adapter domain is (1) capable of binding the heterologous cytoplasmic tail derived from or comprising the cytoplasmic tail of said cell surface protein and (2) capable of targeting the adapter fusion protein to the plasma membrane, optionally the cell surface protein is or is derived from a human protein, a nonhuman mammalian protein, an avian protein, or a reptile protein and / or the adapter domain is or is derived from a human protein, a non-human mammalian protein, an avian protein, or a reptile protein, optionally the heterologous cytoplasmic tail comprises or is derived from a cytoplasmic tail (CT) of CD4 or CD8 and / or the adapter domain comprises or is derived from at least a portion of Lek tyrosine kinase, further optionally: the CD4 CT comprises the sequence of SEQ ID NO: 95 or SEQ ID NO: 96; the CD8 CT comprises the sequence of SEQ ID NO: 101; the adapter domain comprises the sequence of SEQ ID NO: 43 or SEQ ID NO: 97; and / or the heterologous cytoplasmic tail of CD4 comprises the sequence of SEQ ID NO: 98, SEQ ID NO: 99, or SEQ ID NO: 100.

10. The composition of claim 9, wherein: a) the heterologous cytoplasmic tail of CD4 comprises the sequence of SEQ ID NO: 102, and the adapter domain comprises the sequence of SEQ ID NO: 105; b) the heterologous cytoplasmic tail of CD4 comprises the sequence of SEQ ID NO: 103, and the adapter domain comprises the sequence of SEQ ID NO: 106; or c) the heterologous cytoplasmic tail of CD4 comprises the sequence of SEQ ID NO: 104, and the adapter domain comprises the sequence of SEQ ID NO: 107.

11. The composition of any one of claims 9-10, wherein the composition further comprises: iii) a polynucleotide encoding a second adapter fusion protein comprising an endosomal sorting complex required for transport (ESCRT)-recruiting domain (ERD) and an adapter domain capable of binding the heterologous cytoplasmic tail to form a heterodimer, wherein the adapter domain of the second adapter fusion protein is the same or different from the adapter domain ofthe adapter fusion protein, and wherein the ERD of the second adapter fusion protein is the same or different from the ERD of the adapter fusion protein, optionally wherein the adapter domain of the second adapter fusion protein comprises the sequence of SEQ ID NO: 109 and the adapter domain of the adapter fusion protein comprises the sequence of SEQ ID NO: 43 or SEQ ID NO: 97.

12. The composition of claim 9, wherein:(i) the heterologous cytoplasmic tail comprises or is derived from G. gallus CD4 CT, optionally comprising the sequence of SEQ ID NO: 110, and wherein the adapter domain comprises or is derived from at least a portion of G. gallus Lek tyrosine kinase, optionally comprising the sequence of SEQ ID NO: 113;(ii) wherein the heterologous cytoplasmic tail comprises or is derived from P. vitticeps CD4 CT, optionally comprising the sequence of SEQ ID NO: 111;(iii) wherein the adapter domain comprises or is derived from at least a portion of P. vitticeps Lek tyrosine kinase, optionally comprising the sequence of SEQ ID NO: 114; or(iv) wherein the heterologous cytoplasmic tail comprises or is derived from N scutatus CD4 CT, optionally comprising the sequence of SEQ ID NO: 112, and wherein the adapter domain comprises or is derived from at least a portion of N scutatus Lek tyrosine kinase, optionally comprising the sequence of SEQ ID NO: 115.

13. The composition of any one of claims 9-12, wherein: the dimerization fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 41-42, 45, 48-59, 63, 68-69, and 83-85; and / or wherein the adapter fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 44, 46, 60-62, 64-65, 70-82, and 86-88.

14. The composition of any one of claims 9-12, wherein:(i) the dimerization fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 51; and(ii) the adapter fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 44, 64, 77, and 80-82.

15. The composition of any one of claims 9-12, wherein:(i) the dimerization fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to thesequence of SEQ ID NO: 51, and (ii) the adapter fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 44;(i) the dimerization fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 51, and (ii) the adapter fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 64;(i) the dimerization fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 51, and (ii) the adapter fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 77;(i) the dimerization fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 51, and (ii) the adapter fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 80;(i) the dimerization fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 51, and (ii) the adapter fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 81;(i) the dimerization fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 51, and (ii) the adapter fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 82;(i) the dimerization fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 69, and (ii) the adapter fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 44, 64, 77, and 80-82;(i) the dimerization fusion protein comprises an amino acid sequence having atleast 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 69, and (ii) the adapter fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 44;(i) the dimerization fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 69, and (ii) the adapter fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 64;(i) the dimerization fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 69, and (ii) the adapter fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 77;(i) the dimerization fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 69, and (ii) the adapter fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 80;(i) the dimerization fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 69, and (ii) the adapter fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 81; or(i) the dimerization fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 69, and (ii) the adapter fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 82.

16. The composition of any one of claims 1-7, wherein: the dimerization fusion protein comprises, (i) from N-terminus to C-terminus: the antigenic polypeptide, an optional linker, and the heterologous cytoplasmic tail, optionally wherein the dimerization fusion protein comprises an endogenous cytoplasmic tail N-terminal to the heterologous cytoplasmic tail or (ii) from C-terminus to N-terminus: the antigenic polypeptide, an optional linker, and the heterologous cytoplasmic tail, optionally wherein the dimerizationfusion protein comprises an endogenous cytoplasmic tail C-terminal to the heterologous cytoplasmic tail; and / or wherein the adapter fusion protein comprises, from N-terminus to C-terminus: a heterologous membrane-targeting domain, an optional first linker, the adapter domain, an optional second linker, and the ERD, optionally the linker, the first linker, and / or the second linker: is a flexible linker, a rigid linker, or a hybrid linker; is hydrophilic or hydrophobic; is between 1 and 250 amino acids; comprises one or more flexible amino acid residues, optionally about 1 to about 250 flexible amino acid residues, further optionally the flexible amino acid residues comprise glycine, serine, or a combination thereof; and / or comprises 3 repeating amino acid subunits or more.

17. The composition of claim 16, wherein the heterologous membrane targeting domain comprises or is derived from at least a portion of LAT, PAG, LCK, FYN, LAX, CD2, CD3, CD4, CD5, CD7, CD8a, PD1, SRC, or LYN, optionally the heterologous membrane targeting domain comprises or is derived from FYN, further optionally the heterologous membrane targeting domain comprises the sequence of SEQ ID NO: 118.

18. The composition of any one of claims 16-17, wherein the heterologous cytoplasmic tail and / or the adapter domain are each selected from the group comprising DHD9 heterodimer a, DHD13 XAAA heterodimer a, DHD13 XAXA heterodimer a, DHD13 XAAX heterodimer a, DHD13_2:341 heterodimer a, DHD13 AAAA heterodimer a, DHD13 BAAA heterodimer a, DHD13_4: 123 heterodimer a, DHD13 E234 heterodimer a, DHD15 heterodimer a, DHD20 heterodimer a, DHD21 heterodimer a, DHD25 heterodimer a, DHD27 heterodimer a, DHD30 heterodimer a, DHD33 heterodimer a, DHD34 XAAXA heterodimer a, DHD34 XAXXA heterodimer a, DHD34 XAAAA heterodimer a, DHD36 heterodimer a, DHD37 ABXB heterodimer a, DHD37 BBBB heterodimer a, DHD37_XBXB heterodimer a, DHD37 AXXB heterodimer a, DHD37_3: 124 heterodimer a, DHD37 E234 heterodimer a, DHD37 AXBB heterodimer a, DHD37 XBBA heterodimer a, DHD39 heterodimer a, DHD40 heterodimer a, DHD43 heterodimer a, DHD65 heterodimer a, DHD70 heterodimer a, DHD88 heterodimer a, DHD89 heterodimer a, DHD90 heterodimer a, DHD91 heterodimer a, DHD92 heterodimer a, DHD93 heterodimer a, DHD94 heterodimer a, DHD94_3:214 heterodimer a, DHD94_2: 143 heterodimer a, DHD95 heterodimer a, DHD96 heterodimer a, DHD97 heterodimer a, DHD98 heterodimer a, DHD99 heterodimer a, DHD100 heterodimer a, DHD101 heterodimer a, DHD102 heterodimer a, DHD102_l :243 heterodimer a, DHD103 heterodimer a, DHD103_l :423heterodimer a, DHD104 heterodimer a, DHD105 heterodimer a, DHD106 heterodimer a, DHD107 heterodimer a, DHD108 heterodimer a, DHD109 heterodimer a, DHD110 heterodimer a, DHD111 heterodimer a, DHD112 heterodimer a, DHD113 heterodimer a, DHD114 heterodimer a, DHD115 heterodimer a, DHD116 heterodimer a, DHD117 heterodimer a, DHD118 heterodimer a, DHD119 heterodimer a, DHD120 heterodimer a, DHD121 heterodimer a, DHD122 heterodimer a, DHD123 heterodimer a, DHD124 heterodimer a, DHD125 heterodimer a, DHD126 heterodimer a, DHD127 heterodimer a, DHD128 heterodimer a, DHD129 heterodimer a, DHD130 heterodimer a, DHD145 heterodimer a, DHD146 heterodimer a, DHD147 heterodimer a, DHD1 heterodimer a, DHD2 heterodimer a, DHD3 heterodimer a, DHD4 heterodimer a, DHD5 heterodimer a, DHD6 heterodimer a, DHD7 heterodimer a, DHD8 heterodimer a, DHD16 heterodimer a, DHD18 heterodimer a, DHD19 heterodimer a, DHD22 heterodimer a, DHD23 heterodimer a, DHD24 heterodimer a, DHD26 heterodimer a, DHD28 heterodimer a, DHD29 heterodimer a, DHD31 heterodimer a, DHD32 heterodimer a, DHD38 heterodimer a, DHD60 heterodimer a, DHD63 heterodimer a, DHD66 heterodimer a, DHD67 heterodimer a, DHD69 heterodimer a, DHD71 heterodimer a, DHD72 heterodimer a, DHD73 heterodimer a, DHD148 heterodimer a, DHD149 heterodimer a, DHD150 heterodimer a, DHD151 heterodimer a, DHD152 heterodimer a, DHD153 heterodimer a, DHD154 heterodimer a, DHD155 heterodimer a, DHD156 heterodimer a, DHD157 heterodimer a, DHD158 heterodimer a, DHD159 heterodimer a, DHD160 heterodimer a, DHD161 heterodimer a, DHD162 heterodimer a, DHD163 heterodimer a, DHD164 heterodimer a, DHD165 heterodimer a, DHD166 heterodimer a, DHS17 heterodimer a, DHD17 heterodimer a, DHD131 heterodimer a, DHD132 heterodimer a, DHD133 heterodimer a, DHD134 heterodimer a, DHD135 heterodimer a, DHD136 heterodimer a, DHD137 heterodimer a, DHD138 heterodimer a, DHD139 heterodimer a, DHD140 heterodimer a, DHD141 heterodimer a, DHD142 heterodimer a, DHD143 heterodimer a, DHD144 heterodimer a, DHD9 heterodimer b, DHD13 XAAA heterodimer b, DHD13 XAXA heterodimer b, DHD13 XAAX heterodimer b, DHD13_2:341 heterodimer b, DHD13 AAAA heterodimer b, DHD13 BAAA heterodimer b, DHD13_4: 123 heterodimer b, DHD13_1 :234 heterodimer b, DHD15 heterodimer b, DHD20 heterodimer b, DHD21 heterodimer b, DHD25 heterodimer b, DHD27 heterodimer b, DHD30 heterodimer b, DHD33 heterodimer b, DHD34 XAAXA heterodimer b, DHD34 XAXXA heterodimer b, DHD34 XAAAA heterodimer b, DHD36 heterodimer b, DHD37 ABXB heterodimer b, DHD37 BBBB heterodimer b, DHD37 XBXB heterodimer b, DHD37 AXXB heterodimer b, DHD37_3: 124 heterodimer b, DHD37_1 :234 heterodimer b, DHD37 AXBB heterodimer b, DHD37 XBBA heterodimer b, DHD39 heterodimer b, DHD40 heterodimer b, DHD43 heterodimer b, DHD65 heterodimer b, DHD70 heterodimer b, DHD88 heterodimer b, DHD89heterodimer b, DHD90 heterodimer b, DHD91 heterodimer b, DHD92 heterodimer b, DHD93 heterodimer b, DHD94 heterodimer b, DHD94_3:214 heterodimer b, DHD94_2: 143 heterodimer b, DHD95 heterodimer b, DHD96 heterodimer b, DHD97 heterodimer b, DHD98 heterodimer b, DHD99 heterodimer b, DHD100 heterodimer b, DHD101 heterodimer b, DHD102 heterodimer b, DHD102_l :243 heterodimer b, DHD103 heterodimer b, DHD103_l :423 heterodimer b, DHD104 heterodimer b, DHD105 heterodimer b, DHD106 heterodimer b, DHD107 heterodimer b, DHD108 heterodimer b, DHD109 heterodimer b, DHD110 heterodimer b, DHD111 heterodimer b, DHD112 heterodimer b, DHD113 heterodimer b, DHD114 heterodimer b, DHD1 15 heterodimer b, DHD116 heterodimer b, DHD117 heterodimer b, DHD118 heterodimer b, DHD119 heterodimer b, DHD120 heterodimer b, DHD121 heterodimer b, DHD122 heterodimer b, DHD123 heterodimer b, DHD124 heterodimer b, DHD125 heterodimer b, DHD126 heterodimer b, DHD127 heterodimer b, DHD128 heterodimer b, DHD129 heterodimer b, DHD130 heterodimer b, DHD145 heterodimer b, DHD146 heterodimer b, DHD147 heterodimer b, DHD1 heterodimer b, DHD2 heterodimer b, DHD3 heterodimer b, DHD4 heterodimer b, DHD5 heterodimer b, DHD6 heterodimer b, DHD7 heterodimer b, DHD8 heterodimer b, DHD16 heterodimer b, DHD18 heterodimer b, DHD19 heterodimer b, DHD22 heterodimer b, DHD23 heterodimer b, DHD24 heterodimer b, DHD26 heterodimer b, DHD28 heterodimer b, DHD29 heterodimer b, DHD31 heterodimer b, DHD32 heterodimer b, DHD38 heterodimer b, DHD60 heterodimer b, DHD63 heterodimer b, DHD66 heterodimer b, DHD67 heterodimer b, DHD69 heterodimer b, DHD71 heterodimer b, DHD72 heterodimer b, DHD73 heterodimer b, DHD148 heterodimer b, DHD149 heterodimer b, DHD150 heterodimer b, DHD15 1 heterodimer b, DHD152 heterodimer b, DHD153 heterodimer b, DHD154 heterodimer b, DHD155 heterodimer b, DHD156 heterodimer b, DHD157 heterodimer b, DHD158 heterodimer b, DHD159 heterodimer b, DHD160 heterodimer b, DHD161 heterodimer b, DHD162 heterodimer b, DHD163 heterodimer b, DHD164 heterodimer b, DHD165 heterodimer b, DHD166 heterodimer b, DHS17 heterodimer b, DHD17 heterodimer b, DHD131 heterodimer b, DHD132 heterodimer b, DHD133 heterodimer b, DHD134 heterodimer b, DHD135 heterodimer b, DHD136 heterodimer b, DHD137 heterodimer b, DHD138 heterodimer b, DHD139 heterodimer b, DHD140 heterodimer b, DHD141 heterodimer b, DHD142 heterodimer b, DHD143 heterodimer b, DHD144 heterodimer b, portions thereof, derivatives thereof, or any combination thereof.

19. The composition of any one of claims 16-17, wherein the heterologous cytoplasmic tail and / or the adapter domain comprises or is derived from SYNZIP1, SYNZIP2, SYNZIP3, SYNZIP4, SYNZIP5, SYNZIP6, SYNZIP7, SYNZIP8, SYNZIP9, SYNZIP10, SYNZIP11, SYNZIP12, SYNZIP13, SYNZIP14, SYNZIP15, SYNZIP16, SYNZIP17, SYNZIP18,SYNZIP19, SYNZIP20, SYNZIP21, SYNZIP22, SYNZIP23, BATF, FOS, ATF4, BACH1, JUND, NFE2L3, AZip, BZip, a PDZ domain ligand, an SH3 domain, a PDZ domain, a GTPase binding domain, a leucine zipper domain, an SH2 domain, a PTB domain, an FHA domain, a WW domain, a 14-3-3 domain, a death domain, a caspase recruitment domain, a bromodomain, a chromatin organization modifier, a shadow chromo domain, an F-box domain, a HECT domain, a RING finger domain, a sterile alpha motif domain, a glycine-tyrosine-phenylalanine domain, a SNAP domain, a VHS domain, an ANK repeat, an armadillo repeat, a WD40 repeat, an MH2 domain, a calponin homology domain, a Dbl homology domain, a gelsolin homology domain, a PB1 domain, a SOCS box, an RGS domain, a Toll / IL-1 receptor domain, a tetratricopeptide repeat, a TRAF domain, a Bcl-2 homology domain, a coiled-coil domain, a bZIP domain, portions thereof, variants thereof, or any combination thereof.

20. The composition of any one of claims 16-17, wherein: the heterologous cytoplasmic tail comprises or is derived from ACIDpl or BASEp 1 or wherein the heterologous cytoplasmic tail comprises or is derived from N5 or N6, optionally the heterologous cytoplasmic tail comprises the sequence of any one of SEQ ID NOs: 116-117 and 121-122; and / or wherein the adapter domain comprises or is derived from ACIDpl or BASEp 1 or wherein the heterologous cytoplasmic tail comprises or is derived from N5 or N6, optionally the adapter domain comprises the sequence of any one of SEQ ID NOs: 116-117 and 121-122.

21. The composition of claim 20, wherein: the dimerization fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 89-90 and 93; and / or wherein the adapter fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 91-92 and 94.

22. The composition of claim 20, wherein(i) the dimerization fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 93; and(ii) the adapter fusion protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 94.

23. The composition of any one of claims 1-22, wherein (i) the polynucleotide encoding the dimerization fusion protein or the n polynucleotides each encoding an nthdimerization fusion protein, and (ii) the polynucleotide encoding the adapter fusion protein, are each present in a different nucleic acid molecule, optionally wherein the amount of(i) the polynucleotide encoding the dimerization fusion protein or the n polynucleotides each encoding an nth dimerization fusion protein; and(ii) the polynucleotide encoding the adapter fusion protein, are present in the composition at a molar ratio of about 9: 1, 5: 1, or 1 : 1.

24. The composition of any one of claims 1-22, wherein (i) the polynucleotide encoding the dimerization fusion protein or the n polynucleotides each encoding an nth dimerization fusion protein, and (ii) the polynucleotide encoding the adapter fusion protein, are present in the same nucleic acid molecule.

25. The composition of any one of claims 1-24, wherein: the cell expressing the dimerization fusion protein or the plurality of dimerization fusion proteins and the adapter fusion protein exhibits at least a 2-fold increase in AP expression at the surface of the cell, relative to a cell expressing a fusion protein or a plurality of fusion proteins comprising an AP and an ERD and does not express the adapter fusion protein; and / or wherein the production of ENPs from the cell expressing the dimerization fusion protein or the plurality of dimerization fusion proteins and the adapter fusion protein is increased by at least 2-fold, relative to a cell expressing a fusion protein or a plurality of fusion proteins comprising an AP and an ERD and does not express the adapter fusion protein.

26. The composition of any one of claims 1-25, wherein the nucleic acid composition further comprises a polynucleotide comprising or encoding a tetherin inhibitor, optionally the tetherin inhibitor is capable of modulating expression, concentration, localization, stability, and / or wherein activity of tetherin and / or wherein presence or expression of the tetherin inhibitor in the cell results in an increase in ENP production by the cell by at least 2-fold, relative to a cell that does not comprise or express the tetherin inhibitor, further optionally the tetherin inhibitor comprises a dsRNA, an siRNA, an shRNA, a pre- miRNA, a pri-miRNA, a miRNA, an stRNA, an IncRNA, a piRNA, a snoRNA, or a protein.

27. The composition of claim 26, wherein one or more of (i) the polynucleotide comprising or encoding the tetherin inhibitor, (ii) the polynucleotide encoding the dimerization fusion protein or the n polynucleotides each encoding an nth dimerization fusion protein, and (iii) the polynucleotide encoding the adapter fusion protein, are present in a same or a different nucleic acid molecule,optionally the amount of(i) the polynucleotide comprising or encoding the tetherin inhibitor; and(ii) the polynucleotide encoding the dimerization fusion protein or the n polynucleotides each encoding an nth dimerization fusion protein, and / or the polynucleotide encoding the adapter fusion protein, are present in the composition at a molar ratio of about 1 : 1, 1 :5 or 1 :25.

28. The composition of any one of claims 26-27, wherein the polynucleotide comprising or encoding the tetherin inhibitor and the polynucleotide encoding the dimerization fusion protein are present in the same nucleic acid or wherein the polynucleotide comprising or encoding the tetherin inhibitor and the polynucleotide encoding the adapter fusion protein are present in the same nucleic acid.

29. The composition of any one of claims 26-28, wherein the tetherin inhibitor comprises or is derived from a viral protein, optionally the virus is HIV-1, HIV-2, SIV, Ebola virus, KSHV, SARS CoV, or SARS-CoV-2, further optionally the tetherin inhibitor comprises HIV-1 Vpu protein, KSHV K5 protein, SARS-CoV-2 ORF7a, HIV-2 Env, Ebola GP, SIV Env, SIV Vpu, SIV Nef, or any portions, variants or derivatives thereof.

30. The composition of any one of claims 26-29, wherein the tetherin inhibitor comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 132-134, optionally the tetherin inhibitor comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 132.

31. A composition, comprising: a nucleic acid composition comprising:(i) a polynucleotide encoding a recombinant protein comprising an optional antigenic polypeptide (AP) and a transmembrane domain, and(ii) a polynucleotide encoding a chimeric protein comprising: a) a signal peptide, b) a cell surface domain, c) a transmembrane domain, and d) a cytoplasmic domain; wherein the recombinant protein and / or the chimeric protein further comprises an endosomal sorting complex required for transport (ESCRT)-recruiting domain (ERD), optionally the recombinant protein further comprises a cytoplasmic domain; and wherein the recombinant protein and the chimeric protein are capable of co-localizing to a site on the plasma membrane of the cell, and wherein the ERD is capable of recruiting one or more ESCRT proteins to the site on the plasma membrane, thereby inducing a plurality of recombinant proteins and chimeric proteins to self-assemble into an enveloped nanoparticle (ENP) secreted from a cell in which the recombinant protein and chimeric protein are expressed, thereby generating a population of ENPs, optionally the cytoplasmic domain of the recombinant protein and the cytoplasmic domain of the chimeric protein are capable of co-localizing the recombinant protein and the chimeric protein to a site on the plasma membrane of the cell, optionally the transmembrane domain of the recombinant protein and the transmembrane domain of the chimeric protein are capable of co-localizing the recombinant protein and the chimeric protein to a site on the plasma membrane of the cell.

32. A composition, comprising: a nucleic acid composition comprising:(i) n polynucleotides each encoding a recombinant protein comprising an optional antigenic polypeptide (AP) and a transmembrane domain, wherein at least two of the recombinant proteins differ with respect to the AP, wherein n is an integer from 2 to 500; and(ii) z polynucleotides each encoding a chimeric protein comprising: a) a signal peptide, b) a cell surface domain, wherein at least two of the chimeric proteins differ with respect to the cell surface domain, c) a transmembrane domain, and d) a cytoplasmic domain wherein z is an integer from 1 to 500, wherein the nth recombinant proteins and / or the zth chimeric proteins further comprise an endosomal sorting complex required for transport (ESCRT)-recruiting domain (ERD), optionally the nth recombinant proteins further comprise a cytoplasmic domain, and wherein each of the nth recombinant protein and each of the zth chimeric protein are capable of co-localizing to a site on the plasma membrane of the cell, and the ERD is capable of recruiting one or more ESCRT proteins to the site on the plasma membrane, thereby inducing a plurality of recombinant proteins and chimeric proteins to selfassemble into an enveloped nanoparticle (ENP) secreted from the cell in which the recombinant proteins and chimeric proteins are expressed, thereby generating a population of ENPs,optionally the cytoplasmic domain of each of the nth recombinant protein and the cytoplasmic domain of each of the zth chimeric protein are capable of co-localizing each of the nth recombinant protein and each of the zth chimeric protein to a site on the plasma membrane of the cell, optionally the transmembrane domain of each of the nth recombinant protein and the transmembrane domain of each of the zth chimeric protein are capable of co-localizing each of the nth recombinant protein and each of the zth chimeric protein to a site on the plasma membrane of the cell.

33. The composition of any one of claims 31-32, wherein the plurality of recombinant proteins and the plurality of chimeric proteins are capable of being presented on the surface of the cell in which the plurality of recombinant proteins and the plurality of chimeric proteins are expressed, optionally the cell is: a cell of a subject; an in vivo cell, an ex vivo cell, or an in situ cell; and / or an adherent cell or a suspension cell.

34. The composition of any one of claims 31-33, wherein the self-assembly of an ENP: does not require an exogenous nucleic acid other than the nucleic acid composition, and / or does not require any exogenous components other than the recombinant protein and the chimeric protein.

35. The composition of any one of claims 31-34, wherein upon secretion from a cell of a subject, the ENPs are capable of distributing within one or more tissues of the subject, optionally the one or more tissues comprise adrenal gland tissue, appendix tissue, bladder tissue, bone, bowel tissue, brain tissue, breast tissue, bronchi, coronal tissue, ear tissue, esophagus tissue, eye tissue, gall bladder tissue, genital tissue, heart tissue, hypothalamus tissue, kidney tissue, large intestine tissue, intestinal tissue, larynx tissue, liver tissue, lung tissue, lymph nodes, mouth tissue, nose tissue, pancreatic tissue, parathyroid gland tissue, pituitary gland tissue, prostate tissue, rectal tissue, salivary gland tissue, skeletal muscle tissue, skin tissue, small intestine tissue, spinal cord, spleen tissue, stomach tissue, thymus gland tissue, trachea tissue, thyroid tissue, ureter tissue, urethra tissue, soft and connective tissue, peritoneal tissue, blood vessel tissue, fat tissue, or any combination thereof, further optionally the ENPs engage a plurality of immune cells in the one or more tissues, thereby mimicking a natural infection.

36. A composition, comprising: a population of enveloped nanoparticles (ENPs), wherein each of the ENPs comprises a plurality of recombinant proteins comprising a transmembrane domain and an optional antigenic polypeptide (AP), and a plurality of chimeric proteins comprising: a) a signal peptide, b) a cell surface domain, c) a transmembrane domain, and d) a cytoplasmic domain, wherein the plurality of recombinant proteins and / or the plurality of chimeric proteins further comprise an endosomal sorting complex required for transport (ESCRT)-recruiting domain (ERD), optionally the plurality of recombinant proteins further comprise a cytoplasmic domain.

37. The composition of any one of claims 31-36, wherein the ENPs are derived from expression of the nucleic acid composition of any one of claims 31-35.

38. The composition of any one of claims 31-37, wherein the signal peptide comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to an amino acid of SEQ ID NO: 127 or a sequence having one, two, or three mismatches relative to the sequence of SEQ ID NO: 127.

39. The composition of any one of claims 31-38, wherein the cell surface domain comprises an immune-cell targeting polypeptide or a second antigenic polypeptide sequence, optionally the immune-cell targeting polypeptide comprises or is derived from one or more of complement component 3d (C3d), flagellin, an influenza HA molecule, a parainfluenza FiN molecule, a Venezuelan equine encephalitis (VEE) glycoprotein molecule, a mannose receptor molecule, a mammalian toll- like receptor (TLR) ligand molecule, a MIP-1 alpha molecule, a RANTES MIP-1 beta molecule, a GM-CSF molecule, a Flt3 ligand molecule, a CD40 ligand molecule, a Prevotella intermedia glycoprotein, a respiratory syncytial virus protein F, a fibronectin A domain, fibrinogen, a measles virus HA protein, and Pam2Cys lipoprotein / lipopeptide (MALP-2), further optionally wherein the immune-cell targeting polypeptide is capable of targeting the ENP to one or more immune cells each selected from the group comprising: a T-cell, a B-cell, a macrophage, a neutrophil, a dendritic cell, an innate lymphoid cell, a mast cell, an eosinophil, a basophil, a megakaryocyte, or a natural killer cell.

40. The composition of any one of claims 31-39, wherein the cell surface domain and the chimeric protein transmembrane domain are separated by a linker, wherein the linker:is a flexible linker, a rigid linker, or a hybrid linker; is hydrophilic or hydrophobic; is between 1 and 250 amino acids; comprises one or more flexible amino acid residues, optionally about 1 to about 250 flexible amino acid residues, further optionally the flexible amino acid residues comprise glycine, serine, or a combination thereof; and / or comprises 3 repeating amino acid subunits or more.

41. The composition of any one of claims 31-40, wherein: the cell surface domain comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 128 or SEQ ID NO: 130; the cytoplasmic domain and / or transmembrane domain of the recombinant protein comprise an amino acid sequence having at least 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of the cytoplasmic domain and / or the transmembrane domain of the chimeric protein;(i) the transmembrane domain and the cytoplasmic domain of the recombinant protein and (ii) the transmembrane domain and the cytoplasmic of the chimeric protein each comprise an amino acid sequence having at least 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 129; the chimeric protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 124, SEQ ID NO: 125, or SEQ ID NO: 126; and / or the AP of the recombinant protein comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 40 or SEQ ID NO: 47.

42. The composition of any one of claims 31-41, wherein one or more of (i) the polynucleotide encoding the recombinant protein or the n polynucleotides each encoding an nth recombinant protein, and (ii) the polynucleotide encoding the chimeric protein or the z polynucleotides each encoding a zth chimeric protein, are present in a same or different nucleic acid molecule, optionally wherein the amount of(i) the polynucleotide encoding the recombinant protein or the n polynucleotides each encoding an nth recombinant protein; and(ii) the polynucleotide encoding the chimeric protein or the z polynucleotides each encoding a zth chimeric protein,are present in the composition at a molar ratio of about 10: 1, 4: 1, or 1 : 1.

43. The composition of any one of claims 31-42, wherein the polynucleotide encoding the recombinant protein and the polynucleotide encoding the chimeric protein are present in the same nucleic acid molecule.

44. The composition of any one of claims 31-43, wherein the cell expressing the recombinant protein or the plurality of recombinant proteins and the chimeric protein or plurality of chimeric proteins exhibits at least a 2-fold increase in AP expression at the surface of the cell, relative to a cell expressing a fusion protein or a plurality of fusion proteins comprising an AP and an ERD and does not express the chimeric protein.

45. The composition of any one of claims 31-44, wherein the nucleic acid composition further comprises a polynucleotide comprising or encoding a tetherin inhibitor, optionally the tetherin inhibitor is capable of modulating expression, concentration, localization, stability, and / or activity of tetherin and / or wherein presence or expression of the tetherin inhibitor in the cell results in an increase in ENP production by the cell by at least 2-fold, relative to a cell that does not comprise or express the tetherin inhibitor, further optionally the tetherin inhibitor comprises a dsRNA, an siRNA, an shRNA, a pre- miRNA, a pri-miRNA, a miRNA, an stRNA, an IncRNA, a piRNA, a snoRNA, or a protein.

46. The composition of claim 45, wherein one or more of (i) the polynucleotide comprising or encoding the tetherin inhibitor, (ii) the polynucleotide encoding the recombinant protein or the n polynucleotides each encoding an nth recombinant protein, and (iii) the polynucleotide encoding the chimeric protein or the z polynucleotides each encoding a zth chimeric protein, are present in a same or different nucleic acid molecule, optionally wherein the amount of(i) the polynucleotide comprising or encoding the tetherin inhibitor; and(ii) the polynucleotide encoding the recombinant protein or the n polynucleotides each encoding an nth recombinant protein, and / or the polynucleotide encoding the chimeric protein or the z polynucleotides each encoding a zth chimeric protein, are present in the composition at a molar ratio of about 1 : 1, 1 :5 or 1 :25.

47. The composition of claim 45, wherein the polynucleotide comprising or encoding the tetherin inhibitor and the polynucleotide encoding the recombinant protein are present in the same nucleic acid or wherein the polynucleotide comprising or encoding the tetherin inhibitor and the polynucleotide encoding the chimeric protein are present in the same nucleic acid.

48. The composition of any one of claims 45-47, wherein the tetherin inhibitor comprises or is derived from a viral protein, optionally the virus is HIV-1, HIV-2, SIV, Ebolavirus, KSHV, SARS CoV, or SARS-CoV-2, further optionally the tetherin inhibitor comprises HIV-1 Vpu protein, KSHV K5 protein, SARS-CoV-2 ORF7a, HIV-2 Env, Ebola GP, SIV Env, SIV Vpu, SIV Nef, or any portions, variants or derivatives thereof.

49. The composition of any one of claims 45-48, wherein the tetherin inhibitor comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 132-134, optionally the tetherin inhibitor comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the sequence of SEQ ID NO: 132.

50. The composition of any one of claims 1-49, wherein the ENPs comprise a lipid bilayer, optionally a lipid bilayer derived from the cell from which the ENP was secreted.

51. The composition of any one of claims 1-50, wherein the ERD is located at the C- terminus of the dimerization fusion protein, the adapter fusion protein, the recombinant protein and / or the chimeric protein, optionally the ERD is capable of interacting with the ESCRT proteins TSG101, NEDD4, and / or ALIX.

52. The composition of any one of claims 1-51, wherein the ERD comprises or is derived from: a human protein; a nonhuman protein, optionally a nonmammalian protein, further optionally a chicken protein, a mouse protein, a lizard protein, a reptile protein, a hamster protein, or a goldfish protein; the ESCRT and ALIX binding region (EABR) of the human CEP55 protein, optionally residues 170-213;Syntenin-1, rat Galectin-3 (rGalectin-3), Hrs, and / or CD2AP; a viral protein, optionally a fragment of a viral protein, further optionally a retroviral protein, herpes simplex viral protein, vaccinia viral protein, hepadnaviral protein, togaviral protein, flaviviral protein, arenaviral protein, coronaviral protein, orthomyxoviral protein, paramyxoviral protein, bunyaviral protein, bornaviral protein, rhabdoviral protein or filoviral protein, optionally a Gag protein, further optionally derived from EIAV, HTLV- 1, MLV, or MPMV, optionally EIAV p9 and / or HIV-1 p6; and / or an Ebola protein, optionally EBOV VP40, optionally the ERD comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the amino acid sequence of SEQ ID NO: 2.

53. The composition of any one of claims 1-52, wherein the ERD comprises one ormore TSG101 -binding motifs, one or more ALIX-binding motifs, one or more Nedd4-recruiting motifs, or any combination thereof, optionally wherein any two of the one or more TSG101- binding motifs, the one or more ALIX-binding motifs, or the one or more Nedd4-recruiting motifs are the same or different.

54. The composition of any one of claims 1-52, wherein the ERD comprises or is derived from HIV-1 p6 protein or SIV p6 protein, optionally from the p6 protein of HIV-1 isolate ETH2220.

55. The composition of any one of claims 1-52, wherein the ERD comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 5-37.

56. The composition of any one of claims 1-52, wherein the ERD comprises or is derived from a non-human galectin protein, optionally a rat galectin protein, optionally the ERD comprising or derived from a non-human galectin protein further comprises a viral-derived ALIX- recruiting motif, further optionally the ERD comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 38-39.

57. The composition of any one of claims 1-56, wherein the dimerization fusion protein, the adapter fusion protein, the recombinant protein, and / or the chimeric protein comprises an endocytosis-preventing motif (EPM) capable of preventing endocytosis of the dimerization fusion protein, the adapter fusion protein, the recombinant protein, and / or the chimeric protein, optionally the EPM: tethers the dimerization fusion protein, the adapter fusion protein, the recombinant protein, and / or the chimeric protein to the cytoskeleton, thereby preventing localization to coated pits and endocytosis; enhances ENP assembly, ENP production, and / or ENP secretion; and / or prevents endocytosis of the dimerization fusion protein, the adapter fusion protein, the recombinant protein, and / or the chimeric protein, thereby extending the time the dimerization fusion protein, the adapter fusion protein, the recombinant protein, and / or the chimeric protein remains at the plasma membrane to interact with ESCRT proteins, further optionally the EPM: increases the abundance and / or density of dimerization fusion proteins, adapter fusion proteins, recombinant proteins, and / or chimeric proteins on and / or in the ENP by at least about 2-fold as compared to an ENP comprising a dimerization fusion protein, an adapter fusion protein, a recombinant protein, and / or a chimeric protein that does not comprise the EPM; and / orincreases the number of ENPs secreted by a cell by at least about 2-fold as compared to a cell expressing a dimerization fusion protein, an adapter fusion protein, a recombinant protein, and / or a chimeric protein that does not comprise the EPM.

58. The composition of claim 57, wherein the EPM comprises or is derived from a portion of murine low-affinity gamma Fc region receptor II isoform FcRII-Bl, optionally the EPM comprises all or a portion of the cytoplasmic tail of FcRII-Bl, optionally the EPM comprises an amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to the amino acid sequence of SEQ ID NO: 1.

59. The composition of any one of claims 1-56, wherein the dimerization fusion protein, the adapter fusion protein, the recombinant protein, and / or the chimeric protein does not comprise an endocytosis-preventing motif (EPM).

60. The composition of any one of claims 1-59, wherein the AP, or a portion thereof, is displayed in and / or on the surface of the ENP, optionally the AP: is about 1 amino acid to about 10000 amino acids in length; comprises or is derived from an antigenic protein associated with a disease or disorder, optionally an immunogenic variant and / or an immunogenic fragment of said antigenic protein; comprises or is derived from a conserved portion of said antigenic protein; is present on and / or in the ENP in its natural membrane-associated conformation; and / or comprises or is derived from at least about 5 percent of the full length of said antigenic protein, optionally the AP comprises or is derived from the full-length surface protein of an infectious agent.

61. The composition of claim 60, wherein the disease or disorder is an infectious disease or disorder caused by an infectious agent, wherein the AP comprises or is derived from an antigenic protein of said infectious agent, and wherein the antigenic protein of said infectious agent is a pathogenic antigen, optionally wherein the disease or disorder is a disease associated with expression of a tumor-associated antigen, and wherein the antigenic protein is a tumor-associated antigen; wherein the disease or disorder is an autoimmune disease or disorder, and wherein the antigenic protein is an autoimmune antigen; and / or wherein the disease or disorder is an allergic disease or disorder, and wherein the antigenic protein is an allergenic antigen.

62. The composition of any one of claims 60-61, wherein the infectious agent is:a bacterium, a fungus, a virus, or a protist; and / or a coronavirus (CoV), optionally the CoV comprises an alphacoronavirus, a betacoronavirus, a gammacoronavirus, or a deltacoronavirus, optionally wherein the infectious agent is selected from the group comprising Acinetobacter baumannii, Anaplasma genus, Anaplasma phagocytophilum, Ancylostoma braziliense, Ancylostoma duodenale, Arcanobacterium haemolyticum, Ascaris lumbricoides, Aspergillus genus, Astroviridae, Babesia genus, Bacillus anthracis, Bacillus cereus, Bartonella henselae, BK virus, Blastocystis hominis, Blastomyces dermatitidis, Bordetella pertussis, Borrelia burgdorferi, Borrelia genus, Borrelia spp, Brucella genus, Brugia malayi, Bunyaviridae family, Burkholderia cepacia and other Burkholderia species, Burkholderia mallei, Burkholderia pseudomallei, Caliciviridae family, Campylobacter genus, Candida albicans, Candida spp, Chlamydia trachomatis, Chlamydophila pneumoniae, Chlamydophila psittaci, CJD prion, Clonorchis sinensis, Clostridium botulinum, Clostridium difficile, Clostridium perfringens, Clostridium perfringens, Clostridium spp, Clostridium tetani, Coccidioides spp, coronaviruses, Corynebacterium diphtheriae, Coxiella burnetii, Crimean-Congo hemorrhagic fever virus, Cryptococcus neoformans, Cryptosporidium genus, Cytomegalovirus (CMV), Dengue viruses (DEN-1, DEN-2, DEN-3 and DEN-4), Dientamoeba fragilis, Ebolavirus (EBOV), Echinococcus genus, Ehrlichia chaffeensis, Ehrlichia ewingii, Ehrlichia genus, Entamoeba histolytica, Enterococcus genus, Enterovirus genus, Enteroviruses, mainly Coxsackie A virus and Enterovirus 71 (EV71), Epidermophyton spp, Epstein-Barr Virus (EBV), Escherichia coli 0157 :H7 , 0111 and 0104 :H4, Fasciola hepatica and Fasciola gigantica, FFI prion, Filarioidea superfamily, Filoviruses, Flaviviruses, Francisella tularensis, Fusobacterium genus, Geotrichum candidum, Giardia intestinalis, Gnathostoma spp, GSS prion, Guanarito virus, Haemophilus ducreyi, Haemophilus influenzae, Helicobacter pylori, Henipavirus (Hendra virus Nipah virus), Hepatitis A Virus, Hepatitis B Virus (HBV), Hepatitis C Virus (HCV), Hepatitis D Virus, Hepatitis E Virus, Herpes simplex virus 1 and 2 (HSV-1 and HSV-2), Histoplasma capsulatum, HIV (Human immunodeficiency virus), Hortaea werneckii, Human bocavirus (HBoV), Human herpesvirus 6 (HHV-6) and Human herpesvirus 7 (HHV-7), Human metapneumovirus (hMPV), Human papillomavirus (HPV), Human parainfluenza viruses (HPIV), Japanese encephalitis virus, JC virus, Junin virus, Kingella kingae, Klebsiella granulomatis, Kuru prion, Lassa virus, Legionella pneumophila,Leishmania genus, Leptospira genus, Listeria monocytogenes, Lymphocyticchoriomeningitis virus (LCMV), Machupo virus, Malassezia spp, Marburg virus, Measles virus, Metagonimus yokagawai, Microsporidia phylum, Molluscum contagiosum virus (MCV), Mumps virus, Mycobacterium leprae and Mycobacterium lepromatosis, Mycobacterium tuberculosis, Mycobacterium ulcerans, Mycoplasma pneumoniae, Naegleria fowleri, Necator americanus, Neisseria gonorrhoeae, Neisseria meningitidis, Nocardia asteroides, Nocardia spp, Onchocerca volvulus, Orientia tsutsugamushi, Orthomyxoviridae family (Influenza), Paracoccidioides brasiliensis, Paragonimus spp, Paragonimus westermani, Parvovirus Bl 9, Pasteurella genus, Plasmodium genus, Pneumocystis jirovecii, Poliovirus, Rabies virus, Respiratory syncytial virus (RSV), Rhinovirus, rhinoviruses, Rickettsia akari, Rickettsia genus, Rickettsia prowazekii, Rickettsia rickettsii, Rickettsia typhi, Rift Valley fever virus, Rotavirus, Rubella virus, Sabia virus, Salmonella genus, Sarcoptes scabiei, SARS coronavirus, Schistosoma genus, Shigella genus, Sin Nombre virus, Hantavirus, Sporothrix schenckii, Staphylococcus genus, Staphylococcus genus, Streptococcus agalactiae, Streptococcus pneumoniae, Streptococcus pyogenes, Strongyloides stercoralis, Taenia genus, Taenia solium, Tick-borne encephalitis virus (TBEV), Toxocara canis or Toxocara cati, Toxoplasma gondii, Treponema pallidum, Trichinella spiralis, Trichomonas vaginalis, Trichophyton spp, Trichuris trichiura, Trypanosoma brucei, Trypanosoma cruzi, Ureaplasma urealyticum, Varicella zoster virus (yZN Variola major or Variola minor, vCJD prion, Venezuelan equine encephalitis virus, Vibrio cholerae, West Nile virus, Western equine encephalitis virus, Wuchereria bancrofti, Yellow fever virus, Yersinia enterocolitica, Yersinia pestis, and Yersinia pseudotuberculosis.

63. The composition of any one of claims 1-62, wherein the AP: comprises a membrane protein, optionally a multi-span transmembrane protein, and / or is not configured to be a soluble protein, optionally the AP does not comprise one or more mutations configured to enhance its solubility and / or stability, or wherein the AP does not comprise a transmembrane domain and / or is a soluble protein, and wherein the dimerization fusion protein and the adapter fusion protein comprises a transmembrane domain (TD), optionally the transmembrane domain comprises or is derived from a human transmembrane protein or a nonhuman transmembrane protein, optionally a nonmammalian transmembrane protein, further optionally the TD comprises or is derived from a natural protein, a recombinant protein, and / or synthetic protein, optionally a synthetic protein comprising predominantly hydrophobic residues.

64. The composition of any one of claims 1-63, wherein the population of ENPs comprise: one or more homotypic ENPs, wherein the plurality of dimerization fusion proteins or the plurality of recombinant proteins of a homotypic ENP are the same as each other with respect to the AP, and wherein a homotypic ENP thereby does not display a plurality of disparate AP; one or more heterotypic ENPs, wherein at least two of the plurality of dimerization fusion proteins or the plurality of recombinant proteins of a heterotypic ENP are different from each other with respect to the AP, and wherein a heterotypic ENP thereby displays a plurality of disparate AP; a mixture of two or more homotypic ENPs that differ from each other with respect to the AP of the plurality of dimerization fusion proteins or the plurality of recombinant proteins present in said two or more homotypic ENPs, and wherein the population of ENPs thereby displays a plurality of disparate AP; and / or a mixture of two or more heterotypic ENPs that differ from each other with respect to the AP of the plurality of dimerization fusion proteins or the plurality of recombinant proteins of said two or more heterotypic ENPs, optionally heterotypic ENPs are capable of eliciting heterologous antibody responses against an additional infectious agent, and wherein said heterotypic ENPs do not display AP derived from said additional infectious agent.

65. The composition of claim 64, wherein the plurality of disparate AP comprises: between about 2 and about 500 antigenic polypeptides that differ from each other;AP of a same protein type; and / orAP of different protein types, optionally the same ENP comprises the plurality of disparate AP derived from two or more strains of the same family, same genus, and / or same species, of infectious agent, further optionally the plurality of disparate AP has a sequence identity of about, at least, or at least about 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% with one another.

66. The composition of any one of claims 64-65, wherein the plurality of disparate AP comprise a plurality of coronavirus (CoV) antigens, wherein the plurality of CoV antigens comprises a first CoV antigen of a first CoV and a second CoV antigen of a second CoV that is different from the first CoV, optionally the plurality of CoV antigens comprise a CoV spike protein (S protein) or a portion thereof, a CoV envelope protein (E protein) or a portion thereof, a CoV nucleocapsidprotein (N protein) or a portion thereof, a CoV hemagglutinin-esterase protein (HE protein) or a portion thereof, a CoV papain-like protease or a portion thereof, a CoV 3 CL protease or a portion thereof, a CoV membrane protein (M protein) or a portion thereof, or a combination thereof.

67. The composition of claim 66, wherein: the first CoV antigen, the second CoV antigen, or both comprise a CoV S protein or a portion thereof; the number of the first CoV antigen molecules and the number of the second CoV antigen molecules are in a ratio from 1 : 100 to 100: 1; the plurality of CoV antigens comprise three, four, five, size seven, or eight CoV antigens, each of a CoV different from one another; and / or the plurality of CoV antigens further comprise at least a third CoV antigen of a third CoV and a fourth CoV antigen of a fourth CoV, and wherein the first, second, third and fourth CoVs are different from one another.

68. The composition of any one of claims 64-67, wherein the plurality of disparate AP comprise at least m pathogenic antigens of an mth infectious agent, wherein m is an integer greater than 2, and wherein each mth pathogenic antigen is different from one another, optionally m is an integer greater than 50; and / or wherein the plurality of disparate AP comprise one or more of a 1st pathogenic antigen (PA) of a 1st infectious agent (IA), a 2nd PA of a 2nd IA, a 3rd PA of a 3rd IA, a 4th PA of a 4th IA, a 5th PA of a 5th IA, a 6th PA of a 6th IA, a 7th PA of a 7th IA, a 8th PA of a 8th IA, a 9th PA of a 9th IA, a 10th PA of a 10th IA, a 11th PA of a 11th IA, a 12th PA of a 12th IA, a 13th PA of a 13th IA, a Mth PA of a Mth IA, a 15th PA of a 15th IA, a 16th PA of a 16th IA, a 17th PA of a 17th IA, a 18th PA of a 18th IA, a 19th PA of a 19th IA, a 20th PA of a 20th IA, a 21 st PA of a 21 st IA, a 22nd PA of a 22nd IA, a 23rd PA of a 23rd IA, a 24th PA of a 24th IA, a 25th PA of a 25th IA, a 26th PA of a 26th IA, a 27th PA of a 27th IA, a 28th PA of a 28th IA, a 29th PA of a 29th IA, a 30th PA of a 30th IA, a 31st PA of a 31st IA, a 32nd PA of a 32nd IA, a 33rd PA of a 33rd IA, a 34th PA of a 34th IA, a 35th PA of a 35th IA, a 36th PA of a 36th IA, a 37th PA of a 37th IA, a 38th PA of a 38th IA, a 39th PA of a 39th IA, a 40th PA of a 40th IA, a 41st PA of a 41st IA, a 42nd PA of a 42nd IA, a 43rd PA of a 43rd IA, a 44th PA of a 44th IA, a 45th PA of a 45th IA, a 46th PA of a 46th IA, a 47th PA of a 47th IA, a 48th PA of a 48th IA, a 49th PA of a 49th IA, and a 50th PA of a 50th IA, wherein the 1st, 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, 9th, 10th, 11th, 12th, 13th, Mth, 15th, 16th, 17th, 18th, 19th, 20th, 21st, 22nd, 23rd, 24th, 25th, 26th, 27th, 28th, 29th, 30th, 31st, 32nd, 33rd, 34th, 35th, 36th, 37th, 38th, 39th,40th, 41st, 42nd, 43rd, 44th, 45th, 46th, 47th, 48th, 49th, and 50th pathogenic antigens are different from one another, further optionally the 1st, 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, 9th, 10th, 11th, 12th, 13th, 14th, 15th, 16th, 17th, 18th, 19th, 20th, 21st, 22nd, 23rd, 24th, 25th, 26th, 27th, 28th, 29th, 30th, 31st, 32nd, 33rd, 34th, 35th, 36th, 37th, 38th, 39th, 40th, 41st, 42nd, 43rd, 44th, 45th, 46th, 47th, 48th, 49th, and 50th infectious agents are different from one another, optionally the plurality of disparate AP comprise the plurality of CoV antigens, a plurality of influenza antigens, and / or a plurality of HIV antigens, optionally one or more of the plurality of CoV antigens are of CoVs in the genus of Alpha-CoV and / or Beta-CoV, and optionally wherein each of the plurality of CoV antigens are of Co Vs in the genus of Beta-CoV.

69. The composition of any one of claims 66-68, wherein: the plurality of CoV antigens are of Co Vs in the subgenus of Sarbecovirus; the first CoV and the second CoV are in the genus of Beta-CoV, optionally in the subgenus of Sarbecovirus; the plurality of CoV antigens are of CoVs selected from the group consisting of: SARS-CoV, SARS-CoV-2, WIV1, SHC014, Rfl, RmYN02, pangl7, RaTG13, Rs4081, LYRal l, HKU3, Yunnan2011, BtKY72, BM48-31, WIV16, Khosta-1, and Khosta-2; the first CoV, the second CoV, or both are selected from the group consisting of: SARS-CoV, SARS-CoV-2, WIV1, SHC014, Rfl, RmYN02, pangl7, RaTG13, Rs4081, LYRal l, HKU3, Yunnan2011, BtKY72, BM48-31, WIV16, Khosta-1, and Khosta-2; the CoV is selected from a species or subspecies of SARS-CoV, SARS-CoV- 1, SARS-CoV-2, MERS-CoV, SL-CoV-WIVl, HKU4, HKU5, HCoV-OC43, HCoV-HKUl, HKU9, HKU3, HKU8, HKU24, NL63, SHC014, 229E and / or SARS-CoV-2 variants B.1.351, B.1.1.7, P.l, B.1.617.2, B.1.1.529, BA. l, BA.1.1, BA.2, BA.3, BA.4, BA.5 and other descendent lineages; the CoV is selected from a species or subspecies of Embecovirus, Sarbecovirus, Merbecovirus, Nobevovirus, Hibecovirus, SARSr-CoV, MERS-CoV, or any combination thereof; and / or the CoV is selected from a beta-CoV from the sarbe-, embeco-, merbeco-, and nobecovirus lineages.

70. The composition of any one of claims 66-69, wherein the CoV is selected from a a sarbecovirus strain, optionally, SARS, LYRal l, Rfl, Rs4081, BtKY72, and / orBM48-31; a merbecovirus strain, optionally HKU4, HKU5, HKU25, BtCoV-Vs-CoVl, MERS-related NL13845, and MERS-related NL 140422; and / oran embecovirus strain, optionally HKU1, Rat CoV Parker, PHEV, Equine CoV, Rodent CoV, and Longquan Rat CoV.

71. The composition of any one of claims 1-70, wherein: the ENPs comprise at least about 2-fold more of the AP and / or are at least as immunogenic as compared to a multi-component nanoparticle approach, optionally as compared to a SpyCatcher-based nanoparticle approach or a lentiviral Gag-based approach, further optionally the multi-component nanoparticle approach comprises two or more separate polypeptides; and / or the ENPs comprise at least about 2-fold more of the AP, an at least about 2-fold higher density of the AP, and / or are at least as immunogenic, as compared to a nanoparticle approach that does not comprise the ERD, optionally as compared to a SpyCatcher-based or Gag-based nanoparticle approach, optionally the nucleic acid composition does not comprise a polynucleotide encoding SpyTag or lentiviral Gag; and / or the ENPs do not comprise SpyTag or lentiviral Gag.

72. The composition of any one of claims 1-71, wherein: the ENPs have one or more dimensions of a eukaryotic virus; less than about 10% of the ENPs of the population of ENPs have a particle size smaller than about 10 nm; less than about 10% of the ENPs of the population of ENPs have a particle size exceeding about 80 nm; the average diameter of the ENPs of the population of ENPs range from about 5 nm to about 80 nm, from about 15 nm to about 50 nm, or from about 20 nm to about 40 nm; and / or the average diameter of the ENPs of the population of ENPs is about 10 nm, about 12 nm, about 14 nm, about 16 nm, about 18 nm, about 20 nm, about 22 nm, about 24 nm, about 26 nm, about 28 nm, about 30 nm, about 32 nm, about 34 nm, about 36 nm, about 38 nm, about 40 nm, about 42 nm, about 44 nm, about 46 nm, about 48 nm, or about 50 nm, optionally the average is the mean, median or mode, optionally the mean is the arithmetic mean, geometric mean, and / or harmonic mean, optionally the ENPs: have a minimum diameter of about 10 nm, about 12 nm, about 14 nm, about 16 nm, about 18 nm, about 20 nm, about 22 nm, about 24 nm, about 26 nm, about 28 nm, about 30 nm, about 32 nm, about 34 nm, about 36 nm, about 38 nm, about 40 nm, about 42 nm, about 44 nm, about 46 nm, about 48 nm, or about 50 nm;have a maximum diameter of about 10 nm, about 12 nm, about 14 nm, about 16 nm, about 18 nm, about 20 nm, about 22 nm, about 24 nm, about 26 nm, about 28 nm, about 30 nm, about 32 nm, about 34 nm, about 36 nm, about 38 nm, about 40 nm, about 42 nm, about 44 nm, about 46 nm, about 48 nm, about 50 nm, about 52 nm, about 54 nm, about 56 nm, about 58 nm, about 60 nm, about 62 nm, about 64 nm, about 66 nm, about 68 nm, about 70 nm, about 72 nm, about 74 nm, about 76 nm, about 78 nm, or about 80 nm; and / or are derived from cell cultures transiently transfected with the nucleic acid composition, optionally derived via ultracentrifugation and / or size exclusion chromatography, further optionally ultracentrifugation on a 20% sucrose cushion, optionally transfected via calcium phosphate transfection, DEAE-dextran mediated transfection, cationic lipid-mediated transfection, electroporation, electrical nuclear transport, chemical transduction, electrotransduction, Lipofectamine-mediated transfection, Effectene-mediated transfection, lipid nanoparticle (LNP)-mediated transfection, or any combination thereof.

73. The composition of any one of claims 1-72, wherein: storage of the ENPs at 4°C for at least three months reduces immunogenicity less than about 50 percent; the composition is stable for at least about 2 weeks, about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, or about 1 year, after storage as a liquid at a temperature of about 4°C; and / or at least about 70%, 75%, 80%, 85%, 90% or 95% of the ENPs are immunogenic at least 1 month after storage as a liquid at a temperature of about 5°C, optionally the nucleic acid composition is complexed or associated with one or more lipids or lipid-based carriers, thereby forming liposomes, lipid nanoparticles (LNPs), lipoplexes, and / or nanoliposomes, optionally encapsulating the nucleic acid composition.

74. The composition of any one of claims 1-73, wherein the nucleic acid composition is, comprises, or further comprises, one or more vectors, optionally at least one of the one or more vectors is a viral vector, a plasmid, a transposable element, a naked DNA vector, a lipid nanoparticle (LNP), or any combination thereof, optionally the viral vector is an AAV vector, a lentivirus vector, a retrovirus vector, an adenovirus vector, a herpesvirus vector, a herpes simplex virus vector, a cytomegalovirus vector, a vaccinia virus vector, a MVA vector, a baculovirus vector, a vesicular stomatitis virus vector, a human papillomavirus vector, an avipox virus vector, aSindbis virus vector, a VEE vector, a Measles virus vector, an influenza virus vector, a hepatitis B virus vector, an integration-deficient lentivirus (IDLV) vector, or any combination thereof, and optionally the transposable element is piggybac transposon or sleeping beauty transposon.

75. The composition of any one of claims 1-74, wherein the polynucleotide(s) encoding the dimerization fusion protein, the adapter fusion protein, the recombinant protein, and / or the chimeric protein are comprised in the one or more vectors, optionally the polynucleotide(s) encoding any of the dimerization fusion protein, the adapter fusion protein, the recombinant protein, and / or the chimeric protein are comprised in the same vector and / or different vectors, optionally the polynucleotide(s) encoding any of the dimerization fusion protein, the adapter fusion protein, the recombinant protein, and / or the chimeric protein are situated on the same nucleic acid and / or different nucleic acids.

76. The composition of any one of claims 1-75, wherein the one or more vectors is a DNA vaccine, optionally the polynucleotide(s) encoding any of the dimerization fusion protein, the adapter fusion protein, the recombinant protein, and / or the chimeric protein are operably linked to one or more promoters capable of inducing transcription of said polynucleotide(s), optionally the DNA vaccine is a plasmid-based DNA vaccine, a minicircle-based DNA vaccine, a bacmid-based DNA vaccine, a minigene-based DNA vaccine, a ministring DNA (linear covalently closed DNA vector) vaccine, a closed-ended linear duplex DNA (CELiD or ceDNA) vaccine, a doggybone™ DNA vaccine, a dumbbell shaped DNA vaccine, or a minimalistic immunological-defined gene expression (MIDGE)-vector DNA vaccine, optionally the DNA vaccine elicits at least 2-fold higher neutralizing antibody responses against an infectious agent as compared to a DNA vaccine that encodes the AP only, further optionally wherein: the promoter comprises a ubiquitous promoter, an inducible promoter, a tissuespecific promoter and / or a lineage-specific promoter, optionally the ubiquitous promoter is selected from the group comprising a cytomegalovirus (CMV) immediate early promoter, a CMV promoter, a viral simian virus 40 (SV40) (e.g., early or late), a Moloney murine leukemia virus (MoMLV) LTR promoter, a Rous sarcoma virus (RSV) LTR, anRSV promoter, a herpes simplex virus (HSV) (thymidine kinase) promoter, H5, P7.5, and Pl l promoters from vaccinia virus, an elongation factor 1 -alpha (EFla) promoter, early growth response 1 (EGR1), ferritin H (FerH), ferritin L (FerL), Glyceraldehyde 3- phosphate dehydrogenase (GAPDH), eukaryotic translation initiation factor 4A1 (EIF4A1), heat shock 70 kDa protein 5 (HSPA5), heat shock protein 90 kDa beta, member 1 (HSP90B1), heat shock protein 70 kDa (HSP70), P-kinesin (P-KIN), the human ROSA 26 locus, a Ubiquitin C promoter (UBC), a phosphoglycerate kinase- 1 (PGK) promoter, 3 -phosphoglycerate kinase promoter, a cytomegalovirus enhancer, human P-actin (HBA) promoter, chicken P-actin (CBA) promoter, a CAG promoter, a CASI promoter, a CBH promoter, or any combination thereof; the polynucleotide(s) encoding any of the dimerization fusion protein, the adapter fusion protein, the recombinant protein, and / or the chimeric protein are operably linked to a tandem gene expression element, optionally the tandem gene expression element is an internal ribosomal entry site (IRES), foot-and-mouth disease virus 2A peptide (F2A), equine rhinitis A virus 2A peptide (E2A), porcine teschovirus 2A peptide (P2A) or Thosea asigna virus 2A peptide (T2A), or any combination thereof; and / or the polynucleotide(s) encoding any of the dimerization fusion protein, the adapter fusion protein, the recombinant protein, and / or the chimeric protein comprises a transcript stabilization element, optionally the transcript stabilization element comprises woodchuck hepatitis post-translational regulatory element (WPRE), bovine growth hormone polyadenylation (bGH-polyA) signal sequence, human growth hormone polyadenylation (hGH-polyA) signal sequence, or any combination thereof.

77. The composition of any one of claims 1-76, wherein the nucleic acid composition is or comprises mRNA, optionally the mRNA is formulated in a lipid nanoparticle (LNP), further optionally wherein the wt / wt ratio of lipid to mRNA is from about 1 : 100 to about 100: 1.

78. The composition of claim 77, wherein the mRNA comprises: a 5' untranslated region (UTR), a 3' UTR, and / or a cap; one or more modified nucleotides selected from the group comprising pseudouridine, N-l-methyl-pseudouridine, 2-aminoadenosine, 2-thiothymidine, inosine, pyrrolo-pyrimidine, 3-methyl adenosine, 5-methylcytidine, C-5 propynyl-cytidine, C-5 propynyl-uridine, 2-aminoadenosine, C5-bromouridine, C5-fluorouridine, C5- iodouridine, C5-propynyl-uridine, C5-propynyl-cytidine, C5-methylcytidine, 2- aminoadenosine, 7-deazaadenosine, 7-deazaguanosine, 8-oxoadenosine, 8-oxoguanosine, 0(6)-methylguanine, and 2-thiocytidine; and / or a modified nucleotide in place of one or more uridines, optionally the modifiednucleoside is selected from pseudouridine (y), N 1-methyl-pseudouridine (m IT), and 5- methyl-uridine (m5U).

79. The composition of any one of claims 77-78, wherein the LNP comprises: one or more of an ionizable cationic lipid, a non-cationic lipid, a sterol, and a PEG- modified lipid, optionally the non-cationic lipid is a neutral lipid;0.5-15 mol% PEG-modified lipid, 5-25 mol% non-cationic lipid, 25-55 mol% sterol, and 20-60 mol% ionizable cationic lipid; and / or40-55 mol% ionizable cationic lipid, 5-15 mol% neutral lipid, 35-45 mol% sterol, and 1-5 mol% PEG-modified lipid.

80. The composition of any one of claims 77-79, wherein the LNP comprises:47 mol% ionizable cationic lipid, 11.5 mol% neutral lipid, 38.5 mol% sterol, and 3.0 mol% PEG-modified lipid;48 mol% ionizable cationic lipid, 11 mol% neutral lipid, 38.5 mol% sterol, and 2.5 mol% PEG-modified lipid;49 mol% ionizable cationic lipid, 10.5 mol% neutral lipid, 38.5 mol% sterol, and 2.0 mol% PEG-modified lipid;50 mol% ionizable cationic lipid, 10 mol% neutral lipid, 38.5 mol% sterol, and 1.5 mol% PEG-modified lipid; or51 mol% ionizable cationic lipid, 9.5 mol% neutral lipid, 38.5 mol% sterol, and 1.0 mol% PEG-modified lipid, optionally the ionizable cationic lipid is heptadecan-9-yl 8 ((2 hydroxy ethyl)(6 oxo 6-(undecyloxy)hexyl)amino)octanoate; the neutral lipid is 1,2 distearoyl-sn-glycero-3 phosphocholine (DSPC); the sterol is cholesterol; and / or the PEG-modified lipid is l-monomethoxypolyethyleneglycol-2,3- dimyristylglycerol with polyethylene glycol of average molecular weight 2000 (PEG2000 DMG).

81. The composition of any one of claims 1-80, wherein (i) the nucleic acid composition comprises one or more polynucleotides encoding immunostimulatory agents; and / or (ii) the population of ENPs comprises one or more immunostimulatory agents, optionally the immunostimulatory agents are selected from the group comprising toll-like receptor (TLR) agonists, cytokine receptor agonists, CD40 agonists, Fc receptor agonists, CpG-containing nucleic acids, complement receptor agonists, or any combination thereof, further optionally:the TLR agonist is a TLR-1 agonist, TLR-2 agonist, TLR-3 agonist, TLR-4 agonist, TLR-5 agonist, TLR-6 agonist, TLR-7 agonist, TLR-8 agonist, TLR-9 agonist, and / or TLR- 10 agonist; the Fc receptor agonist is a Fc-gamma receptor agonist; the complement receptor agonist binds to CD21 or CD35; the complement receptor agonist induces endogenous complement opsonization of the ENP; the cytokine receptor agonist is a cytokine; and / or the cytokine receptor agonist is a small molecule, antibody, fusion protein, or aptamer.

82. The composition of any one of claims 1-81, wherein the composition: further comprises Tris buffer, sucrose, and / or sodium acetate; further comprises an adjuvant, optionally the adjuvant is selected from the group comprising aluminum hydroxide, alhydrogel, AddaVax, MF59, AS03, Freund’s adjuvant, Montanide ISA51, CpG, Poly I:C, glucopyranosyl lipid A, flagellin, resiquimod, or any combination thereof; is a lyophilized composition, optionally the lyophilized composition has a water content of less than about 10%; and / or is formulated or is to be formulated: as a liquid, a solid, or a combination thereof; for injection; for intramuscular administration, intranasal administration, transdermal administration, aerosol delivery, nasal delivery, vaginal delivery, rectal delivery, buccal delivery, ocular delivery, local delivery, topical delivery, intracistemal delivery, intraperitoneal delivery, oral delivery, intramuscular injection, intravenous injection, subcutaneous injection, intranodal injection, intratumoral injection, intraperitoneal injection, intradermal injection; and / or as particles, optionally the particles are iron oxide particles, liposomes, micelles, polymer complexes, cationic peptide nanoemulsions, virus-like particles (VLPs), lipid nanoparticles (LNP) and / or lipoplex (LPX) particles, optionally the nucleic acid composition and the LNP-forming components are in separate vials.

83. The composition of any one of claims 1-82, wherein the composition is a pharmaceutical composition, wherein the pharmaceutical composition further comprises one or more pharmaceutically acceptable carriers, diluents and / or excipients,optionally the composition further comprises instructions for use of the composition for: stimulating an immune response in a subject in need thereof; treating or preventing a disease or disorder caused by an infectious agent in a subject in need thereof; and / or treating or preventing a CoV infection in a subject in need thereof.

84. A kit, comprising the composition of any one of claims 1-83.

85. A cell comprising the nucleic acid composition of any one of claims 1-83.

86. A method of stimulating an immune response in a subject in need thereof, comprising: administering to the subject a pharmaceutically effective amount of the composition of any one of claims 1-83, thereby stimulating an immune response in the subject.

87. A method of treating or preventing a disease or disorder in a subj ect in need thereof, comprising: administering to the subject a pharmaceutically effective amount of the composition of any one of claims 1-83, thereby treating or preventing the disease or disorder in the subject, optionally the disease or disorder is a disease or disorder caused by an infectious agent.

88. The method of claim 87, wherein the disease or disorder caused by an infectious agent is a disease or disorder caused by a coronavirus (CoV) infection.

89. A method for treating or preventing a coronavirus (CoV) infection in a subject in need thereof, comprising: administering to the subject a pharmaceutically effective amount of the composition of any one of claims 1-83, thereby treating or preventing the CoV infection in the subj ect.

90. The method of any one of claims 86-89, wherein: immunogenic levels of any of the dimerization fusion protein, the adapter fusion protein, the recombinant protein, and / or the chimeric protein, and / or ENP are produced in serum of the subject at about 1 hour to about 6 months post administration of the composition; a neutralizing antibody titer of about 50 to about 100000 half-maximal inhibitory dilutions (ID50s values) is produced in the serum of the subject at about 1 hour to about 6 months post administration of the composition; and / or the composition elicits at least about 2-fold less off-target immune responses againstundesired epitopes as compared to a non-enveloped NP-based composition, optionally said undesired epitopes comprise the NP scaffold of said non-enveloped NP-based composition.

91. The method of any one of claims 86-90, wherein the method comprises administering to the subject at least two doses of the composition, optionally a second dose of the composition is administered to the subject at least 14 days after a first dose of the composition is administered to the subject.

92. The method of any one of claims 86-91, wherein administering the composition induces neutralizing responses against: the infectious agent(s) from which the antigenic polypeptide(s) are derived; additional infectious agent(s) from which the antigenic polypeptide(s) are not derived, optionally different from the 1st, 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, 9th, 10th, 11th, 12th, 13th, 14th, 15th, 16th, 17th, 18th, 19th, 20th, 21st, 22nd, 23rd, 24th, 25th, 26th, 27th, 28th, 29th, 30th, 31st, 32nd, 33rd, 34th, 35th, 36th, 37th, 38th, 39th, 40th, 41st, 42nd, 43rd, 44th, 45th, 46th, 47th, 48th, 49th, and / or 50th infectious agent; the coronaviruses the plurality of coronavirus antigens are of; coronaviruses different from the 1st, 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, 9th, 10th, 11th, 12th, 13th, 14th, 15th, 16th, 17th, 18th, 19th, 20th, 21st, 22nd, 23rd, 24th, 25th, 26th, 27th, 28th, 29th, 30th, 31st, 32nd, 33rd, 34th, 35th, 36th, 37th, 38th, 39th, 40th, 41st, 42nd, 43rd, 44th, 45th, 46th, 47th, 48th, 49th, and / or 50th CoV; and / or additional coronaviruses different from the coronaviruses the plurality of coronavirus antigens are of.

93. The method of any one of claims 86-92, wherein administering the composition results in treating or preventing: infection caused by a coronavirus different from the first coronavirus and the second coronavirus; infection caused by additional coronaviruses different from the coronaviruses the plurality of coronavirus antigens are of; infection caused by the coronaviruses the plurality of coronavirus antigens are of; the disease or disorder caused by a coronavirus different from the first coronavirus and the second coronavirus; the disease or disorder caused by additional coronaviruses different from the coronaviruses the plurality of coronavirus antigens are of; and / or the disease or disorder caused by the coronaviruses the plurality of coronavirus antigens are of.

94. The method of any one of claims 86-93, wherein:the composition elicits an at least 2-fold higher neutralizing antibody titer as compared to an approach comprising administration of (i) a soluble version of the AP, and / or (ii) an mRNA vaccine encoding the AP and not encoding the ERD; the composition elicits an at least as high neutralizing antibody titer as compared to an approach comprising administration of a protein-based nanoparticle presenting the AP, optionally administration of the composition elicits protective and long- lasting immunity against the infectious agent(s) and variants thereof; an at least as low dose of the composition is needed to generate a comparable immune response as compared to an approach comprising administration of a proteinbased nanoparticle presenting the AP; and / or an at least about 2-fold lower dose of the composition is needed to generate a comparable immune response as compared to an approach comprising administration of (i) a soluble version of the AP, and / or (ii) an mRNA vaccine encoding the AP and not encoding the ERD.

95. The method of any one of claims 86-94, wherein following administration of the first dose or the second dose of the composition, the composition induces: an at least as potent serum neutralizing titers against the infectious agent or variants thereof as compared to an approach comprising administration of a protein-based nanoparticle presenting the AP, optionally the composition comprises a population of ENPs; an at least 2-fold more potent serum neutralizing titers against the infectious agent or variants thereof as compared to an approach comprising administration of a soluble version of the AP, optionally the composition comprises a population of ENPs; and / or an at least 2-fold more potent serum neutralizing titers against the infectious agent or variants thereof as compared to an approach comprising administration of an mRNA vaccine encoding the AP and not encoding the ERD, optionally the composition comprises an mRNA vaccine encoding a dimerization fusion protein or recombinant protein comprising the AP, optionally as measured by geometric means for serum half-maximal inhibitory dilutions (ID50s values) against the infectious agent or variants thereof, further optionally about 1 day to about 6 months after administration of the first dose of the composition or about 1 day to about 6 months after administration of the second dose of the composition.

96. The method of any one of claims 86-95, wherein the subject: is a human subject;is a newborn or infant of an age of not more than 3 years, of not more than 2 years, of not more than 1.5 years, of not more than 1 year (12 months), of not more than 9 months, 6 months or 3 months, or is between 6 months and 2 years; is immunocompromised, has a pulmonary disease, and / or is 65 years of age or older; has a chronic pulmonary disease, optionally chronic obstructive pulmonary disease (COPD) or asthma; and / or has an underlying comorbid condition, optionally selected from heart disease, diabetes, and lung disease.

97. The method of any one of claims 86-96, wherein the composition is administered in an effective amount to: induce a robust antibody response against the AP in the subject, optionally a robust antibody response comprises a neutralizing antibody response, further optionally a robust antibody response comprises Fc domain effector functions that recruit immune cells to infected cells, optionally said immune cells are macrophages, neutrophils, and / or natural killer cells, further optionally said recruitment induces antibody-dependent cellular cytotoxicity (ADCC) and / or antibody-dependent cellular phagocytosis (ADCP); elicit a robust CD4 and / or CD8 T cell response against the AP in the subject; and / or elicit a balanced Thl / Th2 response against the AP in the subject.

98. The method of any one of claims 86-97, wherein the composition is: (i) coadministered with an adjuvant; or (ii) not co-administered with an adjuvant, optionally wherein administering comprises aerosol delivery, nasal delivery, vaginal delivery, rectal delivery, buccal delivery, ocular delivery, local delivery, topical delivery, intracisternal delivery, intraperitoneal delivery, oral delivery, intramuscular injection, intravenous injection, subcutaneous injection, intranodal injection, intratumoral injection, intraperitoneal injection, intradermal injection, or any combination thereof, optionally the composition is administered intramuscularly, further optionally into a deltoid region of an arm.

99. The method of any one of claims 86-98, wherein: the disease or disorder is a blood disease, an immune disease, a neurological disease or disorder, a cancer, an infectious disease, a genetic disease, a disorder caused by aberrant mtDNA, a metabolic disease, a disorder caused by aberrant cell cycle, a disorder caused by aberrant angiogenesis, a disorder cause by aberrant DNA damage repair, or any combination thereof, optionally a solid tumor; the disease or disorder is an infectious disease selected from the group consisting of an Acute Flaccid Myelitis (AFM), Anaplasmosis, Anthrax, Babesiosis, Botulism,Brucellosis, Campylobacteriosis, Carbapenem-resistant Infection, Chancroid, Chikungunya Virus Infection, Chlamydia, Ciguatera, Difficile Infection, Perfringens, Coccidioidomycosis fungal infection, coronavirus infection, Covid-19 (SARS-CoV-2), Creutzfeldt-Jacob Disease / transmissible spongiform encephalopathy, Cryptosporidiosis (Crypto), Cyclosporiasis, Dengue 1,2,3 or 4, Diphtheria, E. coli infection / Shiga toxinproducing (STEC), Eastern Equine Encephalitis, Hemorrhagic Fever (Ebola), Ehrlichiosis, Encephalitis, Arboviral or parainfectious, Non-Polio Enterovirus, D68 Enter oviru(EV- D68), Giardiasis, Glanders, Gonococcal Infection, Granuloma inguinale, Haemophilus Influenza disease Type B (Hib or H-flu), Hantavirus Pulmonary Syndrome (HPS), Hemolytic Uremic Syndrome (HUS), Hepatitis A (Hep A), Hepatitis B (Hep B), Hepatitis C (Hep C), Hepatitis D (Hep D), Hepatitis E (Hep E), Herpes, Herpes Zoster (Shingles), Histoplasmosis infection, Human Immunodeficiency Virus / AIDS (HIV / AIDS), Human Papillomavirus (HPV), Influenza (Flu), Legionellosis (Legionnaires Disease), Leprosy (Hansens Disease), Leptospirosis, Listeriosis (Listeria), Lyme Disease, Lymphogranuloma venereum infection (LGV), Malaria, Measles, Melioidosis, Meningitis (Viral), Meningococcal Disease (Meningitis (Bacterial)), Middle East Respiratory Syndrome Coronavirus (MERS-CoV), Mumps, Norovirus, Pediculosis, Pelvic Inflammatory Disease (PID), Pertussis (Whooping Cough), Plague (Bubonic, Septicemic, Pneumonic), Pneumococcal Disease (Pneumonia), Poliomyelitis (Polio), Powassan, Psittacosis, Pthiriasis, Pustular Rash diseases (Small pox, monkeypox, cowpox), Q-Fever, Rabies, Rickettsiosis (Rocky Mountain Spotted Fever), Rubella (German Measles), Salmonellosis gastroenteritis (Salmonella), Scabies, Scombroid, Sepsis, Severe Acute Respiratory Syndrome (SARS), Shigellosis gastroenteritis (Shigella), Smallpox, Staphyloccal Infection Methicillin-resistant (MRSA), Staphylococcal Food Poisoning Enterotoxin B Poisoning (Staph Food Poisoning), Saphylococcal Infection Vancomycin Intermediate (VISA), Staphylococcal Infection Vancomycin Resistant (VRSA), Streptococcal Disease Group A (invasive) (Strep A (invasive), Streptococcal Disease, Group B (Strep-B), Streptococcal Toxic-Shock Syndrome STSS Toxic Shock, Syphilis (primary, secondary, early latent, late latent, congenital), Tetanus Infection, Trichomoniasis, Trichonosis Infection, Tuberculosis (TB), Tuberculosis Latent (LTBI), Tularemia, Typhoid Fever Group D, Vaginosis, Varicella (Chickenpox), Vibrio cholerae (Cholera), Vibriosis (Vibrio), Ebola Virus Hemorrhagic Fever, Lasa Virus Hemorrhagic Fever, Marburg Virus Hemorrhagic Fever, West Nile Virus, Yellow Fever, Yersenia, and Zika Virus Infection; the disease is associated with expression of a tumor-associated antigen, optionallythe disease associated with expression of a tumor antigen-associated is selected from the group consisting of a proliferative disease, a precancerous condition, a cancer, and a noncancer related indication associated with expression of the tumor antigen; the cancer is selected from the group consisting of colon cancer, rectal cancer, renal-cell carcinoma, liver cancer, non-small cell carcinoma of the lung, cancer of the small intestine, cancer of the esophagus, melanoma, bone cancer, pancreatic cancer, skin cancer, cancer of the head or neck, cutaneous or intraocular malignant melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, stomach cancer, testicular cancer, uterine cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, Hodgkin's Disease, non-Hodgkin lymphoma, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue, cancer of the urethra, cancer of the penis, solid tumors of childhood, cancer of the bladder, cancer of the kidney or ureter, carcinoma of the renal pelvis, neoplasm of the central nervous system (CNS), primary CNS lymphoma, tumor angiogenesis, spinal axis tumor, brain stem glioma, pituitary adenoma, Kaposi's sarcoma, epidermoid cancer, squamous cell cancer, T-cell lymphoma, environmentally induced cancers, combinations of said cancers, and metastatic lesions of said cancers; and / or the cancer is a hematologic cancer chosen from one or more of chronic lymphocytic leukemia (CLL), acute leukemias, acute lymphoid leukemia (ALL), B-cell acute lymphoid leukemia (B-ALL), T-cell acute lymphoid leukemia (T-ALL), chronic myelogenous leukemia (CML), B cell prolymphocytic leukemia, blastic plasmacytoid dendritic cell neoplasm, Burkitt's lymphoma, diffuse large B cell lymphoma, follicular lymphoma, hairy cell leukemia, small cell- or a large cell-follicular lymphoma, malignant lymphoproliferative conditions, MALT lymphoma, mantle cell lymphoma, marginal zone lymphoma, multiple myeloma, myelodysplasia and myelodysplastic syndrome, nonHodgkin's lymphoma, Hodgkin's lymphoma, plasmablastic lymphoma, plasmacytoid dendritic cell neoplasm, Waldenstrom macroglobulinemia, or pre-leukemia.

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