Pharmaceutical compositions and related methods for delivery of herpes simplex virus antigens - Patents.com

Abstract

The present disclosure provides pharmaceutical compositions and related technology (e.g., components thereof and / or methods related thereto) for delivery of HSV antigens (e.g., HSV vaccines). The present disclosure provides pharmaceutical compositions (e.g., immunogenic compositions, e.g., vaccines) and related technology (e.g., methods) for delivery of specific herpes simplex virus (HSV) antigen constructs (e.g., HSV-1 antigen constructs, HSV-2 antigen constructs, or combinations thereof) to a subject (e.g., a patient).

Description

[Background technology]

[0001] Herpes simplex virus (HSV), commonly referred to simply as herpes, is classified into two types: herpes simplex virus type 1 (HSV-1, or oral herpes) and herpes simplex virus type 2 (HSV-2, or genital herpes). According to the World Health Organization, an estimated 3.7 billion people under the age of 50 worldwide (67% of the world's population) are infected with HSV-1. The prevalence of HSV-1 is understood to be highest in Africa and lowest in the Americas. An estimated 491 million people aged 15–49 worldwide (13% of the world's population) are infected with HSV-2. More women than men are infected with HSV-2 because sexual transmission of HSV is more efficient from male to female than from female to male. The prevalence of HSV-2 infection is estimated to be highest in Africa, followed by the Americas. While HSV-2 prevalence has also been shown to increase with age, the number of newly infected people has historically been highest among adolescents. Both HSV-1 and HSV-2 infections are lifelong. Summary of the Invention [Means for solving the problem]

[0002] The present disclosure provides pharmaceutical compositions (e.g., immunogenic compositions, e.g., vaccines) and related technologies (e.g., methods) for delivering certain herpes simplex virus (HSV) antigen constructs (e.g., HSV-1 antigen constructs, HSV-2 antigen constructs, or combinations thereof) to a subject (e.g., a patient). In particular, the present disclosure provides HSV (e.g., HSV-1, HSV-2, or both) vaccine compositions and related technologies (e.g., methods). The present disclosure includes the unexpected discovery that the HSV antigens and antigenic fragments thereof provided in Tables 3-5 below are particularly advantageous for use in the prevention or treatment of HSV, for example, in the HSV antigen constructs and / or HSV vaccines further disclosed herein.

[0003] The present disclosure provides, for example, polyribonucleotides encoding one or more HSV antigens (e.g., HSV-1 antigens, HSV-2 antigens, or combinations thereof) or antigenic fragments thereof. In some embodiments, such polyribonucleotides can be part of an RNA construct. In some embodiments, the polyribonucleotides or RNA constructs described herein can be part of a composition (e.g., a pharmaceutical composition, e.g., an immunogenic composition, e.g., a vaccine).

[0004] The present disclosure provides a polyribonucleotide encoding a polypeptide. In some embodiments, the polypeptide comprises one or more herpes simplex virus (HSV) antigens or antigenic fragments thereof.

[0005] In some embodiments, the one or more HSV antigens or antigenic fragments thereof comprise (i) an HSV-1 antigen or antigenic fragment thereof, (ii) an HSV-2 antigen or antigenic fragment thereof, or (iii) a combination thereof.

[0006] In some embodiments, the polypeptide comprises a single HSV antigen or antigenic fragment thereof. In some embodiments, the polypeptide comprises a single HSV antigen. In some embodiments, the polypeptide comprises a single HSV antigenic fragment.

[0007] In some embodiments, the polypeptide comprises two or more HSV antigens or antigenic fragments thereof. In some embodiments, the polypeptide comprises two or more HSV antigens. In some embodiments, the polypeptide comprises two or more HSV antigenic fragments, wherein the two or more HSV antigenic fragments are each fragments of different HSV antigens. In some embodiments, the polypeptide comprises two or more HSV antigenic fragments, wherein at least two of the HSV antigenic fragments are fragments derived from the same HSV antigen. In some embodiments, the polypeptide comprises three or more HSV antigens or antigenic fragments thereof. In some embodiments, the polypeptide comprises four or more HSV antigens or antigenic fragments thereof.

[0008] In some embodiments, the polypeptide does not include a full-length HSV antigen.

[0009] In some embodiments, the one or more HSV antigens or antigenic fragments thereof comprise one or more T cell antigens or antigenic fragments thereof. In some embodiments, the one or more HSV antigens or antigenic fragments thereof comprise one or more B cell antigens or antigenic fragments thereof.

[0010] In some embodiments, the one or more HSV antigens or antigenic fragments thereof have at least 80% sequence identity, e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity, to one or more sequences or antigenic fragments thereof selected from SEQ ID NOs: 1-74. In some embodiments, the polypeptide comprises one or more HSV-2 antigens or antigenic fragments thereof comprising or consisting of an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to an amino acid sequence selected from SEQ ID NOs: 174-196.

[0011] In some embodiments, the one or more HSV antigens or antigenic fragments thereof are selected from the group consisting of: (i) one or more HSV RS1 polypeptides or antigenic fragments thereof, (ii) one or more HSV RL2 polypeptides or antigenic fragments thereof, (iii) one or more HSV UL1 polypeptides or antigenic fragments thereof, (iv) one or more HSV UL5 polypeptides or antigenic fragments thereof, (v) one or more HSV UL9 polypeptides or antigenic fragments thereof, (vi) one or more HSV UL19 polypeptides or antigenic fragments thereof, (vii) one or more HSV UL21 polypeptides or antigenic fragments thereof, (viii) one or more HSV UL25 polypeptides or antigenic fragments thereof, (ix) one or more HSV UL27 polypeptides or antigenic fragments thereof, (x) one or more HSV UL29 polypeptides or antigenic fragments thereof, (xi) one or more HSV UL30 polypeptides or antigenic fragments thereof, (xii) one or more HSV UL39 polypeptides or antigenic fragments thereof, (xiii) one or more HSV UL40 polypeptides or antigenic fragments thereof, (xiv) one or more HSV (xv) one or more HSV UL46 polypeptides or antigenic fragments thereof, (xv) one or more HSV UL47 polypeptides or antigenic fragments thereof, (xvi) one or more HSV UL48 polypeptides or antigenic fragments thereof, (xvii) one or more HSV UL49 polypeptides or antigenic fragments thereof, (xviii) one or more HSV UL52 polypeptides or antigenic fragments thereof, (xix) one or more HSV UL54 polypeptides or antigenic fragments thereof, or (xx) a combination thereof.In some embodiments, the polypeptide comprises one or more HSV antigenic fragments, the one or more HSV antigenic fragments being (i) one or more HSV RS1 polypeptide antigenic fragments, (ii) one or more HSV RL2 polypeptide antigenic fragments, (iii) one or more HSV UL1 polypeptide antigenic fragments, (iv) one or more HSV UL5 polypeptide antigenic fragments, (v) one or more HSV UL9 polypeptide antigenic fragments, (vi) one or more HSV UL19 polypeptide antigenic fragments, (vii) one or more HSV UL21 polypeptide antigenic fragments, (viii) one or more HSV UL25 polypeptide antigenic fragments, (ix) one or more HSV UL27 polypeptide antigenic fragments, (x) one or more HSV UL29 polypeptide antigenic fragments, (xi) one or more HSV UL30 polypeptide antigenic fragments, (xii) one or more HSV UL39 polypeptide antigenic fragments, (xiii) one or more HSV UL40 polypeptide antigenic fragments, (xiv) one or more HSV UL46 polypeptide antigenic fragments, (xv) one or more HSV (xvi) one or more HSV UL47 polypeptide antigenic fragments, (xvii) one or more HSV UL49 polypeptide antigenic fragments, (xviii) one or more HSV UL52 polypeptide antigenic fragments, (xix) one or more HSV UL54 polypeptide antigenic fragments, or (xx) a combination thereof.

[0012] In some embodiments, the polypeptide comprises one or more HSV RL2 polypeptides or antigenic fragments thereof, one or more HSV RS1 polypeptides or antigenic fragments thereof, and one or more HSV UL54 polypeptides or antigenic fragments thereof. In some embodiments, the polypeptide comprises an HSV-1 gD secretory signal, one or more RL2 polypeptides or antigenic fragments thereof, one or more RS1 polypeptides or antigenic fragments thereof, one or more UL54 polypeptides or antigenic fragments thereof, and an MITD.

[0013] In some embodiments, the polypeptide comprises, in order from N-terminus to C-terminus, an HSV-1 gD secretory signal, an RL2 polypeptide or an antigenic fragment thereof, a linker, an RL2 polypeptide or an antigenic fragment thereof, a linker, an RS1 polypeptide or an antigenic fragment thereof, a linker, a UL54 polypeptide or an antigenic fragment thereof, a linker, and a nucleotide sequence encoding MITD. In some embodiments, the polypeptide comprises or consists of an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence of SEQ ID NO:197.

[0014] In some embodiments, the polypeptide comprises a nucleotide sequence encoding, in N-terminal to C-terminal order, an HSV-1 gD secretory signal, a UL54 polypeptide or antigenic fragment thereof, a linker, an RS1 polypeptide or antigenic fragment thereof, a linker, an RL2 polypeptide or antigenic fragment thereof, a linker, an RL2 polypeptide or antigenic fragment thereof, a linker, and an MITD. In some embodiments, the polypeptide comprises or consists of an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence according to SEQ ID NO:201.

[0015] In some embodiments, the polypeptide comprises, in order from N-terminus to C-terminus, an HSV-2 gD secretory signal, an RL2 polypeptide or antigenic fragment thereof, a linker, an RL2 polypeptide or antigenic fragment thereof, a linker, an RS1 polypeptide or antigenic fragment thereof, a linker, a UL54 polypeptide or antigenic fragment thereof, a linker, and a nucleotide sequence encoding MITD. In some embodiments, the polypeptide comprises or consists of an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to an amino acid sequence according to SEQ ID NO:205.

[0016] In some embodiments, the polypeptide comprises one or more HSV UL29 polypeptides or antigenic fragments thereof, one or more HSV UL39 polypeptides or antigenic fragments thereof, one or more HSV UL49 polypeptides or antigenic fragments thereof, and one or more HSV UL9 polypeptides or antigenic fragments thereof. In some embodiments, the polypeptide comprises an HSV-1 gD secretory signal, one or more HSV UL29 polypeptides or antigenic fragments thereof, one or more HSV UL39 polypeptides or antigenic fragments thereof, one or more HSV UL49 polypeptides or antigenic fragments thereof, one or more HSV UL9 polypeptides or antigenic fragments thereof, and an MITD.

[0017] In some embodiments, the polypeptide comprises, in order from N-terminus to C-terminus, a nucleotide sequence encoding an HSV-1 gD secretory signal, a UL29 polypeptide or antigenic fragment thereof, a linker, a UL39 polypeptide or antigenic fragment thereof, a linker, a UL49 polypeptide or antigenic fragment thereof, a linker, a UL9 polypeptide or antigenic fragment thereof, a linker, and a MITD. In some embodiments, the polypeptide comprises or consists of an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence according to SEQ ID NO:198.

[0018] In some embodiments, the polypeptide comprises, in order from N-terminus to C-terminus, a nucleotide sequence encoding an HSV-1 gD secretory signal, a UL9 polypeptide or antigenic fragment thereof, a linker, a UL49 polypeptide or antigenic fragment thereof, a linker, a UL39 polypeptide or antigenic fragment thereof, a linker, a UL29 polypeptide or antigenic fragment thereof, a linker, and a MITD. In some embodiments, the polypeptide comprises or consists of an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to an amino acid sequence according to SEQ ID NO:202.

[0019] In some embodiments, the polypeptide comprises one or more HSV UL30 polypeptides or antigenic fragments thereof, one or more HSV UL40 polypeptides or antigenic fragments thereof, one or more HSV UL5 polypeptides or antigenic fragments thereof, and one or more HSV UL52 polypeptides or antigenic fragments thereof. In some embodiments, the polypeptide comprises an HSV-1 gD secretory signal, one or more HSV UL30 polypeptides or antigenic fragments thereof, one or more HSV UL40 polypeptides or antigenic fragments thereof, one or more HSV UL5 polypeptides or antigenic fragments thereof, one or more HSV UL52 polypeptides or antigenic fragments thereof, and an MITD.

[0020] In some embodiments, the polypeptide comprises, in order from N-terminus to C-terminus, a nucleotide sequence encoding an HSV-1 gD secretory signal, a UL30 polypeptide or antigenic fragment thereof, a linker, a UL30 polypeptide or antigenic fragment thereof, a linker, a UL40 polypeptide or antigenic fragment thereof, a linker, a UL5 polypeptide or antigenic fragment thereof, a linker, a UL5 polypeptide or antigenic fragment thereof, a linker, a UL52 polypeptide or antigenic fragment thereof, a linker, and a MITD. In some embodiments, the polypeptide comprises or consists of an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence according to SEQ ID NO:199.

[0021] In some embodiments, the polypeptide comprises, in order from N-terminus to C-terminus, a nucleotide sequence encoding an HSV-1 gD secretory signal, a UL52 polypeptide or antigenic fragment thereof, a linker, a UL5 polypeptide or antigenic fragment thereof, a linker, a UL5 polypeptide or antigenic fragment thereof, a linker, a UL40 polypeptide or antigenic fragment thereof, a linker, a UL30 polypeptide or antigenic fragment thereof, a linker, a UL30 polypeptide or antigenic fragment thereof, a linker, and a MITD. In some embodiments, the polypeptide comprises or consists of an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence according to SEQ ID NO:203.

[0022] In some embodiments, the polypeptide comprises one or more HSV UL1 polypeptides or antigenic fragments thereof, one or more HSV UL19 polypeptides or antigenic fragments thereof, one or more HSV UL21 polypeptides or antigenic fragments thereof, one or more HSV UL27 polypeptides or antigenic fragments thereof, one or more HSV UL46 polypeptides or antigenic fragments thereof, one or more HSV UL47 polypeptides or antigenic fragments thereof, one or more UL48 polypeptides or antigenic fragments thereof, and one or more HSV UL25 polypeptides or antigenic fragments thereof. In some embodiments, the polypeptide comprises an HSV-1 gD secretory signal, one or more HSV UL1 polypeptides or antigenic fragments thereof, one or more HSV UL19 polypeptides or antigenic fragments thereof, one or more HSV UL21 polypeptides or antigenic fragments thereof, one or more HSV UL27 polypeptides or antigenic fragments thereof, one or more HSV UL46 polypeptides or antigenic fragments thereof, one or more HSV UL47 polypeptides or antigenic fragments thereof, one or more UL48 polypeptides or antigenic fragments thereof, one or more HSV UL25 polypeptides or antigenic fragments thereof, and an MITD.

[0023] In some embodiments, the polypeptide comprises, in N-terminal to C-terminal order, an HSV-1 gD secretory signal, an HSV UL1 polypeptide or antigenic fragment thereof, a linker, an HSV UL19 polypeptide or antigenic fragment thereof, a linker, an HSV UL21 polypeptide or antigenic fragment thereof, a linker, an HSV UL27 polypeptide or antigenic fragment thereof, a linker, an HSV UL27 polypeptide or antigenic fragment thereof, a linker, an HSV UL46 polypeptide or antigenic fragment thereof, a linker, an HSV UL47 polypeptide or antigenic fragment thereof, a linker, an HSV UL25 polypeptide or antigenic fragment thereof, a linker, an HSV UL48 polypeptide or antigenic fragment thereof, a linker, and a nucleotide sequence encoding MITD. In some embodiments, the polypeptide comprises or consists of an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence according to SEQ ID NO:200.

[0024] In some embodiments, the polypeptide comprises, from N-terminus to C-terminus, an HSV-1 gD secretory signal, an HSV UL48 polypeptide or antigenic fragment thereof, a linker, an HSV UL25 polypeptide or antigenic fragment thereof, a linker, an HSV UL47 polypeptide or antigenic fragment thereof, a linker, an HSV UL46 polypeptide or antigenic fragment thereof, a linker, an HSV UL27 polypeptide or antigenic fragment thereof, a linker, an HSV UL27 polypeptide or antigenic fragment thereof, a linker, an HSV UL21 polypeptide or antigenic fragment thereof, a linker, an HSV UL19 polypeptide or antigenic fragment thereof, a linker, an HSV UL1 polypeptide or antigenic fragment thereof, a linker, and a nucleotide sequence encoding MITD. In some embodiments, the polypeptide comprises or consists of an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence according to SEQ ID NO:204.

[0025] In some embodiments, the one or more HSV antigens or antigenic fragments thereof comprise one or more HSV glycoproteins, hi some embodiments, the one or more HSV glycoproteins comprise HSV glycoprotein B (gB), HSV glycoprotein E (gE), HSV glycoprotein G (gG), HSV glycoprotein H (gH), HSV glycoprotein I (gI), HSV glycoprotein L (gL), or a combination thereof.

[0026] In some embodiments, the polypeptide comprises a single HSV antigen. In some embodiments, the single HSV antigen is an HSV glycoprotein. In some embodiments, the HSV glycoprotein is a full-length HSV glycoprotein. In some embodiments, the HSV glycoprotein is HSV gB, HSV gE, HSV gG, HSV gH, HSV gI, and HSV gL.

[0027] In some embodiments, the HSV glycoprotein is HSV-2 gB. In some embodiments, HSV-2 gB is or comprises an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 7, 8, 9, or 74. In some embodiments, HSV-2 gB comprises or consists of an amino acid sequence according to SEQ ID NO: 7, 8, 9, or 74.

[0028] In some embodiments, the HSV glycoprotein is HSV-2 gE. In some embodiments, the HSV-2 gE is or comprises an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 66, 67, 68, or 69. In some embodiments, the HSV-2 gE comprises or consists of an amino acid sequence according to SEQ ID NO: 66, 67, 68, or 69. In some embodiments, a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 80, 81, 82, 83, or 84.

[0029] In some embodiments, the HSV glycoprotein is HSV-2 gH. In some embodiments, the HSV-2 gH is or comprises an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 70, 71, 72, or 74. In some embodiments, the HSV-2 gH consists of or comprises an amino acid sequence according to SEQ ID NO: 70, 71, 72, or 74.

[0030] In some embodiments, the HSV glycoprotein is HSV-2 gI. In some embodiments, the HSV-2 gI is or comprises an amino acid sequence that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 62, 63, 64, or 65. In some embodiments, the HSV-2 gI comprises or consists of an amino acid sequence according to SEQ ID NO: 62, 63, 64, or 65. In some embodiments, the sequence is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 75, 76, 77, 78, or 79.

[0031] In some embodiments, the HSV glycoprotein is HSV-2 gL. In some embodiments, the HSV-2 gL is or comprises an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 58, 59, 60, or 61. In some embodiments, the HSV-2 gL comprises or consists of an amino acid sequence according to SEQ ID NO: 58, 59, 60, or 61.

[0032] In some embodiments, the polypeptide comprises a secretory signal. In some embodiments, the secretory signal comprises or consists of a viral secretory signal. In some embodiments, the viral secretory signal comprises or consists of an HSV secretory signal. In some embodiments, the secretory signal is a heterologous secretory signal. In some embodiments, the HSV secretory signal comprises or consists of an HSV-1 or HSV-2 secretory signal.

[0033] In some embodiments, the HSV secretory signal is selected from a) a gD2 secretory signal, b) a gD1 secretory signal, c) a gB1 secretory signal, d) a gI2 secretory signal, e) a gE2 secretory signal, f) a gC2 secretory signal, g) an Eboz secretory signal, h) an IL2 secretory signal, and i) an HLA-DR secretory signal.

[0034] In some embodiments, the HSV secretion signal comprises or consists of an HSV gD secretion signal. In some embodiments, the HSV gD secretion signal comprises or consists of an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 87. In some embodiments, the HSV gD secretion signal comprises or consists of an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to SEQ ID NO: 88. In some embodiments, the HSV gD secretion signal consists of an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to SEQ ID NO: 110. In some embodiments, the HSV gD secretory signal consists of an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to SEQ ID NO:111.

[0035] In some embodiments, the secretory signal is located at the N-terminus of the polypeptide.

[0036] In some embodiments, the HSV secretory signal comprises or consists of the HSV-2 glycoprotein I (gI) secretory signal.

[0037] In some embodiments, the HSV-2 gI secretion signal comprises an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to SEQ ID NO:107.

[0038] In some embodiments, the HSV-2 gI secretion signal comprises an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to SEQ ID NO:108.

[0039] In some embodiments, the polypeptide comprises a transmembrane region. In some embodiments, the transmembrane region comprises or consists of a viral transmembrane region. In some embodiments, the transmembrane region comprises or consists of an HSV transmembrane region. In some embodiments, the HSV transmembrane region comprises or consists of an HSV-1 or HSV-2 transmembrane region. In some embodiments, the HSV transmembrane region comprises or consists of an HSV gD transmembrane region. In some embodiments, the HSV gD transmembrane region consists of an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 160.

[0040] In some embodiments, the polypeptide does not include a transmembrane region.

[0041] In some embodiments, the polypeptide comprises a multimerization domain. In some embodiments, the polypeptide comprises one or more linkers. The polyribonucleotide of item 215, wherein the one or more linkers comprise one or more glycine (G) residues and / or one or more serine (S) residues. In some embodiments, the one or more linkers comprise or consist of an amino acid sequence according to SEQ ID NO: 163. In some embodiments, the one or more linkers comprise or consist of an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 165. In some embodiments, the one or more linkers comprise or consist of an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 168. In some embodiments, one or more linkers comprise or consist of an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to SEQ ID NO:217.

[0042] In some embodiments, the polyribonucleotide is an isolated polyribonucleotide.

[0043] In some embodiments, the polyribonucleotide is an engineered polyribonucleotide.

[0044] In some embodiments, the polyribonucleotide is a codon-optimized polyribonucleotide.

[0045] The present disclosure also provides an RNA construct.

[0046] In some embodiments, an RNA construct comprises, in 5' to 3' order, (i) a 5' UTR, (ii) any polyribonucleotide according to the present disclosure, (iv) a 3' UTR, and (v) a poly-A tail sequence. In some embodiments, the RNA construct comprises (i) a 5' UTR that comprises or consists of a modified human alpha globin 5'-UTR, (ii) a 3' UTR that comprises or consists of a first sequence from the amino-terminal enhancer of a split (AES) messenger RNA and a second sequence from a mitochondrially encoded 12S ribosomal RNA, or (iii) both.

[0047] In some embodiments, the 5' UTR comprises or consists of a ribonucleic acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 208. In some embodiments, the 5' UTR comprises or consists of a ribonucleic acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 209. In some embodiments, the 3' UTR comprises or consists of a ribonucleic acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 215. In some embodiments, the 3'UTR comprises or consists of a ribonucleic acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 216. In some embodiments, the polyA tail sequence is a split polyA tail sequence. In some embodiments, the split polyA tail sequence consists of a ribonucleic acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to a ribonucleic acid sequence selected from SEQ ID NOs: 210, 212, or 213. In some embodiments, the RNA construct further comprises a 5' cap. In some embodiments, the RNA construct comprises a cap-proximal sequence that includes the +1, +2, +3, +4, and +5 positions of the polyribonucleotide. In some embodiments, the 5' cap comprises or consists of m7(3'OMeG)(5')ppp(5')(2'OMeA1)pG2, where A1 is the +1 position of the polyribonucleotide and G2 is the +2 position of the polyribonucleotide.In some embodiments, the cap-proximal sequence comprises a sequence comprising A3A4U5 (SEQ ID NO: 207) at A1 and G2 of the Cap 1 structure and at positions +3, +4, and +5 of the polyribonucleotide, respectively.

[0048] In some embodiments, the polyribonucleotide comprises modified uridines in place of every uridine, and optionally, each modified uridine is N1-methyl-pseudouridine.

[0049] The present disclosure also provides compositions.

[0050] In some embodiments, the composition comprises one or more polyribonucleotides according to the present disclosure. In some embodiments, the composition comprises one or more RNA constructs described in any one of paragraphs 224 to 236. In some embodiments, the composition further comprises a lipid nanoparticle, a polyplex (PLX), a lipidated polyplex (LPLX), or a liposome, wherein the one or more polyribonucleotides are fully or partially encapsulated within the lipid nanoparticle, the polyplex (PLX), the lipidated polyplex (LPLX), or the liposome. In some embodiments, the composition further comprises a lipid nanoparticle, wherein the one or more polyribonucleotides are encapsulated within the lipid nanoparticle.

[0051] The present disclosure also provides pharmaceutical compositions.

[0052] In some embodiments, a pharmaceutical composition comprises a composition according to the present disclosure and at least one pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical comprises a cryoprotectant, and optionally, the cryoprotectant is sucrose. In some embodiments, the pharmaceutical comprises an aqueous buffer solution, and optionally, the aqueous buffer solution comprises one or more of Tris base, Tris-HCl, NaCl, KCl, NaHPO, and KHPO.

[0053] The present disclosure also provides combinations.

[0054] In some embodiments, the combination comprises a first polyribonucleotide according to the present disclosure and a second polyribonucleotide according to the present disclosure, wherein the first polyribonucleotide and the second polyribonucleotide are different.

[0055] In some embodiments, the combination comprises a first pharmaceutical composition comprising a first polyribonucleotide, wherein the first polyribonucleotide is a polyribonucleotide according to the present disclosure, and a second pharmaceutical composition comprising a second polyribonucleotide, wherein the second polyribonucleotide is a polyribonucleotide according to the present disclosure, wherein the first polyribonucleotide and the second polyribonucleotide are different.

[0056] In some embodiments, the combination comprises a first polyribonucleotide according to the present disclosure and a second polyribonucleotide encoding a second polypeptide, wherein the second polypeptide comprises one or more HSV RL2 polypeptides or antigenic fragments thereof, one or more HSV RS1 polypeptides or antigenic fragments thereof, and one or more HSV UL54 polypeptides or antigenic fragments thereof. In some embodiments, the combination comprises a first pharmaceutical composition comprising a first polyribonucleotide, wherein the first polyribonucleotide is a polyribonucleotide according to the present disclosure, and a second pharmaceutical composition comprising a second polyribonucleotide, wherein the second polyribonucleotide comprises one or more HSV RL2 polypeptides or antigenic fragments thereof, one or more HSV RS1 polypeptides or antigenic fragments thereof, and one or more HSV UL54 polypeptides or antigenic fragments thereof. In some embodiments, the second polypeptide comprises, in order from N-terminus to C-terminus, an HSV-1 gD secretion signal, an RL2 polypeptide or antigenic fragment thereof, a linker, an RL2 polypeptide or antigenic fragment thereof, a linker, an RS1 polypeptide or antigenic fragment thereof, a linker, a UL54 polypeptide or antigenic fragment thereof, a linker, and an MITD. In some embodiments, the second polypeptide comprises or consists of an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 197. In some embodiments, the second polypeptide comprises, in order from N-terminus to C-terminus, an HSV-1 gD secretion signal, a UL54 polypeptide or antigenic fragment thereof, a linker, an RS1 polypeptide or antigenic fragment thereof, a linker, an RL2 polypeptide or antigenic fragment thereof, a linker, an RL2 polypeptide or antigenic fragment thereof, a linker, and an MITD.In some embodiments, the second polypeptide comprises or consists of an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 201. In some embodiments, the second polypeptide comprises, from N-terminus to C-terminus, an HSV-2 gD secretion signal, an RL2 polypeptide or an antigenic fragment thereof, a linker, an RL2 polypeptide or an antigenic fragment thereof, a linker, an RS1 polypeptide or an antigenic fragment thereof, a linker, a UL54 polypeptide or an antigenic fragment thereof, a linker, and an MITD. In some embodiments, the second polypeptide comprises or consists of an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 205.

[0057] In some embodiments, the combination comprises a first polyribonucleotide according to the present disclosure and a second polyribonucleotide encoding a second polypeptide, wherein the second polypeptide comprises one or more HSV UL29 polypeptides or antigenic fragments thereof, one or more HSV UL39 polypeptides or antigenic fragments thereof, one or more HSV UL49 polypeptides or antigenic fragments thereof, and one or more HSV UL9 polypeptides or antigenic fragments thereof. In some embodiments, the combination comprises a first pharmaceutical composition comprising the first polyribonucleotide, wherein the first polyribonucleotide is a polyribonucleotide according to the present disclosure, and a second pharmaceutical composition comprising the second polyribonucleotide, wherein the second polypeptide comprises one or more HSV UL29 polypeptides or antigenic fragments thereof, one or more HSV UL39 polypeptides or antigenic fragments thereof, one or more HSV UL49 polypeptides or antigenic fragments thereof, and one or more HSV UL9 polypeptides or antigenic fragments thereof. In some embodiments, the second polypeptide comprises, from N-terminus to C-terminus, an HSV-1 gD secretion signal, a UL29 polypeptide or antigenic fragment thereof, a linker, a UL39 polypeptide or antigenic fragment thereof, a linker, a UL49 polypeptide or antigenic fragment thereof, a linker, a UL9 polypeptide or antigenic fragment thereof, a linker, and an MITD. In some embodiments, the second polypeptide comprises or consists of an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 198. In some embodiments, the second polypeptide comprises, from N-terminus to C-terminus, an HSV-1 gD secretion signal, a UL9 polypeptide or antigenic fragment thereof, a linker, a UL49 polypeptide or antigenic fragment thereof, a linker, a UL39 polypeptide or antigenic fragment thereof, a linker, a UL29 polypeptide or antigenic fragment thereof, a linker, and an MITD.In some embodiments, the second polypeptide comprises or consists of an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO:202.

[0058] In some embodiments, the combination comprises a first polyribonucleotide according to the present disclosure and a second polyribonucleotide encoding a second polypeptide, wherein the second polypeptide comprises one or more HSV UL30 polypeptides or antigenic fragments thereof, one or more HSV UL40 polypeptides or antigenic fragments thereof, one or more HSV UL5 polypeptides or antigenic fragments thereof, and one or more HSV UL52 polypeptides or antigenic fragments thereof. In some embodiments, the combination comprises a first pharmaceutical composition comprising a first polyribonucleotide, wherein the first polyribonucleotide is a polyribonucleotide according to the present disclosure, and a second pharmaceutical composition comprising a second polyribonucleotide, wherein the second polypeptide comprises one or more HSV UL30 polypeptides or antigenic fragments thereof, one or more HSV UL40 polypeptides or antigenic fragments thereof, one or more HSV UL5 polypeptides or antigenic fragments thereof, and one or more HSV UL52 polypeptides or antigenic fragments thereof. In some embodiments, the second polypeptide comprises, in order from N-terminus to C-terminus, an HSV-1 gD secretory signal, a UL30 polypeptide or antigenic fragment thereof, a linker, a UL30 polypeptide or antigenic fragment thereof, a linker, a UL40 polypeptide or antigenic fragment thereof, a linker, a UL5 polypeptide or antigenic fragment thereof, a linker, a UL5 polypeptide or antigenic fragment thereof, a linker, a UL52 polypeptide or antigenic fragment thereof, a linker, and an MITD. In some embodiments, the second polypeptide comprises or consists of an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO:199.In some embodiments, the second polypeptide comprises, in order from N-terminus to C-terminus, an HSV-1 gD secretory signal, a UL52 polypeptide or antigenic fragment thereof, a linker, a UL5 polypeptide or antigenic fragment thereof, a linker, a UL5 polypeptide or antigenic fragment thereof, a linker, a UL40 polypeptide or antigenic fragment thereof, a linker, a UL30 polypeptide or antigenic fragment thereof, a linker, a UL30 polypeptide or antigenic fragment thereof, a linker, and an MITD. In some embodiments, the second polypeptide comprises or consists of an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO:203.

[0059] In some embodiments, the combination comprises a first polyribonucleotide according to the present disclosure and a second polyribonucleotide encoding a second polypeptide, wherein the second polypeptide comprises one or more HSV UL1 polypeptides or antigenic fragments thereof, one or more HSV UL19 polypeptides or antigenic fragments thereof, one or more HSV UL21 polypeptides or antigenic fragments thereof, one or more HSV UL27 polypeptides or antigenic fragments thereof, one or more HSV UL46 polypeptides or antigenic fragments thereof, one or more HSV UL47 polypeptides or antigenic fragments thereof, one or more UL48 polypeptides or antigenic fragments thereof, and one or more HSV UL25 polypeptides or antigenic fragments thereof. In some embodiments, the combination comprises a first pharmaceutical composition comprising a first polyribonucleotide, wherein the first polyribonucleotide is a polyribonucleotide according to the present disclosure, and a second pharmaceutical composition comprising a second polyribonucleotide, wherein the second polypeptide comprises one or more HSV UL1 polypeptides or antigenic fragments thereof, one or more HSV UL19 polypeptides or antigenic fragments thereof, one or more HSV UL21 polypeptides or antigenic fragments thereof, one or more HSV UL27 polypeptides or antigenic fragments thereof, one or more HSV UL46 polypeptides or antigenic fragments thereof, one or more HSV UL47 polypeptides or antigenic fragments thereof, one or more UL48 polypeptides or antigenic fragments thereof, and one or more HSV UL25 polypeptides or antigenic fragments thereof.In some embodiments, the second polypeptide comprises, from N-terminus to C-terminus, an HSV-1 gD secretory signal, an HSV UL1 polypeptide or an antigenic fragment thereof, a linker, an HSV UL19 polypeptide or an antigenic fragment thereof, a linker, an HSV UL21 polypeptide or an antigenic fragment thereof, a linker, an HSV UL27 polypeptide or an antigenic fragment thereof, a linker, an HSV UL27 polypeptide or an antigenic fragment thereof, a linker, an HSV UL46 polypeptide or an antigenic fragment thereof, a linker, an HSV UL47 polypeptide or an antigenic fragment thereof, a linker, an HSV UL25 polypeptide or an antigenic fragment thereof, a linker, an HSV UL48 polypeptide or an antigenic fragment thereof, a linker, and an MITD. In some embodiments, the second polypeptide comprises or consists of an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO:200. In some embodiments, the second polypeptide comprises, from N-terminus to C-terminus, an HSV-1 gD secretory signal, an HSV UL48 polypeptide or an antigenic fragment thereof, a linker, an HSV UL25 polypeptide or an antigenic fragment thereof, a linker, an HSV UL47 polypeptide or an antigenic fragment thereof, a linker, an HSV UL46 polypeptide or an antigenic fragment thereof, a linker, an HSV UL27 polypeptide or an antigenic fragment thereof, a linker, an HSV UL27 polypeptide or an antigenic fragment thereof, a linker, an HSV UL21 polypeptide or an antigenic fragment thereof, a linker, an HSV UL19 polypeptide or an antigenic fragment thereof, a linker, an HSV UL1 polypeptide or an antigenic fragment thereof, a linker, and an MITD. In some embodiments, the second polypeptide comprises or consists of an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 204. In some embodiments, the second polypeptide is HSV gB.In some embodiments, the second polypeptide consists of or comprises an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 7, 8, 9, or 74.

[0060] The present disclosure also provides a method comprising administering to a subject a polyribonucleotide according to the present disclosure, or an RNA construct according to the present disclosure.

[0061] The present disclosure also provides methods comprising administering a composition according to the present disclosure to a subject.

[0062] The present disclosure also provides methods that include administering to a subject one or more doses of a composition according to the present disclosure or a pharmaceutical composition according to the present disclosure.

[0063] The present disclosure also provides methods comprising administering a combination according to the present disclosure to a subject.

[0064] The present disclosure also provides a pharmaceutical composition according to the present disclosure for use in treating an HSV infection, comprising administering one or more doses of the pharmaceutical composition to a subject.

[0065] The present disclosure also provides a pharmaceutical composition according to the present disclosure for use in preventing HSV infection, comprising administering one or more doses of the pharmaceutical composition to a subject.

[0066] In some embodiments, a pharmaceutical composition according to the present disclosure for a method or use according to the present disclosure comprises administering two or more doses of the pharmaceutical composition to a subject.

[0067] In some embodiments, the method according to the present disclosure or the pharmaceutical composition for use according to the present disclosure comprises administering three or more doses of the pharmaceutical composition to the subject.

[0068] The present disclosure also provides methods comprising administering a combination according to the present disclosure to a subject. In some embodiments, the first pharmaceutical composition and the second pharmaceutical composition are administered on the same day. In some embodiments, the first pharmaceutical composition and the second pharmaceutical composition are administered on different days. In some embodiments, the first pharmaceutical composition and the second pharmaceutical composition are administered to the subject at different locations on the subject's body. In some embodiments, the method is a method for treating an HSV infection. In some embodiments, the method is a method for preventing an HSV infection. In some embodiments, the subject has or is at risk of developing an HSV infection.

[0069] In some embodiments, the subject is a human.

[0070] In some embodiments, administration induces an anti-HSV immune response in the subject. In some embodiments, the anti-HSV immune response in the subject comprises an adaptive immune response. In some embodiments, the anti-HSV immune response in the subject comprises a T cell response. In some embodiments, the T cell response is or comprises a CD4+ T cell response. In some embodiments, the T cell response is or comprises a CD8+ T cell response. In some embodiments, the anti-HSV immune system response comprises a B cell response. In some embodiments, the anti-HSV immune system response comprises the production of antibodies directed against one or more HSV antigens, or antigenic fragments thereof, that are at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to one or more sequences selected from SEQ ID NOs: 1-74, or antigenic fragments thereof.

[0071] The present disclosure also provides the use of a pharmaceutical composition according to the present disclosure in the treatment of herpes simplex virus infection.

[0072] The present disclosure also provides the use of a pharmaceutical composition according to the present disclosure in preventing herpes simplex virus infection.

[0073] The present disclosure also provides for the use of a pharmaceutical composition according to the present invention to induce an anti-herpes simplex immune virus response in a subject.

[0074] The present disclosure also provides polypeptides encoded by polyribonucleotides according to the present disclosure.

[0075] The present disclosure also provides a polypeptide encoded by the RNA construct of any one of items 224 to 236.

[0076] The present disclosure also provides a host cell comprising a polyribonucleotide according to the present disclosure.

[0077] The present disclosure also provides a host cell comprising an RNA construct according to the present disclosure.

[0078] The present disclosure also provides a host cell comprising a polypeptide according to the present disclosure. [Brief explanation of the drawings]

[0079] [Figure 1] Schematic diagram of an HSV particle. [Figure 2] Schematic diagram of the HSV life cycle. Modified from Ibanez, FJ, et al., "Experimental Dissection of the Lytic Replication Cycles of Herpes Simplex Virus in vitro," Front Microbiol. 2018;9:2406, which is incorporated herein by reference in its entirety. [Figure 3] Schematic diagram of a model of HSV latent infection. Modified from Knipe, DM, et al., "Clues to mechanisms of herpesviral latent infection and potential cures," PNAS September 29, 2015 112(39)11993-11994, which is incorporated herein by reference in its entirety. [Figure 4] 1 is a summary table of clinical trial results using HSV vaccine candidates. The table is modified from Aschner, CB, & Herold, BC (2021), Alphaherpesvirus vaccines. Current Issues in Molecular Biology, 41, 469-508, which is incorporated herein by reference in its entirety. [Figure 5] 1 is a summary table of HSV-2 vaccine candidates in preclinical development. The table is modified from Aschner, CB, & Herold, BC (2021), Alphaherpesvirus vaccines. Current Issues in Molecular Biology, 41, 469-508, which is incorporated herein by reference in its entirety. [Figure 6] 1 is a heat map assessing the phylogeny and homology of HSV-1 and HSV-2 genes. As shown, HSV-1 and HSV-2 genes are homologous with approximately 75% sequence identity. HSV-2 demonstrates minimal cross-strain variation. [Figure 7] Included are line graphs showing the time after HSV infection when immediate early, early, and late genes are expressed. [Figure 8] 1 is a table showing certain characteristics of the data analyzed from Hosken 2006, Jing 2012, and Long 2014, including HSV species, number of subjects, number of genes assayed, experimental method, and symptom status of subjects. [Figure 9-1] 1 is a graph showing the percentage of subjects in the data analyzed from Hosken 2006 who were determined to have T cells targeting the product of each of the 48 analyzed HSV genes at levels above the indicated threshold (greater than 20 SFU / 10). Data was extracted from the Hosken 2006 figure. [Figure 9-2] Same as above. [Figure 10-1]1 is a graph showing the percent of subjects in the data analyzed from Long2014 who were determined to have T cells and / or CD4+ T cells, respectively, that target the product of each of the 75 analyzed HSV genes. Data was extracted from the Long2014 figure. [Figure 10-2] Same as above. [Figure 11] This set of three graphs shows the correlation of T cells detected targeting each of a range of individual HSV genes between pairs of datasets analyzed from the literature, specifically between Hosken 2006 and Jing 2012, Hosken 2006 and Long 2014, or Jing 2012 and Long 2014. R values ​​are shown for each graph. Hosken 2006 / Jing 2012 correlations were observed despite the different species (Jing 2012 HSV-1, Hosken 2006 HSV-2). No correlation was observed between data from Long 2014 and either Hosken 2006 or Jing 2012. [Figure 12-1] 1 is a chart showing the expression levels for each of a range of HSV genes as determined from analysis of multiple data sets from diverse sources, including human cells, debris, and DRGs from latently infected mice. The dashed horizontal line indicates the determined median expression. [Figure 12-2] Same as above. [Figure 13] 12 is a chart plotting data from Hosken 2006 regarding the percentage of T cells targeting HSV gene products and the median expression for each of the various HSV genes (see FIG. 12). A threshold indicates genes that are both immunogenic and sufficiently expressed (upper right quadrant based on the dashed line indicating the threshold). [Figure 14] 12 is a chart plotting data from Jing 2012 regarding the percentage of T cells targeting HSV gene products and the median expression for each of the various HSV genes (see FIG. 12). A threshold indicates genes that are both immunogenic and sufficiently expressed (upper right quadrant based on the dashed line indicating the threshold). [Figure 15]Conservation scores determined for amino acids located at positions along the RL2 consensus sequence are shown. For this analysis, the complete HSV-1 and HSV-2 genomes were downloaded from the VIPR database, and HSV-1 strain 17 and HSV-2 strain HG52 were used as reference strains for HSV-1 and HSV-2, respectively. [Figure 16] Conservation scores determined for amino acids located at positions along the RS1 consensus sequence are shown. For this analysis, the complete HSV-1 and HSV-2 genomes were downloaded from the VIPR database, and HSV-1 strain 17 and HSV-2 strain HG52 were used as reference strains for HSV-1 and HSV-2, respectively. [Figure 17] Conservation scores determined for amino acids located at positions along the UL19 consensus sequence are shown. For this analysis, the complete HSV-1 and HSV-2 genomes were downloaded from the VIPR database, and HSV-1 strain 17 and HSV-2 strain HG52 were used as reference strains for HSV-1 and HSV-2, respectively. [Figure 18] Conservation scores determined for amino acids located at positions along the UL1 consensus sequence are shown. For this analysis, the complete HSV-1 and HSV-2 genomes were downloaded from the VIPR database, and HSV-1 strain 17 and HSV-2 strain HG52 were used as reference strains for HSV-1 and HSV-2, respectively. [Figure 19] Conservation scores determined for amino acids located at positions along the UL21 consensus sequence are shown. For this analysis, the complete HSV-1 and HSV-2 genomes were downloaded from the VIPR database, and HSV-1 strain 17 and HSV-2 strain HG52 were used as reference strains for HSV-1 and HSV-2, respectively. [Figure 20] Conservation scores determined for amino acids located at positions along the UL25 consensus sequence are shown. For this analysis, the complete HSV-1 and HSV-2 genomes were downloaded from the VIPR database, and HSV-1 strain 17 and HSV-2 strain HG52 were used as reference strains for HSV-1 and HSV-2, respectively. [Figure 21] Conservation scores determined for amino acids located at positions along the UL27 consensus sequence are shown. UL27 encodes HSV gB. For this analysis, complete HSV-1 and HSV-2 genomes were downloaded from the VIPR database, and HSV-1 strain 17 and HSV-2 strain HG52 were used as reference strains for HSV-1 and HSV-2, respectively. [Figure 22] Conservation scores determined for amino acids located at positions along the UL29 consensus sequence are shown. For this analysis, the complete HSV-1 and HSV-2 genomes were downloaded from the VIPR database, and HSV-1 strain 17 and HSV-2 strain HG52 were used as reference strains for HSV-1 and HSV-2, respectively. [Figure 23] Conservation scores determined for amino acids located at positions along the UL30 consensus sequence are shown. For this analysis, the complete HSV-1 and HSV-2 genomes were downloaded from the VIPR database, and HSV-1 strain 17 and HSV-2 strain HG52 were used as reference strains for HSV-1 and HSV-2, respectively. [Figure 24] Conservation scores determined for amino acids located at positions along the UL39 consensus sequence are shown. For this analysis, the complete HSV-1 and HSV-2 genomes were downloaded from the VIPR database, and HSV-1 strain 17 and HSV-2 strain HG52 were used as reference strains for HSV-1 and HSV-2, respectively. [Figure 25] Conservation scores determined for amino acids located at positions along the UL40 consensus sequence are shown. For this analysis, the complete HSV-1 and HSV-2 genomes were downloaded from the VIPR database, and HSV-1 strain 17 and HSV-2 strain HG52 were used as reference strains for HSV-1 and HSV-2, respectively. [Figure 26]Conservation scores determined for amino acids located at positions along the UL46 consensus sequence are shown. For this analysis, the complete HSV-1 and HSV-2 genomes were downloaded from the VIPR database, and HSV-1 strain 17 and HSV-2 strain HG52 were used as reference strains for HSV-1 and HSV-2, respectively. [Figure 27] Conservation scores determined for amino acids located at positions along the UL47 consensus sequence are shown. For this analysis, the complete HSV-1 and HSV-2 genomes were downloaded from the VIPR database, and HSV-1 strain 17 and HSV-2 strain HG52 were used as reference strains for HSV-1 and HSV-2, respectively. [Figure 28] Conservation scores determined for amino acids located at positions along the UL48 consensus sequence are shown. For this analysis, the complete HSV-1 and HSV-2 genomes were downloaded from the VIPR database, and HSV-1 strain 17 and HSV-2 strain HG52 were used as reference strains for HSV-1 and HSV-2, respectively. [Figure 29] Conservation scores determined for amino acids located at positions along the UL49 consensus sequence are shown. For this analysis, the complete HSV-1 and HSV-2 genomes were downloaded from the VIPR database, and HSV-1 strain 17 and HSV-2 strain HG52 were used as reference strains for HSV-1 and HSV-2, respectively. [Figure 30] Conservation scores determined for amino acids located at positions along the UL52 consensus sequence are shown. For this analysis, the complete HSV-1 and HSV-2 genomes were downloaded from the VIPR database, and HSV-1 strain 17 and HSV-2 strain HG52 were used as reference strains for HSV-1 and HSV-2, respectively. [Figure 31] Conservation scores determined for amino acids located at positions along the UL54 consensus sequence are shown. For this analysis, the complete HSV-1 and HSV-2 genomes were downloaded from the VIPR database, and HSV-1 strain 17 and HSV-2 strain HG52 were used as reference strains for HSV-1 and HSV-2, respectively. [Figure 32] Conservation scores determined for amino acids located at positions along the UL5 consensus sequence are shown. For this analysis, the complete HSV-1 and HSV-2 genomes were downloaded from the VIPR database, and HSV-1 strain 17 and HSV-2 strain HG52 were used as reference strains for HSV-1 and HSV-2, respectively. [Figure 33] Conservation scores determined for amino acids located at positions along the UL9 consensus sequence are shown. For this analysis, the complete HSV-1 and HSV-2 genomes were downloaded from the VIPR database, and HSV-1 strain 17 and HSV-2 strain HG52 were used as reference strains for HSV-1 and HSV-2, respectively. [Figure 34] Shown are HSV strain conservation scores determined for amino acids located at positions along the RL2 consensus sequence. [Figure 35] The HSV strain conservation scores determined for amino acids located at positions along the RS1 consensus sequence are shown. [Figure 36] Shown are HSV strain conservation scores determined for amino acids located at positions along the UL19 consensus sequence. [Figure 37] Shown are HSV strain conservation scores determined for amino acids located at positions along the UL1 consensus sequence. [Figure 38] Shown are HSV strain conservation scores determined for amino acids located at positions along the UL21 consensus sequence. [Figure 39] 1 shows the HSV strain conservation scores determined for amino acids located at positions along the UL25 consensus sequence. [Figure 40] 1 shows the HSV strain conservation scores determined for amino acids located at positions along the UL27 consensus sequence. [Figure 41] 1 shows the HSV strain conservation scores determined for amino acids located at positions along the UL29 consensus sequence. [Figure 42]Shown are HSV strain conservation scores determined for amino acids located at positions along the UL30 consensus sequence. [Figure 43] 1 shows the HSV strain conservation scores determined for amino acids located at positions along the UL39 consensus sequence. [Figure 44] Shown are HSV strain conservation scores determined for amino acids located at positions along the UL40 consensus sequence. [Figure 45] Shown are HSV strain conservation scores determined for amino acids located at positions along the UL46 consensus sequence. [Figure 46] Shown are HSV strain conservation scores determined for amino acids located at positions along the UL47 consensus sequence. [Figure 47] Shown are HSV strain conservation scores determined for amino acids located at positions along the UL48 consensus sequence. [Figure 48] 1 shows the HSV strain conservation scores determined for amino acids located at positions along the UL49 consensus sequence. [Figure 49] Shown are HSV strain conservation scores determined for amino acids located at positions along the UL52 consensus sequence. [Figure 50] Shown are HSV strain conservation scores determined for amino acids located at positions along the UL54 consensus sequence. [Figure 51] Shown are HSV strain conservation scores determined for amino acids located at positions along the UL5 consensus sequence. [Figure 52] Shown are HSV strain conservation scores determined for amino acids located at positions along the UL9 consensus sequence. [Figure 53]Four HSV antigen constructs, A, B, C, and D, are shown. Construct A contains the RL2, RL2, RS1, and UL54 T cell antigens. Construct B contains the UL29, UL39, UL49, and UL9 T cell antigens. Construct C contains the UL30, UL40, UL5, and UL52 T cell antigens. Construct D contains the UL1, UL19, UL21, UL27, UL46, UL47, UL25, and UL48 T cell antigens. DETAILED DESCRIPTION OF THE INVENTION

[0080] Specific Definitions Generally, terms used herein follow their understood meaning in the art unless expressly indicated otherwise. Explicit definitions of certain terms are provided below, and the meaning of these and other terms in specific instances throughout the specification will be apparent to those of ordinary skill in the art from the context.

[0081] In order that the present invention may be more readily understood, certain terms are first defined below. Further definitions of these and other terms are set forth throughout the specification.

[0082] About: The term "about," when used herein in reference to a value, refers to a value that is similar in relation to the referenced value. Generally, a person of ordinary skill in the art familiar with the context will understand the reasonable degree of variation encompassed by "about" in that context. For example, in some embodiments, the term "about" can encompass a range of values ​​within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less of the referenced value.

[0083] Agent: As used herein, the term "agent" may refer to a physical entity or phenomenon. In some embodiments, an agent may be characterized by a particular configuration and / or effect. In some embodiments, an agent may be a compound, molecule, or entity of any chemical class, including, for example, a small molecule, a polypeptide, a nucleic acid, a monosaccharide, a lipid, a metal, or a combination or complex thereof. In some embodiments, the term "agent" may refer to a compound, molecule, or entity that comprises a polymer. In some embodiments, the term may refer to a compound or entity that comprises one or more polymer moieties. In some embodiments, the term "agent" may refer to a compound, molecule, or entity that is substantially free of a particular polymer or polymer moiety. In some embodiments, the term may refer to a compound, molecule, or entity that is devoid of or substantially free of any polymers or polymer moieties.

[0084] Amino acid: In its broadest sense, the term "amino acid" as used herein refers to a compound and / or substance that can be, is, or is incorporated into a polypeptide chain, for example, by the formation of one or more peptide bonds. In some embodiments, an amino acid has the general structure HN-C(H)(R)-COOH. In some embodiments, an amino acid is a naturally occurring amino acid. In some embodiments, an amino acid is a non-naturally occurring amino acid; in some embodiments, an amino acid is a D-amino acid; in some embodiments, an amino acid is an L-amino acid. A "standard amino acid" refers to any of the 20 standard L-amino acids commonly found in naturally occurring peptides. A "non-standard amino acid" refers to any amino acid other than the standard amino acids, whether synthetically prepared or obtained from a natural source. In some embodiments, amino acids, including the carboxy-terminal amino acid and / or the amino-terminal amino acid in a polypeptide, may contain structural modifications compared to the above general structure. For example, in some embodiments, an amino acid may be modified relative to the general structure by methylation, amidation, acetylation, pegylation, glycosylation, phosphorylation, and / or substitution (e.g., of an amino group, a carboxylic acid group, one or more protons, and / or a hydroxyl group). In some embodiments, such modifications may, for example, alter the circulating half-life of a polypeptide containing the modified amino acid compared to one containing the same amino acid except for the unmodified amino acid. In some embodiments, such modifications do not significantly alter the relevant activity of a polypeptide containing the modified amino acid compared to one containing the same amino acid except for the unmodified amino acid. As will be clear from the context, in some embodiments, the term "amino acid" may be used to refer to a free amino acid; in some embodiments, the term may be used to refer to an amino acid residue of a polypeptide.

[0085] Antibody agent: As used herein, the term "antibody agent" refers to an agent that specifically binds to a particular antigen. In some embodiments, the term encompasses a polypeptide or polypeptide complex that contains sufficient immunoglobulin structural elements to confer specific binding. For example, in some embodiments, an antibody agent is or includes a polypeptide whose amino acid sequence includes one or more structural elements recognized by those skilled in the art as a complementarity-determining region (CDR); in some embodiments, an antibody agent is or includes a polypeptide that includes at least one CDR (e.g., at least one heavy chain CDR and / or at least one light chain CDR) whose amino acid sequence is substantially identical to that found in a reference antibody. In some embodiments, the included CDRs are substantially identical to the reference CDRs in that they are identical in sequence or contain one to five amino acid substitutions compared to the reference CDR. In some embodiments, the included CDR is substantially identical to the reference CDR in that it exhibits at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with the reference CDR. In some embodiments, the included CDR is substantially identical to the reference CDR in that it exhibits at least 96%, 96%, 97%, 98%, 99%, or 100% sequence identity with the reference CDR. In some embodiments, the included CDR is substantially identical to the reference CDR in that at least one amino acid within the included CDR is deleted, added, or substituted compared to the reference CDR, but the included CDR has an amino acid sequence that is otherwise identical to that of the reference CDR. In some embodiments, the included CDRs are substantially identical to the reference CDRs in that one to five amino acids within the included CDRs have been deleted, added, or substituted relative to the reference CDR, but the included CDRs have an amino acid sequence that is otherwise identical to that of the reference CDR. In some embodiments, the included CDRs are substantially identical to the reference CDRs in that at least one amino acid within the included CDRs has been substituted relative to the reference CDR, but the included CDRs have an amino acid sequence that is otherwise identical to that of the reference CDR.In some embodiments, the included CDR is substantially identical to the reference CDR in that one to five amino acids within the included CDR are deleted, added, or substituted relative to the reference CDR, but the included CDR has an amino acid sequence that is otherwise identical to the reference CDR. In some embodiments, the antibody agent is or comprises a polypeptide whose amino acid sequence comprises structural elements recognized by those skilled in the art as an immunoglobulin variable domain. In some embodiments, the antibody agent is or comprises a polypeptide whose amino acid sequence comprises structural elements recognized by those skilled in the art as corresponding to CDR1, 2, and 3 of an antibody variable domain; in some such embodiments, the antibody agent is or comprises a polypeptide or set of polypeptides whose amino acid sequence(s) together comprise structural elements recognized by those skilled in the art as corresponding to both heavy chain variable region CDRs and light chain variable region CDRs, e.g., heavy chain CDR1, 2, and / or 3, and light chain CDR1, 2, and / or 3. In some embodiments, the antibody agent is a polypeptide protein having a binding domain that is homologous or largely homologous to an immunoglobulin binding domain. In some embodiments, the antibody agent may be or comprise a polyclonal antibody preparation. In some embodiments, the antibody agent may be or comprise a monoclonal antibody preparation. In some embodiments, the antibody agent may comprise one or more constant region sequences unique to a particular organism, e.g., camel, human, mouse, primate, rabbit, rat; in many embodiments, the antibody agent may comprise one or more constant region sequences unique to humans. In some embodiments, the antibody agent may comprise one or more sequence elements recognized by those skilled in the art as humanized sequences, primatized sequences, chimeric sequences, etc. In some embodiments, the antibody agent may be a standard antibody (e.g., may comprise two heavy chains and two light chains).In some embodiments, antibody agents include, but are not limited to, intact IgA, IgG, IgE, or IgM antibodies; bispecific or multispecific antibodies (e.g., Zybodies®, etc.); antibody fragments, such as Fab fragments, Fab' fragments, F(ab')2 fragments, Fd' fragments, Fd fragments, and isolated CDRs or sets thereof; single chain Fvs; polypeptide-Fc fusions; single domain antibodies (e.g., shark single domain antibodies, e.g., IgNAR, or fragments thereof); camelid antibodies; masked antibodies (e.g., Probodies®); Small Modular The antibody may be in a format selected from ImmunoPharmaceuticals [SMIPs™]; single chain or Tandem diabodies [TandAb®]; VHH; Anticalins®; Nanobodies® minibodies; BiTEs®; ankyrin repeat proteins or DARPINs®; Avimers®; DARTs; TCR-like antibodies; Adnectins®; Affilins®; Trans-bodies®; Affibodies®; TrimerX®; MicroProteins; Fynomers®, Centyrins®; and KALBITOR®. In some embodiments, the antibody may lack covalent modifications (e.g., glycan attachments) that it would have if produced in nature. In some embodiments, the antibody may contain covalent modifications (e.g., glycan attachments, payloads (e.g., detectable moieties, therapeutic moieties, catalytic moieties, etc.), or other pendant groups (e.g., polyethylene glycol, etc.).

[0086] Antigen: Upon reading this specification, one of skill in the art will understand that the term "antigen" refers to a molecule that is recognized by the immune system, e.g., in certain embodiments, the adaptive immune system, to elicit an antigen-specific immune response. In some embodiments, the antigen-specific immune response can be or include the generation of antibodies and / or antigen-specific T cells. In some embodiments, the antigen is a peptide or polypeptide that includes at least one epitope to which an immune response can be generated. In one embodiment, the antigen is presented by a cell of the immune system, such as an antigen-presenting cell, such as a dendritic cell or macrophage. In one embodiment, the antigen, such as a T cell antigen, or its processed product is bound by a T cell or B cell receptor, or by an immunoglobulin molecule, such as an antibody. Thus, the antigen or its processed product can specifically react with an antibody or T lymphocyte (T cell). In one embodiment, the antigen is a parasitic antigen. According to the present disclosure, in some embodiments, the antigen may be delivered by an RNA molecule, as described herein. In some embodiments, a peptide or polypeptide antigen can be between 2 and 100 amino acids, including, for example, 5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 amino acids in length. In some embodiments, a peptide or polypeptide antigen can be more than 50 amino acids. In some embodiments, a peptide or polypeptide antigen can be more than 100 amino acids. In some embodiments, an antigen is recognized by an immune effector cell. In some embodiments, when recognized by an immune effector cell, the antigen, in the presence of an appropriate costimulatory signal, can induce stimulation, priming, and / or expansion of immune effector cells bearing an antigen receptor that recognizes the antigen. In the context of embodiments of the present disclosure, in some embodiments, an antigen can be presented or present on the surface of a cell, e.g., an antigen-presenting cell. In one embodiment, the antigen is presented by a diseased cell, such as a virus-infected cell. In one embodiment, the antigen receptor is a TCR that binds to an epitope of an antigen presented in the context of an MHC.In one embodiment, binding of the TCR, when expressed by and / or present on a T cell, to an antigen presented by a cell, such as an antigen-presenting cell, results in stimulation, priming, and / or expansion of the T cell. In one embodiment, binding of the TCR, when expressed by and / or present on a T cell, to an antigen presented on a diseased cell results in cytolysis and / or apoptosis of the diseased cell, and the T cell preferably releases cytotoxic factors, such as perforin and granzymes.

[0087] Associated: Two events or entities are "associated" with one another, as this term is used herein, when the presence, level, degree, type, and / or form of one correlates with that of the other. For example, a particular entity (e.g., a polypeptide, genetic signature, metabolite, microorganism, etc.) is considered associated with a particular disease, disorder, or condition if its presence, level, and / or form correlates with the occurrence, susceptibility, severity, stage, etc. of the disease, disorder, or condition (e.g., across a relevant population). In some embodiments, two or more entities are physically "associated" with one another if they interact directly or indirectly to be in physical proximity and / or remain in close proximity to one another. In some embodiments, two or more entities that are physically associated with one another are covalently linked to one another. In some embodiments, two or more entities that are physically associated with one another are not covalently linked to one another, but are non-covalently associated, for example, by hydrogen bonding, van der Waals interactions, hydrophobic interactions, magnetism, and combinations thereof.

[0088] Binding: Upon reading this specification, one of skill in the art will understand that the term "binding" typically refers to a non-covalent association between entities or moieties. In some embodiments, binding data is expressed as "IC50." As understood in the art, IC50 is the concentration of an agent being evaluated in a binding assay at which 50% inhibition of binding of a reference agent known to bind to the relevant binding partner is observed. In some embodiments, the assay is performed under conditions (e.g., limiting binding target and reference concentrations) at which the assay is performed, and these values ​​are expressed as K D The binding assay is well known in the art and is described in detail in, for example, PCT Publications WO94 / 20127 and WO94 / 03205, as well as in other publications such as, for example, Sidney, et al., Current Protocols in Immunology, 18.3.1 (1998); Sidney, et al., J. Immunol. 154:247 (1995); and Sette, et al., Mol. Immunol. 31:813 (1994). Alternatively, binding can be expressed relative to binding by a reference standard peptide. For example, the IC of the reference standard peptide 50 for that IC 50Binding can be based on live cells (e.g., Ceppellini et al., Nature, 339:392 (1989); Christnick et al., Nature, 352:67 (1991); Busch et al., Int. Immunol. 2:443 (1990); Hill et al., J. Immunol. 147:189 (1991); del Guercio et al., J. Immunol. 154:685 (1995)), cell-free systems using detergent lysates (e.g., Cerundolo et al., J. Immunol. 21:2069 (1991)), immobilized purified MHC (e.g., Hill et al., J. Immunol. 152,2890 (1994); Marshall et al. al., J. Immunol. 152:4946 (1994)), ELISA systems (e.g., Reay, et al., EMBO J. 11:2829 (1992)), surface plasmon resonance (e.g., Khilko, et al., J. Biol. Chem. 268:15425 (1993)); high flux soluble phase (Hammer, et al., J. Exp. Med. 180:2353 (1994)), and measurements of class I MHC stabilization or assembly (e.g., Ljunggren, et al., Nature 346:476 (1990); Schumacher, et al., Cell 62:563 (1990); Townsend, et al., Cell 62:285 (1990); Parker, et al. The antibody can also be determined using other assay systems, including those using the antibody (e.g., IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IL-13, IL-14, IL-15, IL-16, IL-17, IL-18, IL-19,

[0089] Cap: As used herein, the term "cap" typically refers to a structure comprising or consisting essentially of a nucleoside-5'-triphosphate attached to the 5' end of an uncapped RNA (e.g., an uncapped RNA having a 5'-diphosphate). In some embodiments, the cap is or includes a guanine nucleotide. In some embodiments, the cap is or includes a naturally occurring RNA 5' cap, including, but not limited to, a 7-methylguanosine cap having the structure denoted "m7G." In some embodiments, the cap is or includes a synthetic cap analogue that resembles the RNA cap structure and has the ability to stabilize RNA when attached thereto (including, but not limited to, anti-reverse cap analogues (ARCAs) known in the art). Those of skill in the art will understand that methods for attaching a cap to the 5' end of an RNA are known to those of skill in the art. For example, in some embodiments, capped RNA can be obtained by in vitro capping of RNA bearing a 5' triphosphate group or RNA bearing a 5' diphosphate group with a capping enzyme system (e.g., including but not limited to, the vaccinia capping enzyme system or the Saccharomyces cerevisiae capping enzyme system). Alternatively, capped RNA can be obtained by in vitro transcription (IVT) of a single-stranded DNA template in the presence of a dinucleotide or trinucleotide cap analog.

[0090] Cell-mediated immunity: "Cell-mediated immunity," "cellular immunity," "cellular immune response," or similar terms are meant to include a cellular response to cells characterized by the expression of antigens, particularly cells characterized by the presentation of antigens with class I or class II MHC. The cellular response involves immune effector cells, particularly T cells or T lymphocytes, which act as either "helpers" or "killers." Helper T cells (CD4 + T cells (also called CD4 T cells) play a central role by regulating the immune response and are known as killer cells (cytotoxic T cells, cytolytic T cells, CD8+ T cells, also called CD8 T cells, or CTLs, kill diseased cells, such as virus-infected cells, and prevent the production of further diseased cells.

[0091] Co-administration: As used herein, the term "co-administration" refers to the use of a pharmaceutical composition (e.g., an immunogenic composition, e.g., a vaccine) described herein and an additional therapeutic agent. The combined use of a pharmaceutical composition (e.g., an immunogenic composition, e.g., a vaccine) described herein and an additional therapeutic agent may be performed simultaneously or separately (e.g., sequentially in any order). In some embodiments, a pharmaceutical composition (e.g., an immunogenic composition, e.g., a vaccine) described herein and an additional therapeutic agent may be combined in one pharmaceutically acceptable carrier, or they may be placed in separate carriers and delivered to a target cell or administered to a subject at different times. Each of these situations is intended to fall within the meaning of "co-administration" or "combination," provided that the pharmaceutical composition (e.g., an immunogenic composition, e.g., a vaccine) described herein and the additional therapeutic agent are delivered or administered sufficiently close in time that there is at least some temporal overlap in the biological effect(s) produced by each on the target cell or the subject being treated.

[0092] Codon optimization: As used herein, the term "codon optimization" refers to the modification of codons in the coding region of a nucleic acid molecule to reflect the typical codon usage of a host organism, preferably without modifying the amino acid sequence encoded by the nucleic acid molecule. Within the context of the present disclosure, in some embodiments, the coding region is codon-optimized for optimal expression in a subject treated using an RNA molecule described herein. In some embodiments, codon optimization can be performed such that, in place of a "rare codon," a codon for which a frequently occurring tRNA is available is inserted. In some embodiments, codon optimization can include increasing the guanosine / cytosine (G / C) content of the coding region of an RNA described herein compared to the G / C content of the corresponding coding sequence of a wild-type RNA, and the amino acid sequence encoded by the RNA is preferably unmodified compared to the amino acid sequence.

[0093] Combination therapy: As used herein, the term "combination therapy" refers to a situation in which a subject is exposed to two or more treatment regimens (e.g., two or more therapeutic agents) simultaneously. In some embodiments, the two or more regimens may be administered simultaneously. In some embodiments, the regimens may be administered sequentially (e.g., all "doses" of a first regimen are administered before any doses of a second regimen are administered). In some embodiments, the agents are administered in overlapping dosing regimens. In some embodiments, "administration" of a combination therapy may include administering one or more agent(s) or modality(s) in combination to a subject receiving other agent(s) or modality(s). For clarity, combination therapy does not require that the individual agents be administered together in a single composition (or even necessarily simultaneously), although in some embodiments, two or more agents or active portions thereof may be administered together in a combined composition.

[0094] Comparable: As used herein, the term "equivalent" refers to two or more agents, entities, circumstances, sets of conditions, etc. that may not be identical to one another, but that are sufficiently similar to permit a comparison between them where one of skill in the art would understand that conclusions can be reasonably drawn based on observed differences or similarities. In some embodiments, comparable sets of conditions, circumstances, individuals, or populations are characterized by multiple substantially identical characteristics and one or a few different characteristics. One of skill in the art will understand the degree of identity required to be considered comparable in any given situation for two or more such agents, entities, circumstances, sets of conditions, etc., in context. For example, one of skill in the art will understand that sets of circumstances, individuals, or populations are comparable to one another when they are characterized by a sufficient number and type of substantially identical characteristics to warrant a reasonable conclusion that differences in results obtained or phenomena observed under or with different sets of circumstances, individuals, or populations are caused by, or indicate, variations in those characteristics.

[0095] Corresponding to: As used herein, the term "corresponding to" refers to a relationship between two or more entities. For example, the term "corresponding to" can be used to indicate the location / identity of a structural element in a compound or composition relative to another compound or composition (e.g., an appropriate reference compound or composition). For example, in some embodiments, a monomer residue within a polymer (e.g., an amino acid residue within a polypeptide, or a nucleic acid residue within a polynucleotide) can be identified as "corresponding to" a residue in an appropriate reference polymer. For example, one of ordinary skill in the art will understand that, for simplicity's sake, residues within a polypeptide are often designated based on the relevant reference polypeptide using a standard numbering system, so that an amino acid "corresponding to" a residue at position 190, for example, corresponds to the residue found at 190 in the reference polypeptide, rather than necessarily being the actual 190th amino acid in a particular amino acid chain; one of ordinary skill in the art will readily understand how to identify a "corresponding" amino acid. For example, one of skill in the art will recognize various sequence alignment strategies, including, for example, software programs such as BLAST, CS-BLAST, CUSASW++, DIAMOND, FASTA, GGSEARCH / GLSEARCH, Genoogle, HMMER, HHpred / HHsearch, IDF, Infernal, KLAST, USEARCH, Parasail, PSI-BLAST, PSI-Search, ScalaBLAST, Sequilab, SAM, SSEARCH, SWAPHI, SWAPHI-LS, SWIMM, or SWIPE, that can be utilized to identify "corresponding" residues within polypeptides and / or nucleic acids in accordance with the present disclosure. One of skill in the art will also recognize that, in some cases, the term "corresponding to" can be used to describe an event or entity that shares relevant similarity with another event or entity (e.g., a suitable reference event or entity).As just one example, a gene or protein in one organism may, in some embodiments, be described as "corresponding to" a gene or protein from another organism to indicate that they play a similar role or perform a similar function, and / or exhibit a particular degree of sequence identity or homology, or share certain characteristic sequence elements.

[0096] Derived: In the context of an amino acid sequence (peptide or polypeptide) "derived from" a specified amino acid sequence (peptide or polypeptide), refers to a structural analog of the specified amino acid sequence. In some embodiments, an amino acid sequence derived from a particular amino acid sequence has an amino acid sequence that is identical, essentially identical, or homologous to the particular sequence or a fragment thereof. An amino acid sequence derived from a particular amino acid sequence may be a variant of the particular sequence or a fragment thereof. For example, it will be understood by those skilled in the art that antigens suitable for use herein can be modified such that the sequence varies from the naturally occurring sequence or the native sequence from which they are derived while retaining the desired activity of the native sequence.

[0097] Designed: As used herein, the term "designed" refers to (i) an agent whose structure is selected or chosen by the hand of man; (ii) an agent produced by a process requiring human intervention; and / or (iii) an agent that differs from natural substances and other known agents.

[0098] Dosing regimen: Those skilled in the art will understand that the term "dosing regimen" can be used to refer to a set of unit doses (typically more than one) administered individually to a subject, typically separated by time. In some embodiments, a given therapeutic agent has a recommended dosing regimen, which can include one or more doses. In some embodiments, a dosing regimen includes multiple doses, each spaced in time from the other doses. In some embodiments, the individual doses are separated from each other by equidistant periods of time. In some embodiments, a dosing regimen includes multiple doses, and the individual doses are separated by two different periods of time. In some embodiments, all doses within a dosing regimen are of the same unit dosage amount. In some embodiments, different doses within a dosing regimen are of different amounts. In some embodiments, a dosing regimen includes a first dose at a first dosage amount, followed by one or more additional doses at a second dosage amount different from the first dosage amount. In some embodiments, the dosing regimen comprises a first dose at a first dosage amount, followed by one or more additional doses at a second dosage amount that is the same as the first dosage amount. In some embodiments, the dosing regimen correlates with a desired or beneficial outcome when administered within a relevant population (i.e., is a therapeutic dosing regimen).

[0099] Encode: As used herein, the terms "encode" or "encoding" refer to the sequence information of a first molecule that directs the production of a second molecule having a defined sequence of nucleotides (e.g., mRNA) or a defined sequence of amino acids. For example, a DNA molecule can encode an RNA molecule (e.g., by a transcription process involving a DNA-dependent RNA polymerase enzyme). An RNA molecule can encode a polypeptide (e.g., by a translation process). Thus, a gene, cDNA, or RNA molecule (e.g., mRNA) encodes a polypeptide if transcription and translation of the mRNA corresponding to the gene produces the polypeptide in a cell or other biological system. In some embodiments, the coding region of an RNA molecule encoding a target antigen refers to the coding strand, the nucleotide sequence of which is identical to the mRNA sequence of such target antigen. In some embodiments, the coding region of an RNA molecule encoding a target antigen refers to the non-coding strand of such target antigen, which can be used as a template for transcription of the gene or cDNA.

[0100] Engineered: Generally, the term "engineered" refers to an aspect that has been manipulated by the hand of man. For example, a polynucleotide is considered to be "engineered" when it is manipulated by the hand of man so that two or more sequences that are not naturally linked together in that order are directly linked to each other in the engineered polynucleotide, and / or when certain residues within the polynucleotide are caused through the action of man to be linked to entities or moieties that are not naturally occurring and / or not naturally linked.

[0101] Epitope: As used herein, the term "epitope" refers to a portion that is specifically recognized by an immunoglobulin (e.g., an antibody or receptor) binding entity. For example, an epitope can be recognized by a T cell, a B cell, or an antibody. In some embodiments, an epitope is composed of multiple chemical atoms or groups on an antigen. In some embodiments, such chemical atoms or groups are surface-exposed when the antigen adopts a related three-dimensional conformation. In some embodiments, such chemical atoms or groups are physically close to each other in space when the antigen adopts such a conformation. In some embodiments, at least some such chemical atoms or groups are physically separated from each other when the antigen adopts an alternative conformation (e.g., is linearized). Thus, in some embodiments, an epitope of an antigen can comprise continuous or discontinuous fragments of the antigen. In some embodiments, an epitope is or comprises a T cell epitope. In some embodiments, an epitope can have a length of about 5 to about 30 amino acids, or about 10 to about 25 amino acids, or about 5 to about 15 amino acids, or about 5 to 12 amino acids, or about 6 to about 9 amino acids.

[0102] Expression: As used herein, the term "expression" of a nucleic acid sequence refers to the production of a gene product from the nucleic acid sequence. In some embodiments, the gene product can be a transcription product. In some embodiments, the gene product can be a polypeptide. In some embodiments, expression of a nucleic acid sequence involves one or more of: (1) generation of an RNA template from the DNA sequence (e.g., by transcription); (2) processing of the RNA transcript (e.g., by splicing, editing, etc.); (3) translation of the RNA into a polypeptide or protein; and / or (4) post-translational modification of the polypeptide or protein.

[0103] Five prime untranslated region: As used herein, the term "five prime untranslated region" or "5'UTR" refers to the sequence of an mRNA molecule between the transcription start site and the start codon of the coding region of the RNA. In some embodiments, "5'UTR" refers to the sequence of an mRNA molecule that begins at the transcription start site and ends one nucleotide (nt) before the start codon (usually AUG) of the coding region of the RNA molecule, e.g., in its natural context.

[0104] Fragment: As used herein in the context of a nucleic acid sequence (e.g., an RNA sequence) or an amino acid sequence, the term "fragment" typically refers to a fragment of a reference sequence. In some embodiments, the reference sequence is, for example, the full-length sequence of a nucleic acid sequence or an amino acid sequence. Thus, a fragment typically refers to a sequence that is identical to a corresponding stretch in the reference sequence. In some embodiments, a fragment comprises a contiguous stretch of nucleotides or amino acid residues that corresponds to at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 95% of the full length of the reference sequence from which the fragment is derived. In some embodiments, the term "fragment," with reference to an amino acid sequence (peptide or polypeptide), relates to a portion of the amino acid sequence, e.g., a sequence that represents an amino acid sequence truncated at the N-terminus and / or C-terminus. In some embodiments, a fragment of an amino acid sequence comprises at least 6, particularly at least 8, at least 12, at least 15, at least 20, at least 30, at least 50, or at least 100 contiguous amino acids from the amino acid sequence.

[0105] Homology: As used herein, the term "homology" or "homolog" refers to the overall relatedness between polynucleotide molecules (e.g., DNA molecules and / or RNA molecules) and / or polypeptide molecules. In some embodiments, polynucleotide molecules (e.g., DNA molecules and / or RNA molecules) and / or polypeptide molecules are considered to be "homologous" to one another if their sequences are at least 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% identical. In some embodiments, polynucleotide molecules (e.g., DNA molecules and / or RNA molecules) and / or polypeptide molecules are considered to be "homologous" to one another if their sequences are at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% similar (e.g., contain residues with related chemical properties at corresponding positions). For example, as is well known to those of skill in the art, certain amino acids are typically classified as similar to one another as "hydrophobic" or "hydrophilic" amino acids and / or as having "polar" or "nonpolar" side chains. Substitution of one amino acid for another amino acid of the same type can often be considered a "homologous" substitution.

[0106] Humoral immunity: As used herein, the term "humoral immunity" or "humoral immune response" refers to antibody production and its associated accessory processes, including Th2 activation and cytokine production, germinal center formation and isotype switching, affinity maturation, and memory cell generation. It also refers to antibody effector functions, including pathogen neutralization, classical complement activation, and opsonization to facilitate phagocytosis and pathogen elimination.

[0107] Identity: As used herein, the term "identity" refers to the overall relatedness between polynucleotide molecules (e.g., DNA molecules and / or RNA molecules) and / or polypeptide molecules. In some embodiments, polynucleotide molecules (e.g., DNA molecules and / or RNA molecules) and / or polypeptide molecules are considered to be "substantially identical" to one another if their sequences are at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical. For example, calculation of the percent identity of two nucleic acid or polypeptide sequences can be performed by aligning the two sequences for optimal comparison purposes (e.g., gaps can be introduced into one or both of the first and second sequences for optimal alignment, and non-identical sequences can be disregarded for comparison purposes). In certain embodiments, the length of an aligned sequence for comparison purposes is at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or substantially 100% of the length of the reference sequence. The nucleotides at corresponding positions are then compared. If a position in the first sequence is occupied by the same residue (e.g., nucleotide or amino acid) as the corresponding position in the second sequence, the molecules are identical at that position. The percent identity between two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps and the length of each gap that needs to be introduced for optimal alignment of the two sequences. Sequence comparison and determination of percent identity between two sequences can be accomplished using a mathematical algorithm. For example, the algorithm of Meyers and Miller, 1989, incorporated into the ALIGN program (version 2.0), can be used to determine the percent identity between two nucleotide sequences.In some exemplary embodiments, nucleic acid sequence comparisons generated with the ALIGN program use a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4. The percent identity between two nucleotide sequences can alternatively be determined using the GAP program in the GCG software package using the NWSgapdna.CMP matrix.

[0108] Immunologically equivalent: The term "immunologically equivalent" means that immunologically equivalent molecules, such as immunologically equivalent amino acid sequences, exhibit the same or essentially the same immunological properties and / or exert the same or essentially the same immunological effect, e.g., with respect to the type of immunological effect. In the context of the present disclosure, in some embodiments, the term "immunologically equivalent" is used with reference to the immunological effect or properties of an antigen or antigen variant used for immunization. For example, an amino acid sequence is immunologically equivalent to a reference amino acid sequence if, when exposed to a subject's immune system, the amino acid sequence elicits an immune response with specificity that reacts with the reference amino acid sequence.

[0109] In one embodiment, the antigen receptor is an antibody or B cell receptor that binds to an epitope of the antigen. In one embodiment, the antibody or B cell receptor binds to a natural epitope of the antigen.

[0110] Increase, induce, or reduce: As used herein, these terms, or grammatically equivalent comparative terms, refer to values ​​that are relative to an equivalent reference measurement. For example, in some embodiments, a value achieved by a provided pharmaceutical composition (e.g., an immunogenic composition, e.g., a vaccine) may be "increased" compared to that obtained by an equivalent reference pharmaceutical composition (e.g., an immunogenic composition, e.g., a vaccine). Alternatively or additionally, in some embodiments, a value achieved in a subject may be "increased" compared to that obtained in the same subject under different conditions (e.g., before and after an event), or in the presence or absence of an event, such as administration of a pharmaceutical composition (e.g., an immunogenic composition, e.g., a vaccine) as described herein, or in a different comparable subject (e.g., in a comparable subject different from the subject of interest, in the absence of administration of a pharmaceutical composition (e.g., an immunogenic composition, e.g., a vaccine) as described herein, upon prior exposure to a condition). In some embodiments, comparative terms refer to a statistically relevant difference (e.g., one of sufficient prevalence and / or magnitude to achieve statistical relevance). Those skilled in the art will recognize or be able to readily determine the degree and / or prevalence of difference necessary or sufficient to achieve such statistical significance in a given context. In some embodiments, "reduced" or equivalent terms refer to a reduction in the level of an evaluated value by at least 5%, at least 10%, at least 20%, at least 50%, at least 75%, or more, compared to an equivalent reference. In some embodiments, the term "reduced" or equivalent terms refer to complete or essentially complete inhibition, i.e., a reduction to zero, or a reduction to essentially zero. In some embodiments, the term "increased" or "induced" refers to an increase in the level of an evaluated value by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 80%, at least 100%, at least 200%, at least 500%, or more, compared to an equivalent reference.

[0111] Ionizable: The term "ionizable" refers to a compound, group, or atom that is charged at a certain pH. In the context of ionizable amino lipids, such lipids or their functional groups or atoms have a positive charge at a certain pH. In some embodiments, ionizable amino lipids are positively charged at acidic pH. In some embodiments, ionizable amino lipids are primarily neutral at physiological pH values, e.g., in some embodiments, about 7.0 to 7.4, but become positively charged at lower pH values. In some embodiments, ionizable amino lipids may have a pKa in the range of about 5 to about 7.

[0112] Isolated: The term "isolated" means altered or removed from the natural state. For example, a nucleic acid or peptide that is naturally present in a living animal is not "isolated," but the same nucleic acid or peptide that has been partially or completely separated from the coexisting materials of its natural state is "isolated." An isolated nucleic acid or protein can exist in a substantially purified form, or can exist in a non-native environment, such as, for example, a host cell.

[0113] Lipid: As used herein, the terms "lipid" and "lipid-like material" are broadly defined as molecules that contain one or more hydrophobic moieties or groups, and optionally also one or more hydrophilic moieties or groups. Molecules that contain hydrophobic and hydrophilic moieties are also typically referred to as amphiphiles.

[0114] RNA lipid nanoparticles: As used herein, the term "RNA lipid nanoparticles" refers to nanoparticles comprising at least one lipid and RNA molecule(s). In some embodiments, the RNA lipid nanoparticles comprise at least one ionizable amino lipid. In some embodiments, the RNA lipid nanoparticles comprise at least one ionizable amino lipid, at least one helper lipid, and at least one polymer-conjugated lipid (e.g., a PEG-conjugated lipid). In various embodiments, the RNA lipid nanoparticles described herein can have an average size (e.g., Z-average) of about 100 nm to 1000 nm, or about 200 nm to 900 nm, or about 200 nm to 800 nm, or about 250 nm to about 700 nm. In some embodiments of the present disclosure, the RNA-lipid nanoparticles can have a particle size (e.g., Z-average) of about 30 nm to about 200 nm, or about 30 nm to about 150 nm, about 40 nm to about 150 nm, about 50 nm to about 150 nm, about 60 nm to about 130 nm, about 70 nm to about 110 nm, about 70 nm to about 100 nm, about 80 nm to about 100 nm, about 90 nm to about 100 nm, about 70 nm to about 90 nm, about 80 nm to about 90 nm, or about 70 nm to about 80 nm. In some embodiments, the average size of the lipid nanoparticles is determined by measuring particle size. In some embodiments, the RNA-lipid nanoparticles can be prepared by mixing lipids with the RNA molecules described herein.

[0115] Lipidoid: As used herein, "lipidoid" refers to a lipid-like molecule. In some embodiments, lipids are amphipathic molecules that have one or more lipid-like physical properties. In the context of this disclosure, the term lipid is considered to encompass lipidoids.

[0116] Nanoparticles: As used herein, the term "nanoparticles" refers to particles having an average size suitable for parenteral administration. In some embodiments, nanoparticles have a longest dimension (e.g., diameter) of less than 1,000 nanometers (nm). In some embodiments, nanoparticles can be characterized by a longest dimension (e.g., diameter) of less than 300 nm. In some embodiments, nanoparticles can be characterized by a longest dimension (e.g., diameter) of less than 100 nm. In many embodiments, nanoparticles can be characterized by a longest dimension of about 1 nm to about 100 nm, or about 1 μm to about 500 nm, or about 1 nm to 1,000 nm. In many embodiments, a population of nanoparticles is characterized by an average size (e.g., longest dimension) of less than about 1,000 nm, about 500 nm, about 100 nm, about 50 nm, about 40 nm, about 30 nm, about 20 nm, or about 10 nm, and often greater than about 1 nm. In many embodiments, nanoparticles can be substantially spherical, such that their longest dimension can be their diameter. In some embodiments, the nanoparticles have a diameter of less than 100 nm, as defined by the National Institutes of Health.

[0117] Naturally occurring: As used herein, the term "naturally occurring" refers to an entity that can be found in nature. For example, a peptide or nucleic acid that is present in an organism (including viruses), can be isolated from a natural source, and has not been intentionally modified by man in a laboratory is naturally occurring.

[0118] Neutralization: As used herein, the term "neutralization" refers to an event in which a binding agent, such as an antibody, binds to a biologically active site of a virus, such as a receptor-binding protein, thereby inhibiting parasitic infection of a cell. In some embodiments, the term "neutralization" refers to an event in which the ability of the binding agent to infect a cell is eliminated or significantly reduced.

[0119] Nucleic acid particles: "Nucleic acid particles" can be used to deliver nucleic acids to a desired target site (e.g., a cell, tissue, organ, etc.). Nucleic acid particles can include at least one cationic lipid or cationically ionizable lipid or lipid-like material, at least one cationic polymer such as protamine, or a mixture thereof, and nucleic acid. In some embodiments, the nucleic acid particles are lipid nanoparticles. In some embodiments, the nucleic acid particles are lipoplex particles.

[0120] Nucleic Acid / Polynucleotide: As used herein, the term "nucleic acid" refers to a polymer of at least 10 or more nucleotides. In some embodiments, a nucleic acid is or comprises DNA. In some embodiments, a nucleic acid is or comprises RNA. In some embodiments, a nucleic acid is or comprises peptide nucleic acid (PNA). In some embodiments, a nucleic acid is or comprises single-stranded nucleic acid. In some embodiments, a nucleic acid is or comprises double-stranded nucleic acid. In some embodiments, a nucleic acid comprises both single-stranded and double-stranded fragments. In some embodiments, a nucleic acid comprises a backbone comprising one or more phosphodiester bonds. In some embodiments, a nucleic acid comprises a backbone comprising both phosphodiester and non-phosphodiester bonds. For example, in some embodiments, a nucleic acid may comprise a backbone comprising one or more phosphorothioate or 5'-N-phosphoramidite bonds and / or one or more peptide bonds, e.g., "peptide nucleic acids." In some embodiments, a nucleic acid comprises one or more, or all, naturally occurring residues (e.g., adenine, cytosine, deoxyadenosine, deoxycytidine, deoxyguanosine, deoxythymidine, guanine, thymine, uracil). In some embodiments, a nucleic acid comprises one or more, or all, non-naturally occurring residues. In some embodiments, the non-natural residue comprises a nucleoside analog (e.g., 2-aminoadenosine, 2-thiothymidine, inosine, pyrrolo-pyrimidine, 3-methyladenosine, 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, 6-O-methylguanine, 2-thiocytidine, methylated bases, intercalating bases, and combinations thereof).In some embodiments, the non-natural residue comprises one or more modified sugars (e.g., 2'-fluororibose, ribose, 2'-deoxyribose, arabinose, and hexose) compared to that of the natural residue. In some embodiments, the nucleic acid has a nucleotide sequence that encodes a functional gene product, such as an RNA or a polypeptide. In some embodiments, the nucleic acid has a nucleotide sequence that includes one or more introns. In some embodiments, nucleic acids can be prepared by isolation from natural sources, enzymatic synthesis (e.g., polymerization based on a complementary template in vivo or in vitro, replication in a recombinant cell or system, or chemical synthesis. In some embodiments, nucleic acids are at least 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 20, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 600, 700, 800, 900, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, 5000, 5500, 6000, 6500, 7000, 7500, 8000, 8500, 9000, 9500, 10,000, 10,500, 11,000, 11,500, 12,000, 12 1,500, 13,000, 13,500, 14,000, 14,500, 15,000, 15,500, 16,000, 16,500, 17,000, 17,500, 18,000, 18,500, 19,000, 19,500, or 20,000 or more residues or nucleotides in length.

[0121] Nucleotide: As used herein, the term "nucleotide" refers to its art-recognized meaning. When a number of nucleotides is used, for example, as an indicator of the size of a polynucleotide, a particular number of nucleotides refers to the number of nucleotides on a single strand, for example, a polynucleotide.

[0122] Patient: As used herein, the term "patient" refers to any organism suffering from or at risk of a disease or disorder or condition. Typical patients include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, and / or humans). In some embodiments, the patient is human. In some embodiments, the patient is suffering from or susceptible to one or more diseases or disorders or conditions. In some embodiments, the patient exhibits one or more symptoms of a disease or disorder or condition. In some embodiments, the patient has been diagnosed with one or more diseases or disorders or conditions. In some embodiments, the disease or disorder or condition amenable to the provided techniques is or includes an HSV infection. In some embodiments, the patient is undergoing or has undergone a particular therapy to diagnose and / or treat the disease, disorder, or condition. In some embodiments, the patient is suffering from or susceptible to an HSV infection.

[0123] PEG-conjugated lipid: The term "PEG-conjugated lipid" refers to a molecule comprising a lipid moiety and a polyethylene glycol moiety.

[0124] Pharmaceutical composition: As used herein, the term "pharmaceutical composition" refers to an active agent formulated with one or more pharmaceutically acceptable carriers. In some embodiments, the active agent is present in a unit dosage suitable for administration in a treatment regimen that exhibits a statistically significant likelihood of achieving a predetermined therapeutic effect when administered to a relevant population. In some embodiments, the pharmaceutical composition may be specifically formulated for parenteral administration, e.g., by subcutaneous, intramuscular, or intravenous injection, e.g., as a sterile solution or suspension formulation.

[0125] Pharmaceutically effective amount: The term "pharmaceutically effective amount" or "therapeutically effective amount" refers to an amount that achieves a desired response or desired effect, either alone or together with further doses. In the case of treatment of a particular disease, the desired response in some embodiments relates to inhibition of the course of the disease. In some embodiments, such inhibition may include slowing and / or halting or reversing the course of the disease. In some embodiments, the desired response in the treatment of a disease may be or may include delaying or preventing the onset of the disease or condition. The effective amount of a pharmaceutical composition (e.g., an immunogenic composition, e.g., a vaccine) described herein will depend on individual patient parameters, including, for example, the disease or condition to be treated, the severity of such disease or condition, e.g., age, physiological condition, size, and weight, the duration of treatment, the type of concomitant therapy (if any), the particular route of administration, and similar factors. Thus, the dose of a pharmaceutical composition (e.g., an immunogenic composition, e.g., a vaccine) described herein may depend on various such parameters. If the patient responds inadequately to the initial dose, a higher dose (or an effectively higher dose achieved by a different, more localized route of administration) may be used.

[0126] Poly(A) sequence: As used herein, the term "poly(A) sequence" or "poly(A tail)" typically refers to an uninterrupted or interrupted sequence of adenylate residues located at the 3' end of an RNA molecule. Poly(A) sequences are known to those skilled in the art and may follow the 3'UTR in the RNAs described herein. Uninterrupted poly(A) sequences are characterized by consecutive adenylate residues. Uninterrupted poly(A) sequences are typical in nature. The RNAs disclosed herein may have a poly(A) sequence attached to the free 3' end of the RNA by a non-template-dependent RNA polymerase after transcription, or a poly(A) sequence encoded by DNA and transcribed by a template-dependent RNA polymerase.

[0127] Polypeptide: As used herein, the term "polypeptide" refers to a polymeric chain of amino acids. In some embodiments, a polypeptide has a naturally occurring amino acid sequence. In some embodiments, a polypeptide has a non-naturally occurring amino acid sequence. In some embodiments, a polypeptide has an engineered amino acid sequence, in that it has been designed and / or produced through the action of man. In some embodiments, a polypeptide may comprise or consist of natural amino acids, unnatural amino acids, or both. In some embodiments, a polypeptide may comprise or consist of only natural amino acids or only unnatural amino acids. In some embodiments, a polypeptide may comprise D-amino acids, L-amino acids, or both. In some embodiments, a polypeptide may comprise only D-amino acids. In some embodiments, a polypeptide may comprise only L-amino acids. In some embodiments, a polypeptide may comprise one or more pendant groups or other modifications, e.g., modifications of or attachment to one or more amino acid side chains, at the N-terminus of the polypeptide, the C-terminus of the polypeptide, or any combination thereof. In some embodiments, such pendant groups or modifications include acetylation, amidation, lipidation, methylation, pegylation, etc., e.g., combinations thereof. In some embodiments, a polypeptide may be cyclic and / or include a cyclic portion. In some embodiments, a polypeptide is not cyclic and / or does not include a cyclic portion. In some embodiments, a polypeptide is linear. In some embodiments, a polypeptide may be or include a stapled polypeptide. In some embodiments, the term "polypeptide" may be appended to the name of a reference polypeptide, activity, or structure, and in such cases, it is used herein to refer to polypeptides that share a related activity or structure and can therefore be considered members of the same class or family of polypeptides.For each such class, the present specification provides, and / or one of skill in the art will recognize, exemplary polypeptides within the class whose amino acid sequence and / or function are known. In some embodiments, such exemplary polypeptides are reference polypeptides of a class or family of polypeptides. In some embodiments, members of a polypeptide class or family exhibit significant sequence homology or identity with the reference polypeptide of the class (and, in some embodiments, with all polypeptides in the class), share common sequence motifs (e.g., characteristic sequence elements), and / or share a common activity (in some embodiments, at a similar level or within a specified range) with the reference polypeptide of the class (and, in some embodiments, with all polypeptides in the class). For example, in some embodiments, a member polypeptide exhibits an overall degree of sequence homology or identity with a reference polypeptide of at least about 30-40%, and often greater than about 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more, and / or contains at least one region (e.g., a conserved region, which in some embodiments may be or may include a distinctive sequence element) that exhibits very high sequence identity, often greater than 90%, or even greater than 95%, 96%, 97%, 98%, or 99%. Such a conserved region typically encompasses at least 3-4, and often up to 20 or more, amino acids; in some embodiments, the conserved region encompasses at least one stretch of at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or more contiguous amino acids. In some embodiments, the related polypeptide may comprise or consist of a fragment of a parent polypeptide.

[0128] Prevention: As used herein, the terms "prevent" or "prevention," when used in reference to the occurrence of a disease, disorder, and / or condition, refers to a reduction in the risk of the disease, disorder, and / or condition occurring and / or a delay in the onset of one or more characteristics or symptoms of the disease, disorder, or condition. Prevention may be considered complete if the onset of the disease, disorder, or condition has been delayed for a predefined period of time.

[0129] Recombinant: The term "recombinant," in the context of the present disclosure, means "produced by genetic engineering." In some embodiments, a "recombinant" entity, such as a recombinant nucleic acid, in the context of the present disclosure is not naturally occurring.

[0130] Reference: As used herein, the term "reference" describes a standard or control against which a comparison is made. For example, in some embodiments, an agent, animal, individual, population, sample, sequence, or value of interest is compared to a reference or control agent, animal, individual, population, sample, sequence, or value. In some embodiments, the reference or control is tested and / or determined substantially contemporaneously with the test or determination of interest. In some embodiments, the reference or control is a historical reference or control, optionally embodied in a tangible medium. Typically, as will be understood by those of skill in the art, a reference or control is determined or characterized under conditions or circumstances comparable to those being evaluated. Those of skill in the art will understand when there is sufficient similarity to justify reliance on and / or comparison to a particular reference or control considered.

[0131] Ribonucleic acid (RNA): As used herein, the term "RNA" refers to a polymer of ribonucleotides. In some embodiments, the RNA is single-stranded. In some embodiments, the RNA is double-stranded. In some embodiments, the RNA includes both single-stranded and double-stranded fragments. In some embodiments, the RNA can include a backbone structure as described in the definition of "nucleic acid / polynucleotide" above. The RNA can be a regulatory RNA (e.g., siRNA, microRNA, etc.) or a messenger RNA (mRNA). In some embodiments, the RNA is an mRNA. In some embodiments, the RNA is an mRNA, and the RNA typically includes a poly(A) region at its 3' end. In some embodiments, the RNA is an mRNA, and the RNA typically includes an art-recognized cap structure at its 5' end, for example, for recognition and attachment of the mRNA to a ribosome to initiate translation. In some embodiments, the RNA is synthetic RNA. Synthetic RNA includes RNA synthesized in vitro (e.g., by enzymatic and / or chemical synthesis).

[0132] Ribonucleotide: As used herein, the term "ribonucleotide" encompasses unmodified ribonucleotides and modified ribonucleotides. For example, unmodified ribonucleotides include the purine bases adenine (A) and guanine (G) and the pyrimidine bases cytosine (C) and uracil (U). Modified ribonucleotides can include one or more modifications, including, but not limited to, (a) terminal modifications, such as 5'-terminal modifications (e.g., phosphorylation, dephosphorylation, conjugation, inverted linkages, etc.), 3'-terminal modifications (e.g., conjugation, inverted linkages, etc.), (b) base modifications, such as replacement with a modified base, a stabilized base, a destabilized base, or a base that base pairs with an expanded repertoire of partners, or a conjugated base, (c) sugar modifications (e.g., at the 2' or 4' position) or sugar replacement, and (d) internucleoside linkage modifications, including modification or replacement of a phosphodiester bond. The term "ribonucleotide" also encompasses ribonucleotide triphosphates, including modified and unmodified ribonucleotide triphosphates.

[0133] Risk: As understood from the context, "risk" of a disease, disorder, and / or condition refers to the likelihood that a particular individual will develop the disease, disorder, and / or condition. In some embodiments, risk is expressed as a percentage. In some embodiments, risk is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, up to 100%. In some embodiments, risk is expressed as risk compared to the risk associated with a reference sample or group of reference samples. In some embodiments, the reference sample or group of reference samples has a known risk of the disease, disorder, condition, and / or event. In some embodiments, the reference sample or group of reference samples is from individuals comparable to the particular individual. In some embodiments, the relative risk is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more. In some embodiments, risk may reflect, for example, one or more genetic attributes that may (or may not) predispose an individual to developing a particular disease, disorder, and / or condition, hi some embodiments, risk may reflect one or more epigenetic events or attributes, and / or one or more lifestyle or environmental events or attributes.

[0134] RNA lipoplex particles: As used herein, the term "RNA lipoplex particles" refers to a complex comprising a liposome, particularly a cationic liposome, and an RNA molecule. Without wishing to be bound by any particular theory, electrostatic interactions between positively charged liposomes and negatively charged RNA result in the complexation and spontaneous formation of RNA lipoplex particles. In some embodiments, the positively charged liposome may comprise a cationic lipid, e.g., in some embodiments, DOTMA, and an additional lipid, e.g., in some embodiments, DOPE. In one embodiment, the RNA lipoplex particles are nanoparticles.

[0135] Selective or specific: The terms "selective" or "specific," as used herein with respect to an active agent, are understood by those skilled in the art to mean that the agent discriminates among potential target entities, conditions, or cells. For example, in some embodiments, an agent is said to "specifically" bind to a target if it preferentially binds to that target in the presence of one or more competing alternative targets. In many embodiments, the specific interaction depends on the presence of a particular structural feature of the target entity (e.g., an epitope, cleft, binding site). It should be understood that specificity need not be absolute. In some embodiments, specificity can be assessed relative to the specificity of the target binding moiety for one or more other potential target entities (e.g., a competitor). In some embodiments, specificity is assessed relative to the specificity of a reference specific binding moiety. In some embodiments, specificity is assessed relative to the specificity of a reference nonspecific binding moiety.

[0136] Stable: As used herein, the term "stable" in the context of the present disclosure refers to a pharmaceutical composition (e.g., an immunogenic composition, e.g., a vaccine) as a whole and / or its components that meet or exceed predetermined acceptance criteria. For example, in some embodiments, a stable pharmaceutical composition (e.g., an immunogenic composition, e.g., a vaccine) does not exhibit unacceptable levels of microbial growth and degradation or degradation of the active biomolecular component(s) is substantially absent or nonexistent. In some embodiments, a stable pharmaceutical composition (e.g., an immunogenic composition, e.g., a vaccine) refers to the integrity of the RNA molecules being maintained at least 90% or more. In some embodiments, a stable pharmaceutical composition (e.g., an immunogenic composition, e.g., a vaccine) refers to at least 90% or more (e.g., comprising at least 95%, at least 96%, at least 97%, or more) of the RNA molecules being maintained as encapsulated within the lipid nanoparticles. In some embodiments, a stable pharmaceutical composition (e.g., an immunogenic composition, e.g., a vaccine) refers to a formulation that remains capable of eliciting a desired immune response when administered to a subject. In some embodiments, a pharmaceutical composition (eg, an immunogenic composition, eg, a vaccine) remains stable under certain conditions for a certain period of time.

[0137] Subject: As used herein, the term "subject" refers to an organism administered with a composition described herein, for example, for experimental, diagnostic, prophylactic, and / or therapeutic purposes. Typical subjects include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, domestic pets, etc.) and humans. In some embodiments, the subject is a human subject. In some embodiments, the subject is afflicted with a disease, disorder, or condition (e.g., HSV infection). In some embodiments, the subject is susceptible to a disease, disorder, or condition (e.g., HSV infection). In some embodiments, the subject exhibits one or more symptoms or characteristics of a disease, disorder, or condition (e.g., HSV infection). In some embodiments, the subject exhibits one or more non-specific symptoms of a disease, disorder, or condition (e.g., HSV infection). In some embodiments, the subject does not exhibit any symptoms or characteristics of a disease, disorder, or condition (e.g., HSV infection). In some embodiments, the subject possesses one or more characteristics characteristic of susceptibility to or risk for a disease, disorder, or condition (e.g., HSV infection). In some embodiments, the subject is a patient. In some embodiments, the subject is an individual to whom and / or to whom a diagnosis and / or therapy is to be administered.

[0138] Suffering from: An individual "suffering from" a disease, disorder, and / or condition has and / or exhibits one or more symptoms of the disease, disorder, and / or condition.

[0139] Susceptible to: An individual "susceptible to" a disease, disorder, and / or condition is an individual who is at a higher risk than members of the general public of developing the disease, disorder, and / or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition may not have been diagnosed with the disease, disorder, and / or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition may exhibit symptoms of the disease, disorder, and / or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition may not exhibit symptoms of the disease, disorder, and / or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition develops the disease, disorder, and / or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition does not develop the disease, disorder, and / or condition.

[0140] Synthetic: As used herein, the term "synthetic" refers to an entity that is artificial, or created by human intervention, or that does not occur in nature, but rather results from synthesis. For example, in some embodiments, a synthetic nucleic acid or polynucleotide refers to a nucleic acid molecule that is chemically synthesized, e.g., in some embodiments, by solid-phase synthesis. In some embodiments, the term "synthetic" refers to an entity that is made outside of a biological cell. For example, in some embodiments, a synthetic nucleic acid or polynucleotide refers to a nucleic acid molecule (e.g., RNA) that is produced by in vitro transcription using a template.

[0141] Therapy: The term "therapy" refers to the administration or delivery of an agent or intervention that has a therapeutic effect and / or induces a desired biological and / or pharmacological effect (e.g., that has been demonstrated to be statistically likely to have such an effect when administered to a relevant population). In some embodiments, a therapeutic agent or therapy is any substance that can be used to alleviate, ameliorate, relieve, inhibit, prevent, delay onset, reduce severity, and / or reduce incidence of one or more symptoms or characteristics of a disease, disorder, and / or condition. In some embodiments, a therapeutic agent or therapy is a medical intervention (e.g., surgery, radiation, phototherapy) that can be performed to alleviate, alleviate, inhibit, present, delay onset, reduce severity, and / or reduce incidence of one or more symptoms or characteristics of a disease, disorder, and / or condition.

[0142] Three prime untranslated region: As used herein, the term "three prime untranslated region" or "3'UTR" refers to the sequence of an mRNA molecule that begins after the stop codon of the coding region of an open reading frame sequence. In some embodiments, the 3'UTR begins immediately after the stop codon of the coding region of an open reading frame sequence, e.g., in its natural context. In other embodiments, the 3'UTR does not begin immediately after the stop codon of the coding region of an open reading frame sequence, e.g., in its natural context.

[0143] Threshold level (e.g., acceptance criteria): As used herein, the term "threshold level" refers to a level used as a reference for obtaining information about and / or classifying the results of a measurement, e.g., the results of a measurement achieved in an assay. For example, in some embodiments, the threshold level refers to a value measured in an assay that defines a dividing line between two subsets of a population (e.g., batches that meet quality control standards versus batches that do not meet quality control standards). Thus, values ​​at or above the threshold level define one subset of a population, and values ​​below the threshold level define another subset of the population. The threshold level can be determined based on one or more control samples or across a population of control samples. The threshold level can be determined before, simultaneously with, or after the measurement of interest is performed. In some embodiments, the threshold level can be a range of values.

[0144] Treat: As used herein, the terms "treat," "treatment," or "treating" refer to any method used to partially or completely alleviate, ameliorate, relieve, inhibit, prevent, delay the onset of, reduce the severity of, and / or reduce the incidence of one or more symptoms or characteristics of a disease, disorder, and / or condition. Treatment may be administered to a subject who does not show signs of a disease, disorder, and / or condition. In some embodiments, treatment may be administered to a subject who shows only early signs of a disease, disorder, and / or condition, e.g., for the purpose of reducing the risk of developing pathologies associated with the disease, disorder, and / or condition. In some embodiments, treatment may be administered to a subject at a later stage of a disease, disorder, and / or condition.

[0145] Vaccination: As used herein, the term "vaccination" refers to the administration of a composition intended to generate an immune response, for example, to a disease-associated (e.g., pathogenic) agent. In some embodiments, vaccination can be administered before, during, and / or after exposure to the disease-associated agent, and in certain embodiments, before, during, and / or immediately after exposure to the agent. In some embodiments, vaccination involves multiple administrations of the vaccine composition, appropriately spaced in time. In some embodiments, vaccination generates an immune response against an infectious agent.

[0146] Vaccine: As used herein, the term "vaccine" refers to a composition that induces an immune response upon administration to a subject. In some embodiments, the induced immune response provides protective immunity.

[0147] Variant: As used herein, in the context of a molecule, e.g., a nucleic acid, protein, or small molecule, the term "variant" refers to a molecule that exhibits significant structural identity with a reference molecule but differs structurally from the reference molecule, e.g., in the presence or absence or level of one or more chemical moieties compared to the reference entity. In some embodiments, a variant also differs functionally from its reference molecule. Generally, whether a particular molecule is properly considered a "variant" of a reference molecule is based on the degree of structural identity with the reference molecule. As will be understood by those skilled in the art, any biological or chemical reference molecule possesses certain characteristic structural elements. A variant, by definition, is a distinct molecule that shares one or more such characteristic structural elements but differs in at least one aspect from the reference molecule. In some embodiments, a variant polypeptide or nucleic acid may differ from a reference polypeptide or nucleic acid as a result of one or more differences in amino acid or nucleotide sequence and / or one or more differences in chemical moieties (e.g., carbohydrate, lipid, phosphate group) that are covalent components of the polypeptide or nucleic acid (e.g., to which the polypeptide or nucleic acid backbone is attached). In some embodiments, the variant polypeptide or nucleic acid exhibits at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, or 99% overall sequence identity with the reference polypeptide or nucleic acid. In some embodiments, the variant polypeptide or nucleic acid does not share at least one characteristic sequence element with the reference polypeptide or nucleic acid. In some embodiments, the reference polypeptide or nucleic acid has one or more biological activities. In some embodiments, the variant polypeptide or nucleic acid shares one or more of the biological activities of the reference polypeptide or nucleic acid. In some embodiments, the variant polypeptide or nucleic acid lacks one or more of the biological activities of the reference polypeptide or nucleic acid. In some embodiments, the variant polypeptide or nucleic acid exhibits a reduced level of one or more biological activities compared to the reference polypeptide or nucleic acid.In some embodiments, a polypeptide or nucleic acid of interest is considered a "variant" of a reference polypeptide or nucleic acid if it has an amino acid or nucleotide sequence identical to that of the reference, but with a small number of sequence modifications at specific positions. Typically, less than about 20%, about 15%, about 10%, about 9%, about 8%, about 7%, about 6%, about 5%, about 4%, about 3%, or about 2% of the residues in the variant are substituted, inserted, or deleted compared to the reference. In some embodiments, a variant polypeptide or nucleic acid contains about 10, about 9, about 8, about 7, about 6, about 5, about 4, about 3, about 2, or about 1 substituted residue compared to the reference. In many cases, a variant polypeptide or nucleic acid contains a very small number (e.g., less than about 5, about 4, about 3, about 2, or about 1) of functional residues (i.e., residues involved in a specific biological activity) substituted, inserted, or deleted compared to the reference. In some embodiments, the variant polypeptide or nucleic acid contains no more than about 5, about 4, about 3, about 2, or about 1 additions or deletions, and in some embodiments no additions or deletions, compared to the reference. In some embodiments, the variant polypeptide or nucleic acid contains less than about 25, about 20, about 19, about 18, about 17, about 16, about 15, about 14, about 13, about 10, about 9, about 8, about 7, about 6, and typically less than about 5, about 4, about 3, or about 2 additions or deletions, compared to the reference. In some embodiments, the reference polypeptide or nucleic acid is one found in nature.

[0148] Vector: As used herein, refers to a nucleic acid molecule capable of transporting another nucleic acid linked to it. One type of vector is a "plasmid," which refers to a circular double-stranded DNA loop into which additional DNA segments can be ligated. Another type of vector is a viral vector, into which additional DNA segments can be ligated. Certain vectors are capable of autonomous replication in a host cell into which they are introduced (e.g., bacterial vectors having a bacterial origin of replication and episomal mammalian vectors). Other vectors (e.g., non-episomal mammalian vectors) can be integrated into the genome of a host cell upon introduction into the host cell, and thereby are replicated along with the host genome. Moreover, some vectors are capable of directing the expression of genes to which they are operably linked. Such vectors are referred to herein as "expression vectors." In some embodiments, known techniques can be used, for example, for recombinant DNA production or manipulation, 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 techniques and procedures described above can generally be carried out according to conventional methods well known in the art and as described in various general and more specific references cited and discussed throughout this specification. See, for example, Sambrook et al., Molecular Cloning: A Laboratory Manual (4th ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (2012)), which is incorporated herein by reference for any purpose.

[0149] All literature and similar materials cited in this application, including, but not limited to, patents, patent applications, articles, books, papers, and web pages, regardless of the format of such literature and similar materials, are expressly incorporated by reference in their entirety. In the event that one or more of the incorporated literature and similar materials, including, but not limited to, defined terms, term usage, described techniques, or the like, differs from or conflicts with this application, this application controls. The section headings used herein are for organizational purposes only and should not be construed as limiting the subject matter described in any way. [Mode for Carrying Out the Invention]

[0150] As discussed above, the present disclosure provides pharmaceutical compositions (e.g., immunogenic compositions, e.g., vaccines) and related technologies (e.g., methods) for delivering certain herpes simplex virus (HSV) antigen constructs (e.g., HSV-1 antigen constructs, HSV-2 antigen constructs, or combinations thereof) to a subject (e.g., a patient). In particular, the present disclosure provides HSV (e.g., HSV-1, HSV-2, or both) vaccine compositions and related technologies (e.g., methods).

[0151] The present disclosure provides, for example, polyribonucleotides encoding one or more HSV antigens. In some embodiments, such polyribonucleotides can be part of an RNA construct. In some embodiments, the polyribonucleotides or RNA constructs described herein can be part of a composition (e.g., a pharmaceutical composition, e.g., an immunogenic composition, e.g., a vaccine).

[0152] In some embodiments, the technology provided herein is directed to HSV. A description of HSV and certain exemplary features is provided below.

[0153] I. Herpes simplex virus (HSV) Herpes simplex viruses (HSVs) belong to the alpha subfamily of the human herpesvirus family and include HSV-1 and HSV-2. The structure of HSV-1 and HSV-2 primarily comprises (from inside to outside) a DNA core, capsid, tegument, and envelope. Each of HSV-1 and HSV-2 has a double-stranded DNA genome of approximately 153 kb, encoding at least 80 genes. The DNA core is enclosed by an icosahedral capsid composed of 162 capsomeres, 150 hexons, and 12 pentons, each composed of six different viral proteins. The DNA is surrounded by at least 20 different viral tegument proteins, which have structural and regulatory roles. Some of these proteins are involved in capsid transport to the nucleus and other organelles, viral DNA entry into the nucleus, activation of early gene transcription, inhibition of cellular protein biosynthesis, and mRNA degradation. The viral envelopes surrounding the tegument have at least 12 different glycoproteins (BN) on their surface. The glycoproteins can exist as heterodimers (H / L and E / I), with most existing as monomers.

[0154] HSV-1 and HSV-2 cause several mild, moderate, and severe conditions, including oral and genital ulcers, virus-induced blindness, viral encephalitis, and disseminated infection in newborns. HSV-1 and HSV-2 typically infect through different routes and affect different areas of the body, but the signs and symptoms they cause can overlap. Infections caused by HSV-1 represent one of the more widespread infections of the orofacial region and commonly cause herpes labialis, herpetic stomatitis, and keratitis. HSV-2 typically causes genital herpes and is primarily transmitted by direct sexual contact with lesions. While most genital HSV infections are caused by HSV-2, an increasing number of genital HSV infections are attributed to HSV-1. Genital HSV-1 infections are typically less severe and less common than genital HSV-2 infections.

[0155] HSV infection is transmitted through contact with herpes lesions, mucosal surfaces, genital secretions, or oral secretions. The average incubation period after exposure is typically 4 days but can range from 2 to 12 days. HSV particles can infect nerve extensions that service peripheral tissues and establish latency in these cells, namely the trigeminal ganglia and dorsal root ganglia in the sacral region, which can sporadically reactivate. Furthermore, like other herpesviruses, HSV infection is lifelong and generally asymptomatic. While not wishing to be bound by any particular theory, it is understood that HSV particles can be cleared from infected individuals independently of the development of clinical symptoms.

[0156] HSV infections are rarely fatal and are characterized by painful blisters that burst. There are few clear differences in clinical presentation based on the type of infecting virus. However, as noted above, HSV-1 infections tend to be less severe than HSV-2 infections, and patients infected with HSV-2 generally have more outbreaks.

[0157] A. Life Cycle As described herein, to initiate infection, HSV (HSV-1 or HSV-2) particles bind to the cell surface using viral glycoproteins and fuse their envelope with the cell membrane (see, e.g., Figure 2, step 1). After membrane fusion, the viral capsid and tegument proteins are internalized into the cytoplasm (see, e.g., Figure 2, step 2). Once inside the cytoplasm, the viral capsid accumulates in the nucleus and releases viral DNA into the nucleus (see, e.g., Figure 2, step 3). HSV replicates by three rounds of transcription that yield primarily α (immediate early) proteins that regulate viral replication, β (early) proteins that synthesize and package DNA, and γ (late) proteins, most of which are virion proteins (see Whitley et al., Lancet 2001 May 12;357(9267); Taylor et al., Front Biosci. 2002 Mar 1;7:d752-64; and Ibanez et al., Front Microbiol. 2018 Oct 11;9:2406, each of which is incorporated herein by reference in its entirety) (see, e.g., Figure 2, steps 4-6).

[0158] HSV capsids are assembled in the nucleus of infected cells (see, e.g., Figure 2, step 7). Once viral capsid assembly is complete in the nucleus, these particles continue their maturation process in this same compartment through the acquisition of tegument proteins. Upon leaving the nucleus, additional tegument proteins are added to the capsid. Meanwhile, glycoproteins are translated in the endoplasmic reticulum, glycosylated, processed in the trans-Golgi network (TGN), and then targeted to multivesicular bodies (see, e.g., Figure 2, step 8). They are then exported to the plasma membrane within early endosomes (see, e.g., Figure 2, step 9). The viral capsid in the cytoplasm then fuses with HSV-glycoprotein-containing endosomes to form infectious virions within vesicles (see, e.g., Figure 2, steps 10-12).

[0159] HSV (HSV-1 or HSV-2) can establish a latent infection. After primary infection, HSV either productively replicates within epithelial cells or enters sensory neuron axons and migrates to the neuronal nucleus. There, viral DNA remains as circular extrachromosomal DNA and does not have any lytic gene expression, but latency-associated transcripts are expressed and then spliced ​​to produce mRNA. This general transcriptional silence may allow the virus to remain hidden within the cell by evading immune surveillance. In some aspects, provided herein are technologies (e.g., compositions and methods) for enhancing, inducing, promoting, strengthening, and / or improving the immune response to HSV (e.g., HSV-1 and / or HSV-2) or components thereof (e.g., proteins or fragments thereof). In some embodiments, the technologies provided herein are designed to enhance, induce, promote, strengthen, and / or improve immune memory against HSV or components thereof (e.g., proteins or fragments thereof). In some embodiments, the technology described herein is designed to act as an immune boost to a primary vaccine, such as a vaccine directed against antigens and / or epitopes of HSV (e.g., HSV-1 and / or HSV-2).

[0160] The virus remains in this state for the life of the host or until an appropriate signal reactivates the virus and new progeny are generated, which then migrate through the neuronal axis to the site of primary infection and resume the lytic replication cycle.

[0161] B. HSV genome The genomes of both HSV-1 and HSV-2 are approximately 150 kb long double-stranded DNA that vary slightly between subtypes and strains. The genomes encode over 80 genes and have a high GC content: 67% and 69% for HSV-1 and HSV-2, respectively (see Whitley et al., Lancet 2001 May 12;357(9267); Taylor et al., Front Biosci. 2002 Mar 1;7:d752-64, and Jiao et al., Microbiol Resour Announc. 2019 Sep;8(39):e00993-19, which are incorporated herein by reference in their entireties).

[0162] The genome is organized as a unique long region (UL) and a unique short region (US). The UL is typically bounded by terminal long (TRL) and internal long (IRL) repeats. The US is typically bounded by terminal short (IRS) and internal short (TRS) repeats. Genes found in the unique region are present in the genome as a single copy, while genes encoded in the repeat region are present in two copies (see Whitley et al., Lancet 2001 May 12;357(9267); Taylor et al., Front Biosci. 2002 Mar 1;7:d752-64, and Jiao et al., Microbiol Resour Announc. 2019 Sep;8(39):e00993-19, which are incorporated herein by reference in their entireties).

[0163] HSV contains three origins of replication within its genome, named according to their location in either the long (oriL) or short (oriS) region of the genome. oriL is found as a single copy within the UL segment, while oriS is located within the repeat region of the short segment. Thus, oriL exists in two copies within the genome. Both oriL and oriS are palindromic sequences consisting of an AT-rich central region flanked by inverted repeats containing multiple binding sites of varying affinity for the virus-derived binding polypeptide (UL9). Either the oriL or one of the oriS sequences is sufficient for viral replication (see Whitley et.al., Lancet 2001 May 12;357(9267); Taylor et.al., Front Biosci. 2002 Mar 1;7:d752-64, and Jiao et.al., Microbiol Resour Announc. 2019 Sep;8(39):e00993-19, which are incorporated by reference in their entireties).

[0164] The viral genome also contains signals that regulate the proper processing of newly synthesized genomes for packaging into preformed capsids. Progeny genomes are generated in long concatemers that require cleavage into unit-length monomers. To this end, the viral genome contains two DNA sequence elements, pac1 and pac2, that ensure proper cleavage and packaging of unit-length progeny genomes. These elements are located within direct repeats (DRs) found within the inverted repeat regions at the ends of the viral genome (see Whitley et al., Lancet 2001 May 12;357(9267); Taylor et al., Front Biosci. 2002 Mar 1;7:d752-64; and Jiao et al., Microbiol Resour Announc. 2019 Sep;8(39):e00993-19, which are incorporated herein by reference in their entireties).

[0165] C. Specific HSV proteins ICP0 Herpes simplex virus 1 (HSV-1) infected cell protein 0 (ICP0) is an alpha (immediate early) protein of herpes simplex virus 1 that can activate HSV-1 gene expression, disrupt nuclear domain (ND)10 structure, mediate cellular protein degradation, and evade the host cell's inherent innate antiviral defenses (see Smith et al., Future Virol. 2011 Apr;6(4):421-429).

[0166] ICP22 Infected cell protein 22 (ICP22) is expressed from immediate-early (IE) genes during the replication cycle of HSV-1 and HSV-2. ICP22 can generally regulate viral and host gene transcription by altering the phosphorylation state of host RNA polymerase II (RNA pol II), and can also promote nuclear export complexes (NECs) that precisely localize to the nuclear membrane, facilitating nuclear bud formation (see Wu et al., Front Microbiol. 2021 Jun 7;12:668461).

[0167] VP16 The UL48 gene encodes VP16, or alpha gene transactivator (α-TIF). VP16 can activate transcription of viral immediate-early genes and is an important transactivator in the late stages of viral replication. Furthermore, VP16, as a tegument, is involved in virus assembly (see Fan, et al., Front Microbiol. 2020;11:1910).

[0168] During the early stages of viral infection, VP16 released by invading virions binds to immediate-early (IE) gene promoters and stimulates their transcription as a transactivator specifically acting on IE genes (see Fan, et al., Front Microbiol. 2020;11:1910). During the late stages, VP16 is assembled into the tegument to participate in virion assembly and promote their maturation (see Fan, et al., Front Microbiol. 2020;11:1910).

[0169] glycoproteins To replicate, the enveloped HSV must be able to fuse with the membrane of a living cell and deliver its genetic material into the cytoplasm. The HSV viral envelope surrounding the tegument contains at least 12 distinct glycoproteins (gB-gN) on its surface. The glycoproteins can exist as heterodimers (gH / gL and gE / gI), with most existing as monomers. HSV gC, gB, gD, gH, and gL are involved in the process of viral cell entry. Initial attachment is mediated by gC, followed by gD. Then, gH / gL pull the viral and cellular membranes together, and gB triggers membrane fusion. (Reske et al., Rev Med Virol. May-Jun 2007, and Arii et al., Adv Exp Med Biol. 2018;1045:3-21).

[0170] The present disclosure provides that, as further disclosed herein, HSV glycoprotein (e.g., gB, gC, gD, gE, gG, gH, gI, and / or gL) antigens and antigenic fragments thereof may be useful for the prevention or treatment of HSV, for example, in HSV antigen constructs and / or HSV compositions (e.g., immunogenic compositions, e.g., vaccines).

[0171] Glycoprotein C (gC) Mature HSV glycoprotein C (gC) is a 56 kDa protein that plays a role in initial cell attachment. Glycoprotein C is a type I membrane glycoprotein and is considered an important attachment protein and the main viral ligand for binding to heparin sulfate proteoglycans (HSPGs) on the cell surface. This binding can occur through gC interaction with HSPG-rich regions found on F-actin-rich membrane protrusions called filopodia.

[0172] Glycoprotein C has also been shown to be involved in regulating cell entry and infection by increasing the pH threshold for the acid-induced conformational change of gB. Low pH induces a reversible conformational change in gB domains I and V, functional regions containing hydrophobic loops important for cell fusion. By positively regulating the low-pH-induced conformational change of gB, gC can enhance the ability of HSV to enter cell types, such as epithelial cells, that require a low-pH mechanism for entry.

[0173] In addition to its role in adhesion, glycoprotein C has also been shown to play a role in immune evasion. Glycoprotein C is a target for lymphocyte cytotoxicity in certain cell types and can bind to complement component C3b, inhibiting complement activation. Furthermore, neutralizing epitopes present on other HSV glycoproteins, such as gB, can be protected by gC, preventing the immune response from blocking fusion.

[0174] Glycoprotein D (gD) HSV glycoprotein D (gD) is a 46-kDa type I membrane glycoprotein. The N-terminal ectodomain consists of 316 amino acids. Glycoprotein D promotes entry by interacting with several cell surface receptors, including herpesvirus entry mediator (HVEM), nectin-1 or nectin-2, and heparin sulfate containing specific modifications. These cell receptors do not function as coreceptors, as each glycoprotein interaction with the cellular receptor occurs independently of the other. Binding of gD to one of these cell receptors triggers a conformational change that converts gD from its autoinhibited closed state to an active state that transmits one of two signals thought to be required for gH / gL complex activation. HVEM, the first gD receptor identified, belongs to the tumor necrosis factor receptor family and is commonly found on T cells, B cells, dendritic cells, natural killer cells, macrophages, as well as non-immune cell types such as neurons and epithelial cells. Within the N-terminus of gD is a 37-residue hairpin structure that forms the entire site for binding to HVEM. Specifically, residues 1–32 of the N-terminal domain of gD bind to cysteine-rich domain 1 of HVEM. When not in contact with HVEM, this N-terminal extension adopts an extended, flexible conformation.

[0175] Clinical strains of HSV use nectin-1 for cell entry, whereas some variants of HSV utilize nectin-2. Furthermore, heparin sulfate is utilized by HSV-1 but not by HSV-2. Glycoprotein D interaction with nectin-1 has been shown to be essential in some cell types, such as neurons, even when other receptors are present on the cell surface.

[0176] Glycoprotein H (gH) / glycoprotein L (gL) complex Glycoprotein H (gH) is an essential 56 kD protein that exists as a heterodimeric complex with the 25 kDa glycoprotein L (gL) (a complex referred to herein as gH / gL). The gH / gL complex is required for cell fusion and entry. gH / gL does not share any structural similarity with documented fusion proteins and likely does not function as a cofusogen with gB. Instead, gH / gL may function as a regulator of fusion and a key component stabilizing the contact between HSV and the cell. Glycoprotein H receives a signal from gD via its H1 domain and transmits this signal to the membrane-proximal H3 domain, which in turn propagates the signal to the cytoplasmic tail of gH. When the cytoplasmic tail of gH receives this signal, it relieves the strain on the prefusion conformation of gB that favors attachment of the gB fusion loop to the cell surface, facilitating gB-mediated membrane fusion. Mutations in the C-terminal tail of gH have been shown to reduce fusion activity. Furthermore, antibody responses directed against gH have been shown to inhibit the fusion process mediated by gB-gH-gL. In addition to this essential role, gH contains an arginylglycylaspartic acid (RGD) motif that can bind to integrin receptors found on cells. The interaction of gH with integrins is thought to trigger intracellular signals that promote capsid transport.

[0177] Glycoprotein B (gB) Glycoprotein B has an apparent molecular weight of approximately 95-100 kDa and is composed of an extended rod- or spike-shaped ectodomain, a hydrophobic membrane-proximal region (MPR), a transmembrane region (TMR), and a C-terminal domain (CTD). While the ectodomain has been well characterized as an active participant in fusion, the MPR, TMR, and CTD may play roles in regulating fusion. Glycoprotein B is a class III fusogen. The glycoprotein B ectodomain architecture shares conformational similarity with fusogens from viruses not belonging to the Herpesviridae family. While glycoprotein B is activated by interaction with gH / gL, HSV cannot fuse with target cells by activation of gB alone and requires gB interaction with specific receptors to complete fusion. A known receptor target for gB is cell surface heparin sulfate, an interaction that is not essential for HSV fusion but is known to promote viral attachment to cell surfaces. Glycoprotein B can also interact with the paired immunoglobulin-like type 2 receptor, which is most commonly found on monocytes, macrophages, and dendritic cells.

[0178] HSV gB exists in two forms: pre-fusion and post-fusion. Several changes in the pre-fusion form of gB are thought to lead to its activity and post-fusion state. The first change occurs in domain V or MPR, which allows the fusion loop to move away from the viral membrane toward the cytoplasmic membrane. This change can generate a compressed intermediate conformation 1 that is not yet attached to the cytoplasmic membrane surface. The next change occurs in domain III, which involves gB adopting an extended intermediate conformation 2 that allows its fusion loop to attach to the cytoplasmic membrane surface. Finally, changes in domain V convert gB to a post-fusion conformation that favors membrane fusion.

[0179] The post-fusion form of HSV-1 gB has an ectodomain that exists as three promoters that interact to generate a rod-like trimeric structure. Each promoter is composed of five distinct domains with linker regions that individually form a hairpin shape. Each domain of an individual promoter interacts with the same domain of an adjacent promoter to form the described trimeric structure. Domain I houses the critical fusion loop and is commonly referred to as the fusion domain. Domain II facilitates interaction with gH / gL and is referred to as the gH / gL domain. Domain III is composed of alpha helices that help form the trimeric coil-coil central core of the protein. Domain IV, referred to as the crown domain, is located at the top of the post-fusion form and is thought to bind to cellular receptors. Antibodies that bind to the crown domain can inhibit gB binding to cellular receptors. Domain V consists of a long extension that connects the promoters together.

[0180] Glycoprotein E and glycoprotein I (gE / gI) Glycoprotein E is approximately 53 kDa, and glycoprotein I is approximately 141 kDa. Both proteins interact to form a heterodimeric complex (herein referred to as gE / gI) that plays a role in cell-to-cell spread and virus-induced fusion. Unlike gB, gD, and gH / gL, the gE / gI complex is not required for fusion and cell entry but is important for cell-to-cell spread. Inhibition of gE / gI formation affects HSV growth because this virus relies on cell-to-cell spread for its lytic cycle. The mechanism by which gE / gI promotes cell-to-cell spread is thought to depend on several tegument polypeptides. Cooperation between the tegument polypeptides UL11, UL16, and UL21 may play a role in gE processing, transport, and biological activity.

[0181] Glycoprotein G Glycoprotein G (gG) from both HSV-1 (gG1) and HSV-1 (gG2) was the first viral chemokine-binding protein shown to enhance cellular chemokine function. Glycoprotein G differs significantly in size between HSV-1 and HSV-2, measuring 76 kDa and 43 kDa, respectively. Glycoprotein G is unique in that its soluble form (gG2) can possess immunomodulatory capabilities through its extracellular activity. Once extracellular, gG2 binds to chemokines via their glycosaminoglycan (GAG)-binding domains without interfering with the chemokine's G protein-coupled receptor (GPCR) binding site. The interaction of gG2 with GAG-containing proteins can initiate the formation and accumulation of lipid rafts, which results in the clustering of chemokine receptors in these microdomains. The clustering of chemokine receptors then increases the local concentration of chemokines on the extracellular surface of host cells, allowing these chemokines to interact with GPCRs. This interaction likely leads to increased immune signaling responses and chemokine stimulation. This combination of receptor relocalization and presentation of chemokine complexes with SgG2 provides the molecular basis for enhanced chemokine function during HSV infection. This immune modulation contrasts with that seen in other viruses, where chemokine function is inhibited, in this case by SgG2. Without being bound by any particular theory, it is believed that global manipulation of endogenous immune signaling may be overall advantageous to HSV.

[0182] ICP47 Infected cell protein 47 (ICP47), encoded by the US12 gene, is a polymorphic protein that can block RNA splicing during early infection and then shuttle viral mRNA from the nucleus to the cytoplasm during late infection. ICP47 directly binds to the antigen-dependent transporter (TAP), restricting antigen transport and resulting in the generation of empty MHC-I (Cheng et al., Virol J. 2020 Jul 10;17(1):101). Binding of ICP47 to TAP stabilizes its inward conformation, thus blocking the translocation pathway and pointing to the endoplasmic reticulum (ER) cavity. By blocking viral antigen entry into the ER, HSV can evade the attack of cytotoxic T lymphocytes, leading to immune evasion and the establishment of a lifelong infection in host cells (Cheng et al., Virol J. 2020 Jul 10;17(1):101).

[0183] VHS The virion host shutoff (VHS) protein is a viral protein synthesized late in the course of viral infection and packaged into mature virion particles. Functionally, VHS is a viral RNase that preferentially degrades both host and viral mRNA species. VHS has been reported to interfere with dendritic cell (DC) activation in both productive and non-productive HSV infections (Cotter et al., J Virol. 2011 Dec;85(23):12662-12672).

[0184] US3 All members of the Alphaherpesviridae subfamily encode a serine / threonine kinase designated US3, a key virulence factor of herpes simplex virus type 1 (HSV-1) that plays multiple roles in the viral life cycle by phosphorylating multiple viral and cellular substrates (Kato et al., Adv Exp Med Biol. 2018;1045:45-62).

[0185] D. HSV vaccine Several HSV vaccines have been developed and evaluated in human clinical trials, primarily targeting HSV-2 and focusing primarily on the generation of neutralizing antibodies (nAbs) that target viral envelope glycoprotein D as a correlate of immune protection. See Table 1 below. Despite these vaccines demonstrating protection against HSV in preclinical and in some cases phase 2 studies, none of these vaccines have demonstrated sufficient efficacy for further development or commercialization.

[0186] The present disclosure provides insight that many previous strategies for developing pharmaceutical compositions (e.g., immunogenic compositions, e.g., vaccines) for treating HSV infection and / or protecting against viral infection have focused primarily, or even almost exclusively, on developing neutralizing antibodies that target surface glycoproteins. The present disclosure identifies problems with such strategies, including, for example, their failure to appreciate the value, or even importance, of ensuring that the induced immune response includes significant T cell activity (in some embodiments, CD4 T cell activity, in some embodiments, CD8 T cell activity, or in some embodiments, both). In some embodiments, for example, pharmaceutical compositions (e.g., immunogenic compositions, e.g., vaccines) that include or deliver CD4 and CD8 epitope(s) of one or more HSV antigens (e.g., HSV-1 antigens, HSV-2 antigens, or a combination thereof) in addition to one or more B cell antigens and / or epitopes may be used to treat and / or protect against HSV infection. [Table 1-1] [Table 1-2]

[0187] E. Antiviral treatment for HSV The present disclosure provides the recognition that the constructs and / or compositions described herein can be administered as part of a regimen with other therapeutic agents. The present disclosure also recognizes that a subject receiving the constructs and / or compositions described herein may have previously received other therapeutic agents.

[0188] In some embodiments, for example, the subject may be receiving or have previously received an antiviral agent against HSV. In some embodiments, the antiviral agent may be administered to treat HSV-1 or HSV-2 infection or recurrent episodes. In some embodiments, the antiviral agent is or includes acyclovir, valacyclovir, famciclovir, or a combination thereof. Table 2 below provides certain information regarding selected antiviral agents. [Table 2]

[0189] II. Constructs A.HSV antigen The present disclosure provides that HSV (e.g., HSV-1, HSV-2, or both) antigens and antigenic fragments thereof may be useful for the prevention or treatment of HSV, for example, in HSV antigen constructs and / or HSV compositions (e.g., immunogenic compositions, e.g., vaccines) further disclosed herein.

[0190] In some embodiments, HSV antigen fragments may be useful in HSV T cell antigen constructs. In some embodiments, HSV antigens (e.g., full-length HSV antigens) may be useful in HSV glycoprotein constructs.

[0191] In some embodiments, the polyribonucleotide encodes one or more HSV antigens or antigenic fragments thereof. In some embodiments, the polyribonucleotide encodes an HSV glycoprotein construct.

[0192] Numerous HSV antigens are known. The present disclosure provides polyribonucleotides encoding the HSV antigens described herein or antigenic fragments thereof. A summary of exemplary amino acid sequences of specific HSV (HSV-1, HSV-2, or both) proteins is provided in Tables 3-5 below. Exemplary amino acid sequences of specific HSV (HSV-1, HSV-2, or both) proteins is provided in Table 6 below. [Table 3] [Table 4] [Table 5] [Table 6-1] [Table 6-2] [Table 6-3] [Table 6-4] [Table 6-5] [Table 6-6] [Table 6-7] [Table 6-8] [Table 6-9] [Table 6-10] [Table 6-11]

Table 6-12

Table 6-13

Table 6-14

Table 6-15

Table 6-16

Table 6-17

Table 6-18

Table 6-19

Table 6-20

Table 6-21

Table 6-22

Table 6-23

Table 6-24

Table 6-25

Table 6-26

Table 6-27

Table 6-28

Table 6-29

Table 6-30

Table 6-31

Table 6-32

Table 6-33

Table 6-34

Table 6-35

Table 6-36

Table 6-37

Table 6-38

Table 6-39

Table 6-40

Table 6-41

Table 6-42

Table 6-43

Table 6-44

Table 6-45

[0193] In some embodiments, the present disclosure provides certain HSV antigen constructs (eg, HSV-1 antigen constructs, HSV-2 antigen constructs, or combinations thereof) that are particularly useful for effective vaccination.

[0194] In various embodiments, the HSV antigen construct comprises and / or encodes multiple HSV antigens (e.g., multiple HSV antigens that are or include one or more T cell and / or B cell antigens for HSV). As disclosed herein, T cell antigens include, for example, CD4 T cell antigens and / or CD8 T cell antigens. In some embodiments, the HSV antigen is a T cell antigen. In some embodiments, the HSV antigen is a B cell antigen.

[0195] In certain embodiments, the HSV antigen construct can comprise and / or encode at least one of UL1, UL21, UL27, UL29, UL39, UL40, UL46, UL47, UL48, UL49, RS1, RL2, UL5, UL9, UL19, UL25, UL30, UL52, US1, US7, US8, UL22, and / or UL54, or fragments thereof. In certain embodiments, the HSV antigen construct can comprise and / or encode at least one of UL1, UL21, UL27, UL29, UL39, UL40, UL46, UL47, UL48, and / or UL49, or fragments thereof. In certain embodiments, the HSV antigen construct can comprise and / or encode at least one of RS1, RL2, UL5, UL9, UL19, UL25, UL30, UL52, US1, US7, US8, UL22, and / or UL54 or fragments thereof.

[0196] In certain embodiments, the HSV antigen construct can comprise and / or encode a plurality (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19) of UL1, UL21, UL27, UL29, UL39, UL40, UL46, UL47, UL48, UL49, RS1, RL2, UL5, UL9, UL19, UL25, UL30, UL52, US1, US7, US8, UL22, and / or UL54, or fragments thereof. In certain embodiments, an HSV antigen construct can comprise and / or encode a plurality (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) of UL1, UL21, UL27, UL29, UL39, UL40, UL46, UL47, UL48, and / or UL49, or fragments thereof. In certain embodiments, an HSV antigen construct can comprise and / or encode a plurality (e.g., 1, 2, 3, 4, 5, 6, 7, 8, or 9) of RS1, RL2, UL5, UL9, UL19, UL25, UL30, UL52, US1, US7, US8, UL22, and / or UL54, or fragments thereof.

[0197] In some embodiments, the HSV antigen (e.g., a T cell or B cell antigen for HSV) is or comprises a UL1 polypeptide or a fragment thereof. In various embodiments, the UL1 polypeptide or fragment thereof has at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity) to a UL1 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of UL1 polypeptides known in the art include, but are not limited to, UL1 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the UL1 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NOs: 1, 2 and / or 3.

[0198] In some embodiments, the HSV antigen (e.g., a T cell or B cell antigen for HSV) is or comprises a UL21 polypeptide or fragment thereof. In various embodiments, the UL21 polypeptide or fragment thereof has at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity) to a UL21 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of UL21 polypeptides known in the art include, but are not limited to, UL21 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the UL21 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NOs: 4, 5 and / or 6.

[0199] The UL27 open reading frame encodes HSV gB (also referred to herein as the UL27 polypeptide). In some embodiments, the HSV antigen (e.g., a T cell or B cell antigen for HSV) is or includes a UL27 polypeptide or a fragment thereof. In various embodiments, the UL27 polypeptide or fragment thereof has at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity) to a UL27 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of UL27 polypeptides known in the art include, but are not limited to, UL27 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the UL27 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NOs: 7, 8, 9 and / or 74.

[0200] In some embodiments, the HSV antigen (e.g., a T cell or B cell antigen for HSV) is or comprises a UL29 polypeptide or fragment thereof. In various embodiments, the UL29 polypeptide or fragment thereof has at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity) to a UL29 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of UL29 polypeptides known in the art include, but are not limited to, UL29 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the UL29 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NOs: 10, 11 and / or 12.

[0201] In some embodiments, the HSV antigen (e.g., a T cell or B cell antigen for HSV) is or comprises a UL39 polypeptide or fragment thereof. In various embodiments, the UL39 polypeptide or fragment thereof has at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity) to a UL39 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of UL39 polypeptides known in the art include, but are not limited to, UL39 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the UL39 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NOs: 13, 14, and / or 15.

[0202] In some embodiments, the HSV antigen (e.g., a T cell or B cell antigen for HSV) is or comprises a UL40 polypeptide or fragment thereof. In various embodiments, the UL40 polypeptide or fragment thereof has at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity) to a UL40 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of UL40 polypeptides known in the art include, but are not limited to, UL40 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the UL40 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NOs: 16, 17 and / or 18.

[0203] In some embodiments, the HSV antigen (e.g., a T cell or B cell antigen for HSV) is or comprises a UL46 polypeptide or fragment thereof. In various embodiments, the UL46 polypeptide or fragment thereof has at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity) to a UL46 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of UL46 polypeptides known in the art include, but are not limited to, UL46 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the UL46 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NOs: 19, 20 and / or 21.

[0204] In some embodiments, the HSV antigen (e.g., a T cell or B cell antigen for HSV) is or comprises a UL47 polypeptide or fragment thereof. In various embodiments, the UL47 polypeptide or fragment thereof has at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity) to a UL47 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of UL47 polypeptides known in the art include, but are not limited to, UL47 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the UL47 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NOs: 22, 23 and / or 24.

[0205] In some embodiments, the HSV antigen (e.g., a T cell or B cell antigen for HSV) is or comprises a UL48 polypeptide or fragment thereof. In various embodiments, the UL48 polypeptide or fragment thereof has at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity) to a UL48 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of UL48 polypeptides known in the art include, but are not limited to, UL48 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the UL48 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NOs: 25, 26 and / or 27.

[0206] In some embodiments, the HSV antigen (e.g., a T cell or B cell antigen for HSV) is or comprises a UL49 polypeptide or fragment thereof. In various embodiments, the UL49 polypeptide or fragment thereof has at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity) to a UL49 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of UL49 polypeptides known in the art include, but are not limited to, UL49 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the UL49 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NOs: 28, 29, and / or 30.

[0207] In some embodiments, the HSV antigen (e.g., a T cell or B cell antigen for HSV) is or comprises an RS1 polypeptide or a fragment thereof. In various embodiments, the RS1 polypeptide or fragment thereof has at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity) to an RS1 amino acid sequence shown in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of RS1 polypeptides known in the art include, but are not limited to, RS1 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the RS1 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NOs: 31, 32, and / or 33.

[0208] In some embodiments, the HSV antigen (e.g., a T cell or B cell antigen for HSV) is or comprises an RL2 polypeptide or a fragment thereof. In various embodiments, the RL2 polypeptide or fragment thereof has at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity) to an RL2 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of RL2 polypeptides known in the art include, but are not limited to, RL2 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the RL2 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NOs: 34, 35, and / or 36.

[0209] In some embodiments, the HSV antigen (e.g., a T cell or B cell antigen for HSV) is or comprises a UL5 polypeptide or a fragment thereof. In various embodiments, the UL5 polypeptide or fragment thereof has at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity) to a UL5 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of UL5 polypeptides known in the art include, but are not limited to, UL5 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the UL5 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NOs: 37, 38 and / or 39.

[0210] In some embodiments, the HSV antigen (e.g., a T cell or B cell antigen for HSV) is or comprises a UL9 polypeptide or a fragment thereof. In various embodiments, the UL9 polypeptide or fragment thereof has at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity) to a UL9 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of UL9 polypeptides known in the art include, but are not limited to, UL9 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the UL9 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NO: 40, 41 and / or 42.

[0211] In some embodiments, the HSV antigen (e.g., a T cell or B cell antigen for HSV) is or comprises a UL19 polypeptide or fragment thereof. In various embodiments, the UL19 polypeptide or fragment thereof has at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity) to a UL19 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of UL19 polypeptides known in the art include, but are not limited to, UL19 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the UL19 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NO: 43, 44, and / or 45.

[0212] In some embodiments, the HSV antigen (e.g., a T cell or B cell antigen for HSV) is or comprises a UL25 polypeptide or a fragment thereof. In various embodiments, the UL25 polypeptide or fragment thereof has at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity) to a UL25 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of UL25 polypeptides known in the art include, but are not limited to, UL25 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the UL25 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NOs: 46, 47, and / or 48.

[0213] In some embodiments, the HSV antigen (e.g., a T cell or B cell antigen for HSV) is or comprises a UL30 polypeptide or fragment thereof. In various embodiments, the UL30 polypeptide or fragment thereof has at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity) to a UL30 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of UL30 polypeptides known in the art include, but are not limited to, UL30 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the UL30 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NOs: 49, 50, and / or 51.

[0214] In some embodiments, the HSV antigen (e.g., a T cell or B cell antigen for HSV) is or comprises a UL52 polypeptide or a fragment thereof. In various embodiments, the UL52 polypeptide or fragment thereof has at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity) to a UL52 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of UL52 polypeptides known in the art include, but are not limited to, UL52 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the UL52 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NO: 52, 53 and / or 54.

[0215] The US1 open reading frame encodes HSV gL (also referred to herein as the US1 polypeptide). In some embodiments, the HSV antigen (e.g., a T cell or B cell antigen for HSV) is or comprises a US1 polypeptide or a fragment thereof. In various embodiments, the US1 polypeptide or fragment thereof has at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity) to a US1 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of US1 polypeptides known in the art include, but are not limited to, US1 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the US1 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NOs: 58, 59, 60, and / or 61.

[0216] The US7 open reading frame encodes an HSV gI (also referred to herein as a US7 polypeptide). In some embodiments, the HSV antigen (e.g., a T cell or B cell antigen for HSV) is or comprises a US7 polypeptide or a fragment thereof. In various embodiments, the US7 polypeptide or fragment thereof has at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity) to a US7 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of US7 polypeptides known in the art include, but are not limited to, US7 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the US7 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NOs: 62, 63, 64, and / or 65.

[0217] The US8 open reading frame encodes HSV gE (also referred to herein as the US8 polypeptide). In some embodiments, the HSV antigen (e.g., a T cell or B cell antigen for HSV) is or comprises a US8 polypeptide or a fragment thereof. In various embodiments, the US8 polypeptide or fragment thereof has at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity) to a US8 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of US8 polypeptides known in the art include, but are not limited to, US8 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the US8 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NOs: 66, 67, 68, and / or 69.

[0218] The UL22 open reading frame encodes an HSV gH (also referred to herein as a UL22 polypeptide). In some embodiments, the HSV antigen (e.g., a T cell or B cell antigen for HSV) is or includes a UL22 polypeptide or a fragment thereof. In various embodiments, the UL22 polypeptide or fragment thereof has at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity) to a UL22 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of UL22 polypeptides known in the art include, but are not limited to, UL22 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the UL22 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NOs: 70, 71, 72 and / or 73.

[0219] In some embodiments, the HSV antigen (e.g., a T cell or B cell antigen for HSV) is or comprises a UL54 polypeptide or fragment thereof. In various embodiments, the UL54 polypeptide or fragment thereof has at least 80% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity) to a UL54 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of UL54 polypeptides known in the art include, but are not limited to, UL54 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the UL54 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NO: 55, 56 and / or 57.

[0220] In certain embodiments, the HSV antigen construct can comprise and / or encode one or more HSV antigens, including one or more T cell antigens against HSV of the present disclosure (e.g., CD4 and / or CD8 T cell antigens) and one or more HSV antigens that are not T cell antigens of the present disclosure. In certain embodiments, the HSV antigen construct can comprise and / or encode one or more HSV antigens, including one or more B cell antigens against HSV of the present disclosure and one or more HSV antigens that are not B cell antigens of the present disclosure. In certain embodiments, the HSV antigen construct can comprise and / or encode one or more HSV antigens, including one or more T cell antigens against HSV of the present disclosure and one or more HSV antigens that are B cell antigens against HSV (e.g., antigens that are or include B cell epitopes disclosed herein or otherwise known in the art). In certain embodiments, the HSV antigen construct can comprise and / or encode one or more HSV antigens, including one or more T cell antigens against HSV of the present disclosure and one or more HSV antigens selected from HSV glycoproteins or fragments thereof. In certain embodiments, the HSV antigen construct can comprise and / or encode one or more HSV antigens, including one or more T cell antigens for HSV of the present disclosure, and one or more HSV antigens selected from HSV gD protein or antigenic fragments thereof, HSV gB protein or antigenic fragments thereof, HSV gE protein or antigenic fragments thereof, HSV gG protein or antigenic fragments thereof, HSV gI protein or antigenic fragments thereof, HSV gH protein or antigenic fragments thereof, HSV gL protein or antigenic fragments thereof, HSV ICP4 protein or antigenic fragments thereof, or ICP8 protein or antigenic fragments thereof.

[0221] In various embodiments, the HSV antigen construct can be present in a composition for delivery of the HSV antigen construct to a subject. In various embodiments, the HSV antigen construct can be present in a composition for delivery of one or more HSV antigens and / or epitopes to a subject. In various embodiments, the HSV antigen construct is or includes an RNA molecule encoding one or more antigens and / or epitopes.

[0222] In certain embodiments, the HSV antigen construct and / or composition for delivering the HSV antigen construct may advantageously include, for example, one or more B cell antigens against HSV and one or more T cell antigens against HSV (e.g., CD4 and / or CD8 T cell antigens). Without wishing to be bound by any particular scientific theory or suggesting that other embodiments would also be advantageous, the combination of B cell antigens and T cell antigens may be advantageous in promoting immune system defense against HSV at multiple life cycle points, including, but not limited to, pre- and post-cellular entry.

[0223] Among other things, the present disclosure provides insight that many previous strategies for developing pharmaceutical compositions (e.g., immunogenic compositions, e.g., vaccines) for treating and / or protecting against viral infections have focused primarily, or even almost exclusively, on developing neutralizing antibodies that target surface glycoproteins. The present disclosure identifies problems with such strategies, including, for example, their failure to appreciate the value, or even importance, of ensuring that the induced immune response includes significant T cell activity (in some embodiments, CD4 T cell activity, in some embodiments, CD8 T cell activity, or in some embodiments, both).

[0224] Alternatively or additionally, the present disclosure provides insight that consideration of the expression of HSV proteins (e.g., during specific periods of the HSV life cycle and / or in specific tissues or compartments of an infected subject) can improve vaccine efficacy.

[0225] In some embodiments, the present disclosure provides techniques for identifying, selecting, and / or characterizing HSV protein sequences (e.g., HSV-1 protein sequences, HSV-2 protein sequences, or combinations thereof), and combinations thereof, that are particularly useful for inclusion in pharmaceutical compositions (e.g., immunogenic compositions, e.g., vaccines) described herein.

[0226] In some embodiments, pharmaceutical compositions (e.g., immunogenic compositions, e.g., vaccines) that include or deliver, for example, the CD4 and CD8 antigen(s) of one or more HSV proteins (e.g., HSV-1 proteins, HSV-2 proteins, or combinations thereof) in addition to one or more B cell antigens. Among other things, the present disclosure provides HSV antigen constructs (e.g., HSV-1 antigen constructs, HSV-2 antigen constructs, or combinations thereof) and compositions (e.g., pharmaceutical compositions, e.g., immunogenic compositions, e.g., vaccines) that include and / or deliver antigen constructs that induce both neutralizing antibodies and T cells (e.g., CD4 and / or CD8 T cells). Such neutralizing antibodies and T cells (e.g., CD4 and / or CD8 T cells) can target, for example, HSV glycoproteins, and in some embodiments, one or more additional HSV proteins. In some embodiments, the present disclosure provides such constructs and compositions that induce particularly strong neutralizing antibody responses and / or particularly diverse T cell responses (e.g., targeting multiple T cell antigens).

[0227] In some embodiments, the present disclosure provides such constructs and compositions that induce a robust B cell response, which in some embodiments includes the production of a diverse and specific repertoire of antibodies.

[0228] In some embodiments, the present disclosure provides such constructs and compositions that induce T cell and B cell responses against HSV antigens and / or epitopes.

[0229] The present disclosure provides the recognition that, for example, constructs and compositions comprising the RNA molecules described herein (e.g., encoding one or more HSV (e.g., HSV-1 and / or HSV-2) antigens and / or epitopes) can result in a higher degree of antigen presentation to various immune system components and / or pathways. In some embodiments, administration of such constructs or compositions can induce T cell and / or B cell responses. The present disclosure provides the insight that, for example, in some embodiments where T cell and B cell responses are induced in a subject, the subject may have a more sustained and long-lasting immune response. Such an immune response can be beneficial, for example, for preventing HSV (e.g., HSV-1 and / or HSV-2) reactivation with a single administration, which can increase vaccination rates and subject compliance compared to currently available vaccines that require administration every few years. In some embodiments, constructs and compositions comprising the RNA molecules described herein (e.g., encoding one or more HSV (e.g., HSV-1, HSV-2, or a combination thereof) antigens and / or epitopes) can provide more diverse protection (e.g., protection against HSV (e.g., HSV-1 and / or HSV-2) variants) because the constructs and compositions can induce multiple immune system responses, without wishing to be bound by any particular theory.

[0230] The present disclosure also provides the recognition that by administering constructs and compositions encoding HSV (e.g., HSV-1 and / or HSV-2) antigens and / or epitopes, the constructs and compositions described herein avoid administering HSV (e.g., HSV-1 and / or HSV-2) virions that can infect a subject, enter latency, and reactivate, causing a flare-up.

[0231] Furthermore, the present disclosure, in some embodiments, provides insights into how particularly effective pharmaceutical compositions (e.g., immunogenic compositions, e.g., vaccines) alter one or more characteristics of the innate immune system (and identify the source of problems with certain previous HSV vaccination strategies). The present disclosure provides certain such compositions, including, for example, compositions comprising RNA construct(s) encoding HSV (e.g., HSV-1 and / or HSV-2) protein(s) (e.g., HSV antigens or HSV epitopes), as described herein.

[0232] Separately, in some embodiments, the present disclosure provides particular pharmaceutical composition (e.g., immunogenic composition, e.g., vaccine) formats, including, for example, RNA pharmaceutical compositions (e.g., immunogenic compositions, e.g., vaccines) that include particular elements and / or sequences useful for vaccination.

[0233] The present disclosure provides various insights and techniques related to such HSV (eg, HSV-1 and / or HSV-2) antigen constructs and vaccine (eg, RNA vaccine) compositions.

[0234] As described herein, in many embodiments, provided compositions (e.g., pharmaceutical compositions, e.g., immunogenic compositions, e.g., vaccines) comprise an RNA active agent encoding one or more HSV (e.g., HSV-1 and / or HSV-2) polypeptides or antigenic fragments thereof; in some embodiments, such an RNA active agent is in a modified RNA format in that its uridine residues have been replaced with uridine analog(s), such as pseudouridine; alternatively or additionally, in some embodiments, such an RNA active agent comprises specific elements (e.g., cap, 5' UTR, 3' UTR, poly-A tail, etc.) and / or features (e.g., codon optimization) that have been identified, selected, characterized, and / or demonstrated to achieve significant (e.g., enhanced) translatability (e.g., in vitro) and / or expression (i.e., in a subject to which it is administered) of the encoded protein(s). Alternatively or additionally, in some embodiments, such RNA activity comprises specific elements and / or features that can be identified, selected, characterized, and / or demonstrated to achieve significant RNA stability and / or efficient production, particularly at large scales (e.g., 0.1-10 g, 10-500 g, 500 g-1 kg, 750 g-1.5 kg; one of skill in the art will understand that different products may be produced at different scales, e.g., depending on patient population size). In some embodiments, such RNA production scales can range from about 0.01 g / hr RNA to about 1 g / hr RNA, 1 g / hr RNA to about 100 g / hr RNA, about 1 g RNA / hr to about 20 g RNA / hr, or about 100 g RNA / hr to about 10,000 g RNA / hr. In some embodiments, such RNA production scales can range from tens or hundreds of milligrams to tens or hundreds of grams (or more) of RNA per batch.In some embodiments, such RNA manufacturing scale may allow for batch sizes in the range of about 0.01 g to about 500 g RNA, about 0.01 g to about 10 g RNA, about 1 g to about 10 g RNA, about 10 g to about 500 g RNA, about 10 g to about 300 g RNA, about 10 g to about 200 g RNA, or about 30 g to about 60 g RNA.

[0235] Still further, in many embodiments, provided compositions (e.g., pharmaceutical compositions, e.g., immunogenic compositions, e.g., vaccines) comprising an RNA active agent are prepared, formulated, and / or utilized in specific LNP compositions, as described herein.

[0236] Among other things, the present disclosure provides techniques for the rapid development of pharmaceutical compositions (e.g., immunogenic compositions, e.g., HSV vaccines) for delivering specific HSV (e.g., HSV-1 and / or HSV-2) antigenic constructs to a subject.

[0237] Additionally, the present disclosure provides, for example, nucleic acid constructs encoding the HSV (e.g., HSV-1 and / or HSV-2) antigens described herein, expressed HSV (e.g., HSV-1 and / or HSV-2) proteins, and various methods of production and / or use related thereto, as well as compositions developed therewith, and methods related thereto.

[0238] For example, the present disclosure provides techniques for preventing, characterizing, treating, and / or monitoring HSV (e.g., HSV-1 and / or HSV-2) outbreaks and / or infections, including various nucleic acid constructs and encoded proteins, as well as agents (e.g., antibodies) that bind to such proteins, and compositions containing and / or delivering them.

[0239] In some aspects, provided herein are technologies (e.g., compositions and methods) for enhancing, inducing, promoting, strengthening, and / or improving immune responses to HSV (e.g., HSV-1 and / or HSV-2) or components thereof (e.g., proteins or fragments thereof). In some embodiments, the technologies provided herein are designed to enhance, induce, promote, strengthen, and / or improve immunological memory to HSV (e.g., HSV-1 and / or HSV-2) or components thereof (e.g., proteins or fragments thereof). In some embodiments, the technologies described herein are designed to act as an immune boost to a primary vaccine, such as a vaccine directed against antigens and / or epitopes of HSV (e.g., HSV-1 and / or HSV-2). In some embodiments, compositions of the present disclosure comprise one or more polynucleotide constructs (e.g., one or more string constructs) encoding one or more antigens from HSV (e.g., HSV-1 and / or HSV-2). In some embodiments, the present disclosure provides vaccines or other compositions comprising nucleic acids encoding such HSV (e.g., HSV-1 and / or HSV-2) antigens. Those skilled in the art will understand from the context that when referring to a particular polynucleotide (e.g., DNA or RNA), "encoding" such antigen is actually referring to the coding strand or its complement.

[0240] The present disclosure provides pharmaceutical compositions (e.g., immunogenic compositions, e.g., vaccines) and related technologies (e.g., methods) for delivering specific HSV antigen constructs to a subject (e.g., a patient). In some embodiments, the present disclosure provides pharmaceutical compositions (e.g., immunogenic compositions, e.g., vaccines) and related technologies (e.g., methods) for delivering specific HSV-1 antigen constructs to a subject (e.g., a patient). In some embodiments, the present disclosure provides pharmaceutical compositions (e.g., immunogenic compositions, e.g., vaccines) and related technologies (e.g., methods) for delivering specific HSV-2 antigen constructs to a subject (e.g., a patient). In some embodiments, the present disclosure provides pharmaceutical compositions (e.g., immunogenic compositions, e.g., vaccines) and related technologies (e.g., methods) for delivering specific HSV-1 and HSV-2 antigen constructs to a subject (e.g., a patient).

[0241] The present disclosure further provides the recognition that some HSV antigens are common to both HSV-1 and HSV-2. The present disclosure also provides the recognition that some HSV antigens comprise sequences that are conserved between HSV-1 and HSV-2. Furthermore, the present disclosure recognizes that some HSV-1 antigens have, for example, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the equivalent HSV-2 antigen.

[0242] In some embodiments, the present disclosure provides certain HSV antigen constructs that are particularly useful for effective vaccination. In some embodiments, the HSV antigen construct is an HSV-1 antigen construct, an HSV-2 antigen construct, or a combination thereof.

[0243] Antigens utilized in accordance with the present disclosure are or include HSV (e.g., HSV-1 and / or HSV-2) components (e.g., antigenic fragments thereof, including epitopes that may include non-amino acid, e.g., carbohydrate moieties) that induce an immune response when administered to humans (or other animals, such as rodents and non-human primates susceptible to HSV (e.g., HSV-1 and / or HSV-2) infection).

[0244] In many embodiments, antigens utilized in provided pharmaceutical compositions (e.g., immunogenic compositions, e.g., vaccines) include both B cell and T cell antigens and / or epitopes described herein. In some specific embodiments, the delivered antigen includes both B cell and T cell (e.g., CD4 and / or CD8 T cell) antigens and / or epitopes, optionally included together within a single antigen polypeptide. In some embodiments, antigens utilized in provided pharmaceutical compositions (e.g., immunogenic compositions, e.g., vaccines) include T cell antigens and / or epitopes. In some embodiments, antigens utilized in provided pharmaceutical compositions (e.g., immunogenic compositions, e.g., vaccines) include B cell, CD4 T cell, and CD8 T cell epitopes together. Indeed, in some embodiments, the present disclosure defines and / or provides antigens comprising particularly useful epitopes for inclusion in HSV (e.g., HSV-1 and / or HSV-2) vaccines.

[0245] Exemplary antigens and / or epitopes for use in the compositions described herein include, for example, those provided in Tables 3-5 herein, and antigenic fragments thereof. In some embodiments, the exemplary antigens disclosed in Tables 3-5, and / or fragments and / or epitopes thereof, may be useful in the compositions described herein.

[0246] In some embodiments, provided pharmaceutical compositions (e.g., immunogenic compositions, e.g., HSV (e.g., HSV-1 and / or HSV-2) vaccines) comprise or deliver (e.g., cause expression in a recipient organism, e.g., by administration of a nucleic acid construct, such as an RNA construct described herein, encoding) an antigen that is or includes one or more epitopes (e.g., one or more B cell and / or one or more T cell antigens and / or epitopes) of an HSV (e.g., HSV-1 and / or HSV-2) protein. In some embodiments, the pharmaceutical compositions described herein induce an effective and relevant immune response against HSV (e.g., by targeting an HSV-1 protein, an HSV-2 protein, or a combination thereof).

[0247] In some embodiments, provided pharmaceutical compositions (e.g., immunogenic compositions, e.g., HSV (e.g., HSV-1 and / or HSV-2) vaccines) comprise or deliver an antigen that is or comprises a full-length HSV (e.g., HSV-1 and / or HSV-2) protein. In some embodiments, provided pharmaceutical compositions (e.g., immunogenic compositions, e.g., HSV (e.g., HSV-1 and / or HSV-2) vaccines) comprise or deliver an antigen that is or comprises a fragment of an HSV (e.g., HSV-1 and / or HSV-2) protein that is less than a full-length HSV (e.g., HSV-1 and / or HSV-2) protein. In some embodiments, provided pharmaceutical compositions (e.g., immunogenic compositions, e.g., HSV (e.g., HSV-1 and / or HSV-2) vaccines) comprise or deliver a chimeric polypeptide that is or comprises part or all of an HSV (e.g., HSV-1 and / or HSV-2) protein and one or more heterologous polypeptide elements.

[0248] In some embodiments, the antigen included in and / or delivered in a provided pharmaceutical composition (e.g., an immunogenic composition, e.g., an HSV (e.g., HSV-1 and / or HSV-2) vaccine) is or comprises one or more peptide fragments of an HSV (e.g., HSV-1 and / or HSV-2) antigen. In some such embodiments, each of the one or more peptide fragments comprises at least one epitope (e.g., one or more B-cell epitopes and / or one or more T-cell epitopes), which can be predicted, selected, evaluated, and / or characterized, for example, as described herein.

[0249] In some embodiments, antigens included in and / or delivered in provided pharmaceutical compositions (e.g., immunogenic compositions, e.g., HSV (e.g., HSV-1 and / or HSV-2) vaccines) are or include multiple peptide fragments of one or more HSV (e.g., HSV-1 and / or HSV-2) antigens. In some embodiments, a single polypeptide antigen can include multiple such fragments, presented, for example, as a series of antigens or fragments thereof described herein (e.g., a single polypeptide includes multiple amino acid sequences derived from different HSV antigens or fragments thereof, optionally separated by or otherwise associated with amino acid linkers or other intervening or terminal amino acid sequences). In some embodiments, a single RNA antigen construct can include multiple sequences encoding HSV antigens, presented, for example, as a series of antigen-coding sequences as described herein (e.g., a single RNA molecule includes multiple nucleic acid sequences encoding different HSV antigens or fragments thereof, optionally separated by or otherwise associated with nucleic acid linkers or other intervening or terminal nucleic acid sequences).

[0250] In some embodiments, one or more HSV (e.g., HSV-1 and / or HSV-2) antigens or antigenic fragments thereof may be linked to one or more sequences to which it is naturally linked. In some such embodiments, such sequence(s) may be or include one or more heterologous elements (e.g., one or more elements not naturally found in the relevant HSV (e.g., HSV-1 and / or HSV-2), e.g., a polypeptide or antigenic fragment thereof not naturally found directly bound to the relevant HSV (e.g., HSV-1 and / or HSV-2) antigen(s). For example, in some embodiments, antigenic peptides provided and / or utilized in accordance with the present disclosure may include one or more linker elements, one or more membrane-associated elements, one or more secreted elements, etc. In some embodiments, an antigenic polypeptide may include multiple HSV (e.g., HSV-1 and / or HSV-2) protein fragments or epitopes separated from one another by linkers.

[0251] In some embodiments, an HSV (e.g., HSV-1 and / or HSV-2) polypeptide, or fragment or epitope thereof, utilized in a construct described herein (or encoded by a polyribonucleotide described herein) may contain one or more sequence changes relative to a particular reference HSV (e.g., HSV-1 and / or HSV-2) polypeptide, or fragment or epitope thereof. For example, in some embodiments, the utilized antigen may contain one or more sequence mutations that are found or predicted to occur in circulating strains, e.g., in light of an assessment of sequence conservation and / or evolution of HSV (e.g., HSV-1 and / or HSV-2) polypeptides over time and / or strain. Alternatively or additionally, in some embodiments, the utilized antigen may contain one or more sequence mutations selected to, for example, affect the stability, folding, processing, and / or display of the antigen or any epitope thereof.

[0252] In some embodiments, an HSV (e.g., HSV-1 and / or HSV-2) polypeptide, or fragment or epitope thereof, utilized in an antigen described herein exhibits at least 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%, or 100% amino acid sequence identity with a relevant corresponding reference (e.g., wild-type) polypeptide, fragment, or epitope. In some embodiments, HSV (e.g., HSV-1 and / or HSV-2) polypeptides, or fragments or epitopes thereof, utilized in the antigens described herein exhibit at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence homology (i.e., identity or conservative substitution as understood in the art) amino acid sequence identity with the relevant corresponding reference (e.g., wild-type) protein, fragment, or epitope. Furthermore, in some embodiments, HSV (e.g., HSV-1 and / or HSV-2) polypeptides, or fragments or epitopes thereof, utilized in the antigens described herein, share conserved amino acid residues (e.g., at corresponding positions) with the relevant corresponding reference (e.g., wild-type) polypeptide, fragment, or epitope. Those skilled in the art will understand that, in general, a lower percent identity or homology can be tolerated for shorter peptides, since a single change, by definition, will have a greater impact on the percent identity or homology when considered relative to a smaller number of residues. For example, those skilled in the art will understand that for sequences of more than about 20 amino acids, the percent identity or homology will typically be greater than about 80%, and for sequences of more than about 50 amino acids, the percent identity or homology will typically be greater than about 90%.

[0253] In some embodiments, the degree of conservation may be assessed by considering the physicochemical differences between two amino acids, as described, for example, in WO2014 / 180569 (incorporated herein by reference in its entirety). It is well known in molecular evolution that frequently exchanged amino acids are likely to have chemical and physical similarities, whereas exchanged amino acids are rarely likely to have different physicochemical properties. The likelihood of a given substitution occurring naturally can be measured by a logarithmic matrix, relative to the likelihood of this substitution occurring by chance. The patterns observed in the logarithmic matrix imposed by natural selection "reflect the similarity of the functions of amino acid residues in weak interactions with each other in the three-dimensional conformation of a protein" (see Dayhoff et al., Atlas of protein sequence and structure 5:345, 1978, incorporated herein by reference in its entirety). In some embodiments, an evolutionary-based logarithmic matrix, which may be referred to as a "T-score," may be used to reflect the extent to which sequence variation may affect T cell recognition. Substitutions with a positive T score (i.e., log likelihood) are likely to occur naturally and therefore correspond to two amino acids with similar physicochemical properties. Substitutions with a positive T score are unlikely to alter immunogenicity. Conversely, substitutions with a negative T score reflect substitutions that are unlikely to occur naturally and therefore correspond to two amino acids with significantly different physicochemical properties. Such substitutions are more likely to alter immunogenicity. In some embodiments, the presence of a negative T score substitution in a sequence may indicate that it is relatively less useful in the vaccine antigens described herein, even if it is otherwise highly conserved.

[0254] In some embodiments, the utilized antigen induces an immune response that targets an HSV envelope glycoprotein. In some embodiments, one or more antigens induce an immune response that targets an HSV envelope glycoprotein. In some embodiments, the one or more antigens comprise one or more HSV protein sequences of antigens or epitopes of an HSV envelope glycoprotein (e.g., conserved sequences and / or sequences that are, or include, one or more B-cell epitopes and / or one or more CD4 epitopes and / or one or more CD8 epitopes). In some embodiments, the one or more antigens are, or include, an HSV gD protein, or a fragment or epitope thereof. In some embodiments, the one or more antigens are, or include, an HSV gB protein, or a fragment or epitope thereof. In some embodiments, the one or more antigens are, or include, an HSV gE protein, or a fragment or epitope thereof. In some embodiments, the one or more antigens are, or include, an HSV gG protein, or a fragment or epitope thereof. In some embodiments, one or more antigens are or comprise an HSV gI protein, or a fragment or epitope thereof. In some embodiments, one or more antigens are or comprise an HSV gE protein, or a fragment or epitope thereof. In some embodiments, one or more antigens are or comprise an HSV gH protein, or a fragment or epitope thereof. In some embodiments, one or more antigens are or comprise an HSV gL protein, or a fragment or epitope thereof. In some embodiments, one or more antigens are or comprise an HSV ICP4 protein, or a fragment or epitope thereof. In some embodiments, one or more antigens are or comprise an ICP8 polypeptide, fragment, or epitope thereof.

[0255] In various embodiments, the HSV antigen construct comprises and / or encodes multiple HSV antigens (e.g., multiple HSV antigens that are or include one or more T cell antigens for HSV) or fragments thereof provided in Table 3. In certain embodiments, the HSV antigen construct can comprise and / or encode at least one HSV antigen provided in Table 3, or a fragment thereof. In certain embodiments, the HSV antigen construct can comprise and / or encode at least one HSV antigen provided in Table 4, or a fragment thereof. In certain embodiments, the HSV antigen construct can comprise and / or encode at least one HSV antigen provided in Table 5, or a fragment thereof.

[0256] In certain embodiments, the HSV antigen construct can comprise and / or encode at least one T cell antigen against HSV, or a fragment thereof, provided in Table 3. In certain embodiments, the HSV antigen construct can comprise and / or encode at least one T cell antigen against HSV, or a fragment thereof, provided in Table 4. In certain embodiments, the HSV antigen construct can comprise and / or encode at least one T cell antigen against HSV, or a fragment thereof, provided in Table 5.

[0257] In certain embodiments, an HSV antigen construct can comprise and / or encode a plurality (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19) of HSV antigens, or fragments thereof, provided in Table 3. In certain embodiments, an HSV antigen construct can comprise and / or encode a plurality (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) of HSV antigens, or fragments thereof, provided in Table 4. In certain embodiments, an HSV antigen construct can comprise and / or encode a plurality (e.g., 1, 2, 3, 4, 5, 6, 7, 8, or 9) of HSV antigens, or fragments thereof, provided in Table 5.

[0258] In certain embodiments, an HSV antigen construct can comprise and / or encode a plurality (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19) T cell antigens against HSV selected from the HSV antigens provided in Table 3, or fragments thereof. In certain embodiments, an HSV antigen construct can comprise and / or encode a plurality (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) T cell antigens against HSV selected from the antigens provided in Table 4, or fragments thereof. In certain embodiments, an HSV antigen construct can comprise and / or encode a plurality (e.g., 1, 2, 3, 4, 5, 6, 7, 8, or 9) T cell antigens against HSV selected from the antigens provided in Table 5, or fragments thereof.

[0259] In some embodiments, the antigen utilized in accordance with the present disclosure is 1) HLA-I or HLA-II binding (e.g., to the HLA allele(s) present in the relevant population) 2) HLA ligandomics data, optionally confirmed by mass spectrometry 3) Relatively high expression 4) Sequence preservation 5) Surface exposure 6) Seroreactivity 7) Immunogenicity (e.g., the presence of one or more B cell and / or T cell antigens and / or epitopes, e.g., evidence of the ability to induce sterile protection in model systems including humans, non-human primates, and / or mice). 8) HSV (e.g., HSV-1 and / or HSV-2) protein sequences identified and / or characterized by one or more of the following: the absence of sequences overlapping with the human proteome.

[0260] In some embodiments, such properties are evaluated experimentally or computationally, hi some embodiments, such properties are evaluated by consultation with published reports.

[0261] For example, in some embodiments, HLA-I and / or HLA-II binding is assessed experimentally, and in some embodiments, predicted.

[0262] In some embodiments, predicted HLA-I or HLA-II binding is assessed using algorithms such as neonmhc1 and / or neonmhc2, which predict and / or characterize MHC class I and MHC class II binding potential, respectively. Alternatively or additionally, in some embodiments, the MHC-peptide presentation prediction algorithm or MHC-peptide presentation predictor is or includes NetMHCpan or NetMHCIIpan. In some embodiments, a hidden Markov model approach may be utilized for MHC-peptide presentation prediction and / or characterization. In some embodiments, the peptide prediction model MARIA may be utilized. In some embodiments, NetMHCpan is not utilized to predict or characterize MHC binding potential for peptides as described herein. In some embodiments, the peptide prediction model MARIA may be utilized. In some embodiments, NetMHCIIpan is not utilized to predict or characterize MHC binding potential for peptides as described herein. In some embodiments, neither NetMHCpan nor NetMHCIIpan is utilized to predict or characterize MHC binding potential for peptides as described herein. In some embodiments, the MHC-peptide presentation prediction algorithm or predictor is or includes RECON® (Real-time Epitope Computation for ONcology), which provides high-quality MHC-peptide presentation predictions based on expression, processing, and binding capacity. See, e.g., Abelin et al., Immunity 21:315, 2017; Abelin et al., Immunity 15:766, 2019.

[0263] In some embodiments, HLA binding and / or ligandomics assessment takes into account the geographic region of the subject to be immunized. For example, in some embodiments, the diversity of HLA alleles is taken into account. In some embodiments, the antigen(s) included in the provided pharmaceutical compositions (e.g., immunogenic compositions, e.g., vaccines) are or include peptides (e.g., epitopes) that, when considered together, are predicted or determined to bind to a significant percentage (e.g., at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or more) of the HLA alleles predicted or known to be present in the relevant region or population. In some embodiments, the antigen(s) included in the provided pharmaceutical compositions (e.g., immunogenic compositions, e.g., vaccines) are or include peptides that, when considered together, are predicted or determined to bind to the most common (e.g., the 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 most common, or at least 1, 2, 3, 4, or 5 of the 10 most common, etc.). HLA alleles expected or known to be present in the relevant region or population).

[0264] In some embodiments, the expression level is experimentally determined (e.g., in a model system or in infected humans). In some embodiments, the expression level is a reported level (e.g., in a published or presented report). In some embodiments, the expression level is assessed as RNA (e.g., via RNASeq). In some embodiments (and typically preferably), the expression level is assessed as protein.

[0265] In some embodiments, sequence conservation is assessed, for example, using publicly available sequence evaluation software (e.g., multiple sequence alignment programs MAFFT, Clustal Omega, etc.). In some embodiments, sequence conservation is determined by consultation with publicly available resources (e.g., sequences). In some embodiments, sequence conservation includes consideration of currently or recently detected strains (e.g., in active outbreaks).

[0266] In some embodiments, surface exposure is assessed by reference to publicly available databases and / or software.

[0267] In some embodiments, seroreactivity is assessed by contacting a serum sample from an infected individual with a polypeptide comprising a sequence of interest (which may be displayed, for example, via phage display or peptide arrays; see, e.g., Whittemore et al., "A General Method to Discover Epitopes from Sera," PlosOne, 2016; https: / / doi.org / 10.1371 / journal.pone.0157462). In some embodiments, seroreactivity is assessed by consultation with literature reports and / or database data indicating serorecognition sequences.

[0268] In some embodiments, assessment of the presence of immunoreactivity and / or epitopes may be or include consultation with the Immune Epitope Database (IEDB), which those skilled in the art will recognize as a freely available resource funded by NIAID that catalogs experimental data on antibody and T-cell epitopes (see iedb.org).

[0269] In some embodiments, the antigen(s) utilized in accordance with the present disclosure are characterized by dendritic cell presentation, which may exhibit HLA binding and / or immunogenicity.

[0270] In some embodiments, the antigen(s) utilized in accordance with the present disclosure are or include sequences (e.g., epitopes, fragments, full proteins) of HSV proteins found in the HSV envelope. In some embodiments, the antigen(s) utilized in accordance with the present disclosure are or include sequences (e.g., epitopes, fragments, full proteins) of HSV proteins found in the HSV tegument.

[0271] Among other things, the present disclosure provides the insight that, in some embodiments, it may be desirable to optionally include two or more different epitopes from two or more different HSV (e.g., HSV-1 and / or HSV-2) proteins in a pharmaceutical composition (e.g., an immunogenic composition, e.g., a vaccine) that may be useful in treating HSV. [Table 7-1] [Table 7-2] [Table 7-3]

[0272] 1. Exemplary Antigen Formats In some embodiments, the antigen utilized as described herein is or comprises a full-length viral protein. In some embodiments, the antigen utilized as described herein is or comprises a fragment or domain of a viral polypeptide, or an antigenic fragment thereof. In some embodiments, the antigen utilized as described herein is a membrane-bound antigen (e.g., an antigenic fragment thereof fused to a membrane-bound portion, e.g., a transmembrane portion). In some embodiments, the provided pharmaceutical compositions (e.g., immunogenic compositions, e.g., vaccines) comprise or deliver antigenic sequences that are or comprise one or more antibody epitopes and / or one or more CD4 T cell and / or CD8 T cell epitopes.

[0273] In some embodiments, antigens utilized as described herein include one or more variant sequences relative to a related reference antigen. For example, in some embodiments, protease cleavage sites are removed or blocked, and alternatively or additionally, in some embodiments, truncated antigens are utilized.

[0274] In some embodiments, an antigen utilized as described herein comprises a multimerizing element (eg, a heterologous multimerizing element).

[0275] In some embodiments, an antigen utilized as described herein comprises a membrane-associated element (eg, a heterologous membrane-associated element), such as a transmembrane domain.

[0276] In some embodiments, an antigen utilized as described herein comprises a secretory signal (eg, a heterologous secretory signal).

[0277] In some embodiments, the sequence utilized may be longer (and therefore contain more epitopes) than the viral protein found in nature.

[0278] In some embodiments, the sequences utilized may be derived from different strains or multiple strains (e.g., may be circulating and / or otherwise associated within a population to which the pharmaceutical composition (e.g., immunogenic composition, e.g., vaccine) is administered).

[0279] In some embodiments, antigens utilized as described herein may include multiple epitopes (e.g., B cell and / or T cell antigens and / or epitopes) arranged in a non-natural arrangement (e.g., a string construct as described herein). In some embodiments, antigens utilized as described herein may include multiple epitopes predicted or demonstrated to bind to HLA alleles reflective of the population to which a pharmaceutical composition (e.g., an immunogenic composition, e.g., a vaccine) composition will be administered as described herein.

[0280] In some embodiments, provided pharmaceutical compositions (e.g., immunogenic compositions, e.g., vaccines) may comprise or deliver one or more antigens that comprise a B cell epitope and one or more antigens that comprise a T cell epitope. In some embodiments, provided pharmaceutical compositions (e.g., immunogenic compositions, e.g., vaccines) may comprise or deliver one antigen that comprises both a B cell and a CD4 epitope and a separate antigen that comprises a CD8 epitope.

[0281] As described herein, in some embodiments, the provided techniques involve administering multiple antigens to the same subject. In some embodiments, the multiple antigens are administered simultaneously (e.g., in a single dose). In some embodiments, different antigens may be administered at different times (e.g., different doses, e.g., a prime dose versus a boost dose). In some embodiments, the multiple antigens are administered via the same composition.

[0282] For clarity, a single "antigen" polypeptide may contain multiple "epitopes," which may or may not be inherently linked to one another. For example, a single string construct antigen may contain multiple epitopes that may be derived from different portions of the same HSV (e.g., HSV-1 and / or HSV-2) protein and / or different HSV (e.g., HSV-1 and / or HSV-2) proteins, linked together as described herein in a single polypeptide.

[0283] Thus, a single pharmaceutical composition (e.g., an immunogenic composition, e.g., a vaccine) described herein may contain or deliver a single antigen (e.g., because the pharmaceutical composition comprises a nucleic acid, such as RNA, that encodes the antigen and that is expressed upon administration), which itself may comprise multiple epitopes (either in their natural arrangement relative to each other, or in an engineered or constructed arrangement as described herein), or may contain or deliver multiple antigens, each of which may similarly be or comprise a single epitope or multiple epitopes (either in their natural arrangement relative to each other, or in an engineered or constructed arrangement as described herein). Furthermore, a single pharmaceutical composition (e.g., an immunogenic composition, e.g., a vaccine) may contain, for example, multiple different nucleic acids (e.g., RNAs) each encoding a different antigen(s), or in some embodiments, may comprise a single nucleic acid that encodes (and expresses) multiple antigens. Furthermore, a single pharmaceutical composition (e.g., an immunogenic composition, e.g., a vaccine) comprising multiple different nucleic acids (e.g., RNAs) encoding antigens can be prepared in some embodiments by mixing the RNAs and then incorporating the mixture into LNPs, or alternatively, by formulating individual RNAs into LNPs and then mixing the LNPs. In some embodiments, the mixture (preparation of LNP precursor RNAs, or mixture of LNPs) can contain related RNAs in a 1:1 ratio or other ratio that is preferred (e.g., to achieve a desired relative presentation of antigens or epitopes) in a subject to which the composition is administered.

[0284] In certain embodiments, a pharmaceutical composition (e.g., an immunogenic composition, e.g., a vaccine) comprises or can deliver a combination comprising a polypeptide or fragment thereof encoded by all or a portion of UL1, UL21, UL27, UL29, UL39, UL40, UL46, UL47, UL48, UL49, RS1, RL2, UL5, UL9, UL19, UL25, UL30, UL52, US1, US7, US8, UL22, and / or UL54 or a fragment thereof.

[0285] In some embodiments, provided compositions comprise or deliver an HSV (e.g., HSV-1 and / or HSV-2) envelope glycoprotein antigen (e.g., a full-length HSV (e.g., HSV-1 and / or HSV-2) envelope glycoprotein, a fragment thereof, or one or more epitopes thereof, e.g., a string construct). In some embodiments, provided compositions comprise or deliver such an HSV (e.g., HSV-1 and / or HSV-2) envelope glycoprotein antigen along with one or more B-cell targets (e.g., epitopes), which may be or may include, for example, one or more other HSV (e.g., HSV-1 and / or HSV-2) proteins (or fragments or epitopes thereof). In some embodiments, such B-cell targets are or include HSV (e.g., HSV-1 and / or HSV-2) proteins (or fragments or epitopes thereof) predicted or known to induce a B-cell response in infected humans. For example, in some embodiments, the B cell target is or comprises an HSV (e.g., HSV-1 and / or HSV-2) protein (or fragment or B cell epitope thereof) to which sera from infected individual(s) are reactive. In some particular embodiments, the B cell target is or comprises an HSV (e.g., HSV-1 and / or HSV-2) envelope glycoprotein or other related HSV (e.g., HSV-1 and / or HSV-2) protein, or fragment or epitope thereof.

[0286] In some embodiments, provided compositions comprise or deliver a string construct antigen comprising multiple T cell epitopes, optionally from two or more HSV (e.g., HSV-1 and / or HSV-2) proteins. In some such embodiments, provided compositions further comprise or deliver one or more B cell targets. Alternatively or additionally, in some embodiments, the string construct antigen so utilized comprises an HSV (e.g., HSV-1 and / or HSV-2) sequence (e.g., one or more fragments or epitopes, e.g., T cell epitopes and / or B cell epitopes, but in some embodiments specifically T cell epitopes).

[0287] In some embodiments, the string construct antigen comprises both B cell epitopes and T cell epitopes (optionally from the same HSV (e.g., HSV-1 and / or HSV-2) protein or from different HSV (e.g., HSV-1 and / or HSV-2) proteins).

[0288] In some embodiments, different antigens may be delivered by administration of different compositions, and thus, in some embodiments, they may be administered simultaneously (e.g., as an admixture or otherwise substantially simultaneously), and in some embodiments, at different times. As just one example, in some embodiments, a particular antigen or antigen(s) may be delivered via an initial pharmaceutical composition (e.g., immunogenic composition, e.g., vaccine) dose, and one or more other antigen(s) may be delivered via one or more booster dose(s).

[0289] 2. Exemplary Multiple Epitope Antigens In some embodiments, the antigen utilized in (i.e., contained in and / or otherwise delivered by) a pharmaceutical composition (e.g., an immunogenic composition, e.g., a vaccine) described herein comprises multiple epitopes, e.g., epitopes of a single HSV (e.g., HSV-1 and / or HSV-2) protein or multiple proteins.

[0290] In some embodiments, an antigen may comprise two or more epitopes from the same HSV (e.g., HSV-1 and / or HSV-2) protein, which may be in their natural configuration relative to each other (e.g., within a fragment, in the case of related proteins). However, in some embodiments, an antigen may comprise at least two epitopes configured in a non-natural relationship relative to each other (e.g., comprised in a string construct described herein).

[0291] Among other things, the present disclosure provides the insight that string construct antigens may be particularly useful or effective for vaccination against HSV (e.g., HSV-1 and / or HSV-2) infection. Without wishing to be bound by any particular theory, the present disclosure proposes that the ability to link individual epitopes predicted or determined to have particular attributes, such as binding to relevant HLA alleles, expression during relevant infection, and particularly conserved sequence representation, potentially across multiple different HSV (e.g., HSV-1 and / or HSV-2) proteins, may prove uniquely beneficial, or indeed important, for effective vaccination against HSV (e.g., HSV-1 and / or HSV-2), where more conventional vaccination approaches have thus far provided only limited protection.

[0292] In some embodiments, multi-epitope antigens (e.g., string construct antigens or polyepitope antigens) may be administered as polypeptides and / or as collections of peptides. Alternatively or additionally, multi-epitope antigens may be administered as preparations of cells containing (e.g., expressing) the antigens. However, the present disclosure further provides the insight that in some embodiments, delivery by administration of nucleic acids, particularly RNA encoding multi-epitope antigens, may be particularly useful and / or effective.

[0293] As noted elsewhere herein, experience with SARS-CoV-2 has demonstrated that RNA administration can be a particularly effective method for delivering infectious disease antigens. Furthermore, the present disclosure provides insight that various features of nucleic acid formats, including, for example, their flexibility and adaptability to rapid design and modification, including the incorporation of various insights (e.g., bioinformatics input, etc.), make them particularly attractive for use in HSV (e.g., HSV-1 and / or HSV-2) vaccines. In particular, the present disclosure provides insight that, in some embodiments, administration of RNA encoding the string construct antigens described herein can be a particularly desirable and / or effective approach to immunization against HSV (e.g., HSV-1 and / or HSV-2) infection.

[0294] In some embodiments, the "string" polynucleotide sequence encodes multiple antigens and / or epitopes in tandem. In some embodiments, the string encodes about 2 to about 100, about 2 to about 75, about 2 to about 50, about 2 to about 25, about 2 to about 20, about 2 to about 15, about 2 to about 10, or about 2 to about 5 antigens and / or epitopes. In some embodiments, the string encodes about 5 to about 100, about 5 to about 75, about 5 to about 50, about 5 to about 25, about 5 to about 20, about 5 to about 15, or about 5 to about 10 antigens and / or epitopes. In some embodiments, the "string" polynucleotide sequence encodes multiple epitopes in tandem. In some embodiments, the string encodes about 2 to about 1,000, or about 2 to about 10,000 antigens and / or epitopes. In some embodiments, approximately 2 to 5,000 antigens and / or epitopes are encoded by a single polynucleotide string. In some embodiments, approximately 2 to 4,000 antigens and / or epitopes are encoded by a single polynucleotide string. In some embodiments, approximately 2 to 3,000 antigens and / or epitopes are encoded by a single polynucleotide string. In some embodiments, approximately 2 to 2,000 antigens and / or epitopes are encoded by a single polynucleotide string. In some embodiments, approximately 2 to 1,000 antigens and / or epitopes are encoded by a single polynucleotide string. In some embodiments, approximately 10 to 500 antigens and / or epitopes are encoded by a single polynucleotide string. In some embodiments, approximately 10 to 200 antigens and / or epitopes are encoded by a single polynucleotide string. In some embodiments, approximately 20 to 100 antigens and / or epitopes are encoded by a single polynucleotide string.

[0295] In some embodiments, the epitopes encoded by the string construct include epitopes predicted by HLA binding and presentation prediction software to be likely to be presented to T cells by HLA-encoded proteins to elicit an immune response. In some embodiments, the epitopes predicted to be likely to be presented by HLA-encoded proteins are selected from any one of the proteins or peptides listed in Tables 3-5. In some embodiments, the epitopes in the string construct include epitopes that are membrane-associated or otherwise accessible, for example, at relevant time(s) during the HSV (e.g., HSV-1 and / or HSV-2) life cycle.

[0296] In some embodiments, antigens utilized in accordance with the present disclosure are or include UL1, UL21, UL27, UL29, UL39, UL40, UL46, UL47, UL48, UL49, RS1, RL2, UL5, UL9, UL19, UL25, UL30, UL52, US1, US7, US8, UL22, and / or UL54, or fragments thereof, variants thereof, and / or fragments or epitopes of any of the foregoing, and combinations of any of the foregoing. In some embodiments, antigens utilized in accordance with the present disclosure are or include HSV proteins, such as HSV envelope protein, HSV tegument protein, HSV membrane protein, and variants thereof, and / or fragments or epitopes of any of the foregoing, and combinations of any of the foregoing. In some embodiments, string constructs can include multiple epitopes from two, three, four, or more HSV proteins. In some embodiments, string constructs include one or more features described herein, including the Examples and Tables. In some embodiments, the string construct comprises or is encoded by a sequence shown in Tables 3-5.

[0297] Alternatively or additionally, in some embodiments, one or more string constructs may include one or more other epitopes (e.g., predicted or documented in the literature). In some embodiments, the string construct may include sequences encoding features such as a linker and a cleavage site (e.g., a self-cleavage site such as a T2A or P2A sequence). In some embodiments, linkers enriched with G and S residues may be used. In some embodiments, an exemplary linker may have the sequence GGGGSGGGGS (SEQ ID NO: 167) or GGSGGGGSGG (SEQ ID NO: 165).

[0298] In some embodiments, the string construct comprises two or more overlapping epitope sequences.

[0299] In some embodiments, the string construct comprises or is encoded by a sequence that is 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to any one of the sequences in Tables 3-5. As noted above, when the sequences being compared are longer than about 20 amino acids, the percent identity or homology is typically greater than about 80%, and when the sequences are longer than about 50 amino acids, the percent identity or homology is typically greater than about 90%.

[0300] In some embodiments, epitopes are positioned on the string to maximize the immunogenicity of the string, e.g., by maximizing recognition by the subject's HLA allele repertoire, hi some embodiments, the same string encodes epitopes that can and / or are predicted to bind to different HLA alleles. For example, as well exemplified in the Sequence Listing, and as shown in at least Tables 3-5, the string can encode epitope(s) including (a) a first epitope that binds or is predicted to bind to a first MHC peptide encoded by a first HLA allele, (b) a second epitope that binds or is predicted to bind to a second MHC peptide encoded by a second HLA allele, and (c) a third epitope that binds or is predicted to bind to a third MHC peptide encoded by a third HLA allele, and further such epitopes can be added, for example, as in the string sequences provided herein, wherein the first, second, and third epitopes are epitopes derived from the same HSV (e.g., HSV-1 and / or HSV-2) protein or different HSV (e.g., HSV-1 and / or HSV-2) proteins. In this way, the distribution of epitopes encoded by a single string is maximized for presentation to different MHC-based T cells, thereby maximizing the probability of generating a desired immune response from a wider range of patients in a given population and the robustness of the response in each patient.

[0301] In some embodiments, epitopes included in string constructs are selected based on high-scoring predictions for HLA binding by reliable prediction algorithms or systems, such as the RECON prediction algorithm. In some embodiments, the present disclosure offers the prospect that selecting epitopes based on highly reliable and efficient prediction algorithms in the placement of epitopes encoded by strings, with or without non-epitope sequences or sequences flanking the epitope, can provide particularly successful strings whose immunogenicity is validated in ex vivo cell culture models or animal models, specifically by demonstrating T cell induction after vaccination with the string construct or a polypeptide encoded by the string construct, with the observation of an epitope-specific T cell response. In some embodiments, validation can be derived from use in human patients, or by the observation that T cells obtained from patients after vaccination demonstrate an epitope-specific, efficient, and sustained T cell response. In some embodiments, the efficacy of a string as a vaccine is influenced by its design, which depends in part on, among other things, the strength of the bioinformatics information used in judicious execution of the design, the reliability of MHC presentation prediction models, and the efficiency of epitope processing when the string vaccine is expressed intracellularly.

[0302] In some embodiments, such multi-epitope RNA (e.g., mRNA) constructs comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or more antigens and / or epitopes. In some embodiments, the pharmaceutical composition comprises 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more strings. In some embodiments, the pharmaceutical composition comprises 6 strings. In some embodiments, the pharmaceutical composition comprises 7 strings. In some embodiments, the pharmaceutical composition comprises 8 strings. In some embodiments, the pharmaceutical composition comprises 9 strings. In some embodiments, the pharmaceutical composition comprises 10 strings.

[0303] In some embodiments, the epitope-encoding sequence in the string construct is flanked by one or more sequences selected for greater immunogenicity, better cleavage for peptide presentation to MHC, better expression, and / or improved translation in cells in a subject. In some embodiments, the flanking sequences include linkers with specific cleavable sequences. In some embodiments, the epitope-encoding sequence in the string construct is flanked by secreted protein sequences.

[0304] In some embodiments, the string sequence encodes an epitope that may include or be otherwise linked to a signal sequence, such as those listed in Table 7, or a sequence having at least 1, 2, 3, 4, or 5 amino acid differences therefrom. In some embodiments, the string sequence encodes an epitope that may include or be otherwise linked to a signal sequence, such as MFVFLVLLPLVSSQCVNLT (SEQ ID NO: 90), or at least a sequence having 1, 2, 3, 4, or at most 5 amino acid differences therefrom. In some embodiments, the string sequence encodes an epitope that may be linked at the N-terminus by a sequence enriched in G and S residues, or a sequence having 1, 2, 3, 4, or at most 5 amino acid differences therefrom. In some embodiments, an exemplary linker that may be useful for linking epitopes has the sequence GGSGGGGSGG (SEQ ID NO: 165).

[0305] In some embodiments, the linked sequences may include a linker with a cleavage sequence, for example, with a specific cleavable sequence.

[0306] In some embodiments, the string construct is linked to a transmembrane domain (TM) or other membrane-associated element. In some embodiments, the linker may have a length of 2, 3, 4, 5, 6, 7, 8, 9, 10, or more amino acids. In some embodiments, linkers of about 30 or less, 25, 20, 15, 10, or fewer amino acids are used. In some embodiments, the linker sequence is not limited to containing any particular amino acid; in some embodiments, the linker sequence comprises any amino acid. In some embodiments, the linker or cleavage sequence comprises glycine (G). In some embodiments, the linker or cleavage sequence comprises serine (S). In some embodiments, the linker is designed to contain amino acids based on cleavage predictors to generate peptide sequences with highly cleavable sequences, a novel and effective method for delivering immunogenic T cell epitopes in a T cell vaccine setting.

[0307] In some embodiments, the distribution of epitopes encoded in the string construct and their juxtaposition are designed to promote cleavage sequences contributed by the amino acid sequence of the epitope and / or flanking or linking residues, thereby using minimal linker sequences. Some exemplary cleavage sequences may be, but are not limited to, one or more of FRAC, KRCF, KKRY, ARMA, RRSG, MRAC, KMCG, ARCA, KKQG, YRSY, SFMN, FKAA, KRNG, YNSF, KKNG, RRRG, KRYS, and ARYA (SEQ ID NOS: 62-79, respectively).

[0308] In some embodiments, the string construct is RNA (e.g., mRNA). In some embodiments, the pharmaceutical composition comprises one or more RNA (e.g., mRNA) string constructs, each comprising a sequence encoding multiple epitopes as described herein. In some embodiments, the one or more RNAs (e.g., mRNA) comprise multiple epitopes, each of which is predicted by an HLA binding and presentation prediction algorithm to have a high likelihood of being presented to T cells by an HLA-encoded protein to elicit an immune response.

[0309] In some embodiments, one or more RNAs (e.g., mRNAs) utilized in a pharmaceutical composition (e.g., an immunogenic composition, e.g., a vaccine) described herein encode multiple epitopes (e.g., including one or more, or two or more antigens provided in Table 4, Table 5, or Table 6, or fragments or epitopes thereof), optionally each of the plurality predicted by an HLA binding and presentation prediction algorithm to be likely to be presented to T cells to elicit an immune response by an HLA-encoded protein. In some embodiments, the multiple epitopes include epitopes from a single HSV (e.g., HSV-1 and / or HSV-2) protein. In some embodiments, the multiple epitopes include epitopes from multiple HSV (e.g., HSV-1 and / or HSV-2) proteins.

[0310] In some embodiments, the one or more RNAs (e.g., mRNAs) utilized in the pharmaceutical compositions (e.g., immunogenic compositions, e.g., vaccines) described herein include a first RNA encoding an HSV (e.g., HSV-1, HSV-2, or both) antigen that is expressed prior to cell invasion or infection and includes one or more fragments predicted or known to interface with the host cytoplasm. In some embodiments, the HSV antigen encoded by the first RNA is or includes an HSV antigen, fragment, or epitope, such as UL1, UL21, UL27, UL29, UL39, UL40, UL46, UL47, UL48, UL49, RS1, RL2, UL5, UL9, UL19, UL25, UL30, UL52, US1, US7, US8, UL22, and / or UL54, or fragments thereof, epitopes thereof, and / or combinations thereof. In some embodiments, one or more RNAs (e.g., mRNAs) utilized in a pharmaceutical composition (e.g., an immunogenic composition, e.g., a vaccine) described herein comprises a second antigen RNA encoding a multi-epitope (e.g., polyepitope) antigen. In some embodiments, the multi-epitope antigen comprises two or more antigens, or fragments thereof or epitopes thereof, found in Tables 3-5 herein. In some embodiments, the multi-epitope antigen comprises two or more antigens, and / or fragments and / or epitopes thereof, listed in Tables 3-5.

[0311] In some embodiments, one or more RNAs (e.g., mRNAs) utilized in a pharmaceutical composition (e.g., an immunogenic composition, e.g., a vaccine) described herein comprise multiple epitopes predicted by an HLA binding and presentation prediction algorithm to be likely to be presented to T cells by HLA-encoded proteins to elicit an immune response. In some embodiments, the multiple epitopes comprise epitopes from a single HSV (e.g., HSV-1, HSV-2, or both) protein. In some embodiments, the multiple epitopes comprise epitopes from multiple HSV (e.g., HSV-1 and / or HSV-2) proteins.

[0312] In some embodiments, the RNA (e.g., mRNA) comprises a 5' UTR and a 3' UTR. In some embodiments, the UTR comprises a polyA sequence. In some embodiments, the polyA sequence comprises between 50 and 200 nucleotides.

[0313] In some embodiments, the epitope encoded by the string construct may be flanked by signal peptide sequences, such as the SP1 sequence (HSV-1 gD signal peptide / secretory domain).

[0314] In some embodiments, the polynucleotide comprises a dEarI-hAg sequence.

[0315] In some embodiments, the poly A tail of the string construct may comprise about 150 A residues. In some embodiments, the poly A tail may comprise 120 or fewer residues. In some embodiments, the poly A tail of the string construct may comprise about 120 A residues. In some embodiments, the poly A tail of the string construct may comprise about 100 A residues. In some embodiments, the poly A tail of the string comprises a "split" or "interrupted" poly A tail (e.g., as described in WO2016 / 005324).

[0316] In some embodiments, the multi-epitope antigen encodes a supermotif- or motif-bearing polypeptide together with a helper epitope (e.g., a heterologous helper epitope) and an endoplasmic reticulum targeting signal sequence (see, e.g., An & Whitton J. Virol. 71:2292, 1997; Thomson et al., J. Immunol. 157:822, 1996; Whitton et al., J. Virol. 67:348, 1993; Hanke et al., Vaccine 16:426, 1998).

[0317] 3. T Cell Antigens and Related Constructs In certain embodiments, the HSV antigen construct can comprise and / or encode at least one T cell antigen (e.g., at least one CD4 and / or CD8 T cell antigen) or fragment thereof against HSV selected from UL1, UL21, UL27, UL29, UL39, UL40, UL46, UL47, UL48, UL49, RS1, RL2, UL5, UL9, UL19, UL25, UL30, UL52, US1, US7, US8, UL22, and / or UL54. In certain embodiments, the HSV antigen construct can comprise and / or encode at least one T cell antigen or fragment thereof against HSV selected from UL1, UL21, UL27, UL29, UL39, UL40, UL46, UL47, UL48, and / or UL49. In certain embodiments, the HSV antigen construct can comprise and / or encode at least one T cell antigen or fragment thereof against HSV selected from RS1, RL2, UL5, UL9, UL19, UL25, UL30, UL52, US1, US7, US8, UL22, and / or UL54.

[0318] In certain embodiments, the HSV antigen construct can comprise and / or encode multiple (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19) T cell antigens (e.g., CD4 and / or CD8 T cell antigens against HSV selected from UL1, UL21, UL27, UL29, UL39, UL40, UL46, UL47, UL48, UL49, RS1, RL2, UL5, UL9, UL19, UL25, UL30, UL52, US1, US7, US8, UL22, and / or UL54) or fragments thereof. In certain embodiments, the HSV antigen construct can comprise and / or encode multiple (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) T cell antigens or fragments thereof against HSV selected from UL1, UL21, UL27, UL29, UL39, UL40, UL46, UL47, UL48, and / or UL49. In certain embodiments, the HSV antigen construct can comprise and / or encode multiple (e.g., 1, 2, 3, 4, 5, 6, 7, 8, or 9) T cell antigens or fragments thereof against HSV selected from RS1, RL2, UL5, UL9, UL19, UL25, UL30, UL52, US1, US7, US8, UL22, and / or UL54.

[0319] In some embodiments, a polyribonucleotide according to the present disclosure encodes a polypeptide comprising two or more HSV antigens or antigen fragments. In some embodiments, the two or more HSV antigen fragments are each fragments of different HSV antigens. In some embodiments, at least two of the HSV antigen fragments are fragments derived from the same HSV antigen.

[0320] In some embodiments, the polyribonucleotide encodes a polypeptide, wherein the polypeptide comprises three or more HSV antigens or antigenic fragments thereof. In some embodiments, the three or more HSV antigenic fragments are each fragments of different HSV antigens. In some embodiments, at least two of the three HSV antigenic fragments are fragments from the same HSV antigen.

[0321] In some embodiments, the polyribonucleotide encodes a polypeptide, wherein the polypeptide comprises four or more HSV antigens or antigenic fragments thereof. In some embodiments, the four or more HSV antigenic fragments are each fragments of different HSV antigens. In some embodiments, at least two of the four HSV antigenic fragments are fragments from the same HSV antigen.

[0322] In some embodiments, the polyribonucleotide encodes a polypeptide, wherein the polypeptide comprises five or more HSV antigens or antigenic fragments thereof. In some embodiments, the five or more HSV antigenic fragments are each fragments of different HSV antigens. In some embodiments, at least two of the five HSV antigenic fragments are fragments from the same HSV antigen.

[0323] In some embodiments, the polyribonucleotide encodes a polypeptide, wherein the polypeptide comprises six or more HSV antigens or antigenic fragments thereof. In some embodiments, the six or more HSV antigenic fragments are each fragments of different HSV antigens. In some embodiments, at least two of the six HSV antigenic fragments are fragments from the same HSV antigen.

[0324] In some embodiments, a polypeptide according to the present disclosure does not include a full-length HSV antigen.

[0325] In some embodiments, a polypeptide according to the present disclosure comprises one or more HSV antigens or antigenic fragments thereof, including one or more T cell antigens.

[0326] In some embodiments, one or more HSV antigens or antigenic fragments thereof have at least 80% sequence identity to one or more sequences selected from SEQ ID NOs: 1-74, or their corresponding fragments.

[0327] In some embodiments, the HSV T cell antigen is or comprises a UL1 polypeptide or fragment thereof. In various embodiments, the UL1 polypeptide or fragment thereof has at least 80% sequence identity to a UL1 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. In some embodiments, the UL1 polypeptide or fragment thereof has at least 80%, e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity to a UL1 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of UL1 polypeptides known in the art include, but are not limited to, UL1 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the UL1 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NOs: 1, 2 and / or 3.

[0328] In some embodiments, the HSV T cell antigen is or comprises a UL21 polypeptide or fragment thereof. In various embodiments, the UL21 polypeptide or fragment thereof has at least 80% sequence identity to a UL21 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. In some embodiments, the UL21 polypeptide or fragment thereof has at least 80%, e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity to a UL1 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of UL21 polypeptides known in the art include, but are not limited to, UL21 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the UL21 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NOs: 4, 5 and / or 6.

[0329] The UL27 open reading frame encodes HSV gB (also referred to herein as the UL27 polypeptide). In some embodiments, the HSV T cell antigen is or comprises a UL27 polypeptide or a fragment thereof. In various embodiments, the UL27 polypeptide or fragment thereof has at least 80% sequence identity to a UL27 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. In some embodiments, the UL27 polypeptide or fragment thereof has at least 80%, e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity to a UL1 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of UL27 polypeptides known in the art include, but are not limited to, UL27 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the UL27 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NOs: 7, 8, 9 and / or 74.

[0330] In some embodiments, the HSV T cell antigen is or comprises a UL29 polypeptide or fragment thereof. In various embodiments, the UL29 polypeptide or fragment thereof has at least 80% sequence identity to a UL29 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. In some embodiments, the UL29 polypeptide or fragment thereof has at least 80%, e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity to a UL1 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of UL29 polypeptides known in the art include, but are not limited to, UL29 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the UL29 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NOs: 10, 11 and / or 12.

[0331] In some embodiments, the HSV T cell antigen is or comprises a UL39 polypeptide or fragment thereof. In various embodiments, the UL39 polypeptide or fragment thereof has at least 80% sequence identity to a UL39 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. In some embodiments, the UL39 polypeptide or fragment thereof has at least 80%, e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity to a UL1 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of UL39 polypeptides known in the art include, but are not limited to, UL39 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the UL39 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NOs: 13, 14, and / or 15.

[0332] In some embodiments, the HSV T cell antigen is or comprises a UL40 polypeptide or fragment thereof. In various embodiments, the UL40 polypeptide or fragment thereof has at least 80% sequence identity to a UL40 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. In some embodiments, the UL1 polypeptide or fragment thereof has at least 80%, e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity to a UL40 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of UL40 polypeptides known in the art include, but are not limited to, UL40 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the UL40 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NOs: 16, 17 and / or 18.

[0333] In some embodiments, the HSV T cell antigen is or comprises a UL46 polypeptide or fragment thereof. In various embodiments, the UL46 polypeptide or fragment thereof has at least 80% sequence identity to a UL46 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. In some embodiments, the UL46 polypeptide or fragment thereof has at least 80%, e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity to a UL1 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of UL46 polypeptides known in the art include, but are not limited to, UL46 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the UL46 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NOs: 19, 20 and / or 21.

[0334] In some embodiments, the HSV T cell antigen is or comprises a UL47 polypeptide or fragment thereof. In various embodiments, the UL47 polypeptide or fragment thereof has at least 80% sequence identity to a UL47 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. In some embodiments, the UL47 polypeptide or fragment thereof has at least 80%, e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity to a UL1 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of UL47 polypeptides known in the art include, but are not limited to, UL47 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the UL47 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NOs: 22, 23 and / or 24.

[0335] In some embodiments, the HSV T cell antigen is or comprises a UL48 polypeptide or fragment thereof. In various embodiments, the UL48 polypeptide or fragment thereof has at least 80% sequence identity to a UL48 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. In some embodiments, the UL48 polypeptide or fragment thereof has at least 80%, e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity to a UL1 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of UL48 polypeptides known in the art include, but are not limited to, UL48 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the UL48 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NOs: 25, 26 and / or 27.

[0336] In some embodiments, the HSV T cell antigen is or comprises a UL49 polypeptide or fragment thereof. In various embodiments, the UL49 polypeptide or fragment thereof has at least 80% sequence identity to a UL49 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. In some embodiments, the UL49 polypeptide or fragment thereof has at least 80%, e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity to a UL1 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of UL49 polypeptides known in the art include, but are not limited to, UL49 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the UL49 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NOs: 28, 29, and / or 30.

[0337] In some embodiments, the HSV T cell antigen is or comprises an RS1 polypeptide or a fragment thereof. In various embodiments, the RS1 polypeptide or fragment thereof has at least 80% sequence identity to an RS1 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. In some embodiments, the UL1 polypeptide or fragment thereof has at least 80%, e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity to an UL1 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of RS1 polypeptides known in the art include, but are not limited to, RS1 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the RS1 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NOs: 31, 32, and / or 33.

[0338] In some embodiments, the HSV T cell antigen is or comprises an RL2 polypeptide or a fragment thereof. In various embodiments, the RL2 polypeptide or fragment thereof has at least 80% sequence identity to the RL2 amino acid sequence shown in Table 3 or otherwise known in the art, or a corresponding fragment. In some embodiments, the RL2 polypeptide or fragment thereof has at least 80%, e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity to the UL1 amino acid sequence shown in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of RL2 polypeptides known in the art include, but are not limited to, RL2 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the RL2 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NOs: 34, 35, and / or 36.

[0339] In some embodiments, the HSV T cell antigen is or comprises a UL5 polypeptide or fragment thereof. In various embodiments, the UL5 polypeptide or fragment thereof has at least 80% sequence identity to a UL5 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. In some embodiments, the UL5 polypeptide or fragment thereof has at least 80%, e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity to a UL1 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of UL5 polypeptides known in the art include, but are not limited to, UL5 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the UL5 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NOs: 37, 38 and / or 39.

[0340] In some embodiments, the HSV T cell antigen is or comprises a UL9 polypeptide or fragment thereof. In various embodiments, the UL9 polypeptide or fragment thereof has at least 80% sequence identity to a UL9 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. In some embodiments, the UL9 polypeptide or fragment thereof has at least 80%, e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity to a UL1 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of UL9 polypeptides known in the art include, but are not limited to, UL9 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the UL9 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NO: 40, 41 and / or 42.

[0341] In some embodiments, the HSV T cell antigen is or comprises a UL19 polypeptide or fragment thereof. In various embodiments, the UL19 polypeptide or fragment thereof has at least 80% sequence identity to a UL19 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. In some embodiments, the UL19 polypeptide or fragment thereof has at least 80%, e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity to a UL1 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of UL19 polypeptides known in the art include, but are not limited to, UL19 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the UL19 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NO: 43, 44, and / or 45.

[0342] In some embodiments, the HSV T cell antigen is or comprises a UL25 polypeptide or fragment thereof. In various embodiments, the UL25 polypeptide or fragment thereof has at least 80% sequence identity to a UL25 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. In some embodiments, the UL25 polypeptide or fragment thereof has at least 80%, e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity to a UL1 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of UL25 polypeptides known in the art include, but are not limited to, UL25 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the UL25 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NOs: 46, 47, and / or 48.

[0343] In some embodiments, the HSV T cell antigen is or comprises a UL30 polypeptide or fragment thereof. In various embodiments, the UL30 polypeptide or fragment thereof has at least 80% sequence identity to a UL30 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. In some embodiments, the UL30 polypeptide or fragment thereof has at least 80%, e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity to a UL1 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of UL30 polypeptides known in the art include, but are not limited to, UL30 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the UL30 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NOs: 49, 50, and / or 51.

[0344] In some embodiments, the HSV T cell antigen is or comprises a UL52 polypeptide or fragment thereof. In various embodiments, the UL52 polypeptide or fragment thereof has at least 80% sequence identity to a UL52 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. In some embodiments, the UL52 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a UL1 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of UL52 polypeptides known in the art include, but are not limited to, UL52 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the UL52 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NO: 52, 53 and / or 54.

[0345] The US1 open reading frame encodes HSV gL (also referred to herein as the UL1 polypeptide). In some embodiments, the HSV T cell antigen is or comprises a US1 polypeptide or a fragment thereof. In various embodiments, the US1 polypeptide or fragment thereof has at least 80% sequence identity to a US1 amino acid sequence set forth in Table 3 or otherwise known in the art, or its corresponding fragment. In some embodiments, the Us1 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a UL1 amino acid sequence set forth in Table 3 or otherwise known in the art, or its corresponding fragment. Examples of US1 polypeptides known in the art include, but are not limited to, US1 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the US1 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NOs: 58, 59, 60, and / or 61.

[0346] The US7 open reading frame encodes an HSV gI (also referred to herein as a US7 polypeptide). In some embodiments, the HSV T cell antigen is or comprises a US7 polypeptide or a fragment thereof. In various embodiments, the US7 polypeptide or fragment thereof has at least 80% sequence identity to a US7 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. In some embodiments, the UL1 polypeptide or fragment thereof has at least 80%, e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, or 100% sequence identity to a US7 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of US7 polypeptides known in the art include, but are not limited to, US7 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the US7 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NOs: 62, 63, 64, and / or 65.

[0347] The US8 open reading frame encodes HSV gE (also referred to herein as the US8 polypeptide). In some embodiments, the HSV T cell antigen is or comprises a US8 polypeptide or a fragment thereof. In various embodiments, the US8 polypeptide or fragment thereof has at least 80% sequence identity to a US8 amino acid sequence set forth in Table 3 or otherwise known in the art, or its corresponding fragment. In some embodiments, the UL1 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a US8 amino acid sequence set forth in Table 3 or otherwise known in the art, or its corresponding fragment. Examples of US8 polypeptides known in the art include, but are not limited to, US8 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the US8 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NOs: 66, 67, 68, and / or 69.

[0348] The UL22 open reading frame encodes an HSV gH (also referred to herein as a UL22 polypeptide). In some embodiments, the HSV T cell antigen is or comprises a UL22 polypeptide or a fragment thereof. In various embodiments, the UL22 polypeptide or fragment thereof has at least 80% sequence identity to a UL22 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. In some embodiments, the UL22 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a UL1 amino acid sequence set forth in Table 3 or otherwise known in the art, or a corresponding fragment thereof. Examples of UL22 polypeptides known in the art include, but are not limited to, UL22 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the UL22 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NOs: 70, 71, 72 and / or 73.

[0349] In some embodiments, the HSV T cell antigen is or comprises a UL54 polypeptide or fragment thereof. In various embodiments, the UL54 polypeptide or fragment thereof has at least 80% sequence identity to a UL54 amino acid sequence set forth in Table 3 or otherwise known in the art, or its corresponding fragment. In some embodiments, the UL1 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a UL54 amino acid sequence set forth in Table 3 or otherwise known in the art, or its corresponding fragment. Examples of UL54 polypeptides known in the art include, but are not limited to, UL54 polypeptides encoded by known HSV strains, such as HG52, G, 333, and MS strains. In some embodiments, the UL54 polypeptide or fragment thereof has at least 80%, e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence set forth in SEQ ID NO: 55, 56 and / or 57.

[0350] In some embodiments, an HSV antigen for use in accordance with the present disclosure comprises an RL2 polypeptide or an antigenic fragment thereof, an RS1 polypeptide or an antigenic fragment thereof, a UL54 polypeptide or an antigenic fragment thereof, a UL29 polypeptide or an antigenic fragment thereof, a UL39 polypeptide or an antigenic fragment thereof, a UL49 polypeptide or an antigenic fragment thereof, a UL9 polypeptide or an antigenic fragment thereof, a UL30 polypeptide or an antigenic fragment thereof, a UL40 polypeptide or an antigenic fragment thereof, a UL5 polypeptide or an antigenic fragment thereof, a UL52 polypeptide or an antigenic fragment thereof, a UL1 polypeptide or an antigenic fragment thereof, a UL19 polypeptide or an antigenic fragment thereof, a UL21 polypeptide or an antigenic fragment thereof, a UL27 polypeptide or an antigenic fragment thereof, a UL46 polypeptide or an antigenic fragment thereof, a UL47 polypeptide or an antigenic fragment thereof, a UL48 polypeptide or an antigenic fragment thereof, a UL25 polypeptide or an antigenic fragment thereof, or a combination thereof.

[0351] In some embodiments, the polyribonucleotides provided herein encode one or more of an RL2 polypeptide or antigenic fragment thereof, an RS1 polypeptide or antigenic fragment thereof, a UL54 polypeptide or antigenic fragment thereof, a UL29 polypeptide or antigenic fragment thereof, a UL39 polypeptide or antigenic fragment thereof, a UL49 polypeptide or antigenic fragment thereof, a UL9 polypeptide or antigenic fragment thereof, a UL30 polypeptide or antigenic fragment thereof, a UL40 polypeptide or antigenic fragment thereof, a UL5 polypeptide or antigenic fragment thereof, a UL52 polypeptide or antigenic fragment thereof, a UL1 polypeptide or antigenic fragment thereof, a UL19 polypeptide or antigenic fragment thereof, a UL21 polypeptide or antigenic fragment thereof, a UL27 polypeptide or antigenic fragment thereof, a UL46 polypeptide or antigenic fragment thereof, a UL47 polypeptide or antigenic fragment thereof, a UL48 polypeptide or antigenic fragment thereof, a UL25 polypeptide or antigenic fragment thereof, a US1 polypeptide or antigenic fragment thereof, a US7 polypeptide or antigenic fragment thereof, a US8 polypeptide or antigenic fragment thereof, and a UL22 polypeptide or antigenic fragment thereof.

[0352] In some embodiments, the polyribonucleotides provided herein encode one or more HSV (e.g., HSV-1 and / or HSV-2) antigens, wherein at least one antigen is an RL2 polypeptide or antigenic fragment thereof. In some embodiments, the RL2 polypeptide or antigenic fragment thereof comprises or consists of the following amino acid sequence: CTDEIAPPLRCQSFPCLHPFCIPCMKTWIPLRNTCPLCNTPVAYLIVGVTASGSFSTIPIVNDPRTRVEAEAAVRAGTAVDFIWTGNPRTAPRSLS (SEQ ID NO: 174). In some embodiments, the RL2 polypeptide or antigenic fragment thereof is It comprises or consists of the amino acid sequence of LPIAGVSSVVALAPYVNKTVTGDCLPVLDMETGHIGAYVVLVDQTGNVADLLRAAAPAWSRRTLLPEHARNCVRPPDYPTPPASEWNSLWMTPVGNMLFDQGTLVG (SEQ ID NO: 175).

[0353] In some embodiments, the polyribonucleotides provided herein encode one or more HSV (e.g., HSV-1 and / or HSV-2) antigens, wherein at least one antigen is an RS1 polypeptide or antigenic fragment thereof. In some embodiments, the RS1 polypeptide or antigenic fragment thereof comprises or consists of the following amino acid sequence: RAAAWMRQVPDPEDVRVVILYSPLPGEDLAAGRAGGGPPPEWSAERGGLSCLLAALGNRLCGPATAAWAGNWTGAPDVSALGAQGVLLLSTRDLAFAGAVEFLGLLAGACDRRLIVVNAVRAADWPADGPVVSRQHAYLACEVLPAVQCAVRWPAARDLRRTVLASGRVFGPGVFARVEAAHARLYPDAPPLRLCRGANVRYRVRTRFGPDTLVPMSPREYRRAVLPALDGRAAAS (SEQ ID NO: 176).

[0354] In some embodiments, the polyribonucleotides provided herein encode one or more HSV (e.g., HSV-1 and / or HSV-2) antigens, wherein at least one antigen is a UL54 polypeptide or antigenic fragment thereof. In some embodiments, the UL54 polypeptide or antigenic fragment thereof comprises or consists of the following amino acid sequence: ETLVAHGPSLYRTFAANPRAASTAKAMRDCVLRQENLIEALASADETLAWCKMCIHHNLPLRPQDPIIGTAAAVLENLATRLRPFLQCYLKARGLCGLDDLCSRRRLSDIKDIASFVLVILARLANRVERGVSEIDYTTVGVGAGETMHFYIPGACMAGLIEILDTHRQECSSRVCELTASHTIAPLYVHGKYFYCNSLF (SEQ ID NO: 177).

[0355] In some embodiments, the polyribonucleotides provided herein encode one or more HSV (e.g., HSV-1 and / or HSV-2) antigens, wherein at least one antigen is a UL29 polypeptide or antigenic fragment thereof. In some embodiments, the UL29 polypeptide or antigenic fragment thereof comprises or consists of the following amino acid sequence: REDIETIAFIKRFSLDYGAINFINLAPNNVSELAMYYMANQILRYCDHSTYFINTLTAVIAGSRRPPSVQAAAAWAPQGGAGLEAGARALMDSLDAHPGAWTSMFASCNLLRPVMAARPMVVLGLSISKYYGMAGNDRVFQAGNWASLLGGKNACPLLIFDRTRKFVL (SEQ ID NO: 178).

[0356] In some embodiments, the polyribonucleotides provided herein encode one or more HSV (e.g., HSV-1 and / or HSV-2) antigens, wherein at least one antigen is a UL39 polypeptide or antigenic fragment thereof. In some embodiments, the UL39 polypeptide or antigenic fragment thereof comprises or consists of the following amino acid sequence: RTFGSAPRLTEDDFGLLNYALAEMRRLCLDLPPVPPNAYTPYHLREYATRLVNGFKPLVRRSARLYRILGVLVHLRIRTREASFEEWMRSKEVDLDFGLTERLREHEAQLMILAQALNPYDCLIHSTPNTLVERGLQSALKYEEFYLKRFGGHYMESVFQMYTRIAGFLA (SEQ ID NO: 179).

[0357] In some embodiments, the polyribonucleotides provided herein encode one or more HSV (e.g., HSV-1 and / or HSV-2) antigens, wherein at least one antigen is a UL49 polypeptide or antigenic fragment thereof. In some embodiments, the UL49 polypeptide or antigenic fragment thereof comprises or consists of the following amino acid sequence: KMTRGAPKASATPATDPARGRRPAQADSAVLLDAPAPTASGRTKTPAQGLAKKLHFSTAPPSPTAPWTPRVAGFNKRVFCAAVG (SEQ ID NO: 180).

[0358] In some embodiments, the polyribonucleotides provided herein encode one or more HSV (e.g., HSV-1 and / or HSV-2) antigens, wherein at least one antigen is a UL9 polypeptide or antigenic fragment thereof. In some embodiments, the UL9 polypeptide or antigenic fragment thereof comprises or consists of the following amino acid sequence: LLNNYDVLVLDEVMSTLGQLYSPTMQQLGRVDALMLRLLRTCPRIIAMDATANAQLVDFLCSLRGEKNVHVVIGEYAMPGFSARRCLFLPRLGPEVLQAALRRRGPAGGAPPPDAPPDATFFGELEARLAGGDNVCIFSSTVSFAEVVARFCRQFTDRVLLLHSLTPPGDVTTWGRYRVVIYTTVVTVGLSFDPPHFDSMFAYVKPMNYGPDMVSVYQSLGRVRTLRKGELLIYMDGSGARSEPV (SEQ ID NO: 181).

[0359] In some embodiments, the polyribonucleotides provided herein encode one or more HSV (e.g., HSV-1 and / or HSV-2) antigens, wherein at least one antigen is a UL30 polypeptide or antigenic fragment thereof. In some embodiments, the UL30 polypeptide or antigenic fragment thereof comprises or consists of the following amino acid sequence: ISCLLYDLSTTALEHILLFSLGSCDLPESHLSDLASRGLPAPVVLEFDSEFEMLLAFMTFVKQYGPEFVTGYNIINFDWPFVLTKLTEIYKVPLDGYGRMNGRGVFRVWDIGQSHF (SEQ ID NO: 182). In some embodiments, the UL30 polypeptide or antigenic fragment thereof is It contains the amino acid sequence of GLLPCLHVAATVTTIGREMLLATRAYVHARWAEFDQLLADFPEAAGMRAPGPYSM (SEQ ID NO: 183).

[0360] In some embodiments, the polyribonucleotides provided herein encode one or more HSV (e.g., HSV-1 and / or HSV-2) antigens, wherein at least one antigen is a UL40 polypeptide or antigenic fragment thereof. In some embodiments, the UL40 polypeptide or antigenic fragment thereof comprises or consists of the following amino acid sequence: TSQCPDINHLRSLSILNRWLETELVFVGDEEDVSKLSEGELGFYRFLFAFLSAADDLVTENLGGLSGLFEQKDILHYYVEQECIEVVHSRVYNIIQLVLFHNNDQARRAYVARTINHPAIRVKVDWLEARVRECDSIPEKFILMILIEGVFFAASFAAIAYLRTNNLLR (SEQ ID NO: 184).

[0361] In some embodiments, the polyribonucleotides provided herein encode one or more HSV (e.g., HSV-1 and / or HSV-2) antigens, wherein at least one antigen is a UL5 polypeptide or antigenic fragment thereof. In some embodiments, the UL5 polypeptide or antigenic fragment thereof comprises or consists of the following amino acid sequence: HEFGNLMKVLEYGLPITEEHMQFVDRFVVPESYITNPANLPGWTRLFSSHKEVSAYMAKLHAYLKVTREGEFVVFTLPVLTFVSVKEFDEYRRL (SEQ ID NO: 185). In some embodiments, the UL5 polypeptide or antigenic fragment thereof comprises or consists of the following amino acid sequence: ELFGEVFESAPFSTYVDNVIFRGCELLTGSPRGGLMSVALQTDNYTLMGYTYTRVFAFAEELRRRHATAGVAEFLEESPLPYIVLRDQHGFMSVVNTNI (SEQ ID NO: 186).

[0362] In some embodiments, the polyribonucleotides provided herein encode one or more HSV (e.g., HSV-1 and / or HSV-2) antigens, wherein at least one antigen is a UL52 polypeptide or antigenic fragment thereof. In some embodiments, the UL52 polypeptide or antigenic fragment thereof comprises or consists of the following amino acid sequence: SVAAPVEVTALYATDGCVITSSLALLTNCLLGAEPLYIFSYDAYRSDAPNGPTGAPTEQERFEGSRALYRDAGGLNGDSFRVTFCLLGTEVGVTHHPKGRTRPMFVCRFERADDVAVLQDALGRGTPLLPAHVTATLDLEATFALHANIIMALTVAIVHNAPARIGSGSTAPLYEPGESMRSVV (SEQ ID NO: 187).

[0363] In some embodiments, the polyribonucleotides provided herein encode one or more HSV (e.g., HSV-1 and / or HSV-2) antigens, wherein at least one antigen is a UL1 polypeptide or antigenic fragment thereof. In some embodiments, the UL1 polypeptide or antigenic fragment thereof comprises or consists of the following amino acid sequence: RTPADDVSWRYEAPSVIDYARIDGIFLRYHCPGLDTFLWDRHAQRAYLVNPFLFAAGFLEDLSHSVFPADTQETT (SEQ ID NO: 188).

[0364] In some embodiments, the polyribonucleotides provided herein encode one or more HSV (e.g., HSV-1 and / or HSV-2) antigens, wherein at least one antigen is a UL19 polypeptide or antigenic fragment thereof. In some embodiments, the UL19 polypeptide or antigenic fragment thereof comprises or consists of the following amino acid sequence: DGRLLHNTQARAADAADDRPHRPADWTVHHKIYYYVLVPAFSRGRCCTAGVRFDRVYATLQNMVVPEIAPGEECPSDPVTDPAHPLHPANLVANTVKRMFHN (SEQ ID NO: 189).

[0365] In some embodiments, the polyribonucleotides provided herein encode one or more HSV (e.g., HSV-1 and / or HSV-2) antigens, wherein at least one antigen is a UL21 polypeptide or antigenic fragment thereof. In some embodiments, the UL21 polypeptide or antigenic fragment thereof comprises or consists of the following amino acid sequence: SPTQKLAVYYYLIHRERRMSPFPALVRLVGRYIQRHGLYVPAPDEPTLADAMNGL (SEQ ID NO: 190).

[0366] In some embodiments, the polyribonucleotides provided herein encode one or more HSV (e.g., HSV-1 and / or HSV-2) antigens, wherein at least one antigen is a UL27 polypeptide or antigenic fragment thereof. In some embodiments, the UL27 polypeptide or antigenic fragment thereof comprises or consists of the following amino acid sequence: NYTEGIAVVFKENIAPYKFKATMYYKDVTVSQVWFGHRYSQFMGIFEDRAPVPFEEV (SEQ ID NO: 191). In some embodiments, the UL27 polypeptide or antigenic fragment thereof is It comprises or consists of the amino acid sequence SVYPYDEFVLATGDFVYMSPFYGYREGSH (SEQ ID NO: 192).

[0367] In some embodiments, the polyribonucleotides provided herein encode one or more HSV (e.g., HSV-1 and / or HSV-2) antigens, wherein at least one antigen is a UL46 polypept...

Claims

1. 1. A polyribonucleotide encoding a polypeptide, said polypeptide comprising two or more herpes simplex virus (HSV) antigens or antigenic fragments thereof, said two or more HSV antigens or antigenic fragments comprising: (i) one or more HSV RS1 polypeptides or antigenic fragments thereof; (ii) one or more HSV RL2 polypeptides or antigenic fragments thereof; (iii) one or more HSV UL1 polypeptides or antigenic fragments thereof; (iv) one or more HSV UL5 polypeptides or antigenic fragments thereof; (v) one or more HSV UL9 polypeptides or antigenic fragments thereof; (vi) one or more HSV UL19 polypeptides or antigenic fragments thereof; (vii) one or more HSV UL21 polypeptides or antigenic fragments thereof; (viii) one or more HSV UL25 polypeptides or antigenic fragments thereof; (ix) one or more HSV UL27 polypeptides or antigenic fragments thereof; (x) one or more HSV UL29 polypeptides or antigenic fragments thereof; (xi) one or more HSV UL30 polypeptides or antigenic fragments thereof; (xii) one or more HSV UL39 polypeptides or antigenic fragments thereof; (xiii) one or more HSV UL40 polypeptides or antigenic fragments thereof; (xiv) one or more HSV UL46 polypeptides or antigenic fragments thereof; (xv) one or more HSV UL47 polypeptides or antigenic fragments thereof; (xvi) one or more HSV UL48 polypeptides or antigenic fragments thereof; (xvii) one or more HSV UL49 polypeptides or antigenic fragments thereof; (xviii) one or more HSV UL52 polypeptides or antigenic fragments thereof, and (xix) one or more HSV UL54 polypeptides or antigenic fragments thereof wherein the two or more HSV antigen fragments are each fragments of different HSV antigens.

2. The polyribonucleotide of claim 1, wherein the polypeptide comprises an HSV-1 gD secretion signal.

3. The polyribonucleotide described in claim 1, wherein the polypeptide comprises MITD.

4. A polyribonucleotide as described in claim 1, wherein the polypeptide comprises one or more linkers.

5. 2. The polyribonucleotide of claim 1, wherein the polypeptide comprises one or more HSV RL2 polypeptides or antigenic fragments thereof, one or more HSV RS1 polypeptides or antigenic fragments thereof, and one or more HSV UL54 polypeptides or antigenic fragments thereof.

6. 2. The polyribonucleotide of claim 1, wherein the polypeptide comprises one or more HSV UL29 polypeptides or antigenic fragments thereof, one or more HSV UL39 polypeptides or antigenic fragments thereof, one or more HSV UL49 polypeptides or antigenic fragments thereof, and one or more HSV UL9 polypeptides or antigenic fragments thereof.

7. 2. The polyribonucleotide of claim 1, wherein the polypeptide comprises one or more HSV UL30 polypeptides or antigenic fragments thereof, one or more HSV UL40 polypeptides or antigenic fragments thereof, one or more HSV UL5 polypeptides or antigenic fragments thereof, and one or more HSV UL52 polypeptides or antigenic fragments thereof.

8. 2. The polyribonucleotide of claim 1, wherein the polypeptide comprises one or more HSV UL1 polypeptides or antigenic fragments thereof, one or more HSV UL19 polypeptides or antigenic fragments thereof, one or more HSV UL21 polypeptides or antigenic fragments thereof, one or more HSV UL27 polypeptides or antigenic fragments thereof, one or more HSV UL46 polypeptides or antigenic fragments thereof, one or more HSV UL47 polypeptides or antigenic fragments thereof, one or more UL48 polypeptides or antigenic fragments thereof, and one or more HSV UL25 polypeptides or antigenic fragments thereof.

9. The polyribonucleotide of claim 1, wherein the polyribonucleotide is a codon-optimized polyribonucleotide.

10. The two or more selected HSV antigens: (i) one or more HSV RL2 polypeptides or antigenic fragments thereof; (ii) one or more HSV UL54 polypeptides or antigenic fragments thereof; (iii) one or more HSV UL5 polypeptides or antigenic fragments thereof; (iv) one or more HSV UL40 polypeptides or antigenic fragments thereof; (v) one or more HSV UL47 polypeptides or antigenic fragments thereof; (vi) one or more HSV UL46 polypeptides or antigenic fragments thereof, or (vii) any combination thereof The polyribonucleotide of claim 1, comprising:

11. The two or more selected HSV antigens: (i) one or more HSV RL2 polypeptides or antigenic fragments thereof, and one or more HSV UL54 polypeptides or antigenic fragments thereof; (ii) one or more HSV UL5 polypeptides or antigenic fragments thereof and one or more HSV UL40 polypeptides or antigenic fragments thereof; or (iii) one or more HSV UL47 polypeptides or antigenic fragments thereof, and one or more HSV UL46 polypeptides or antigenic fragments thereof. The polyribonucleotide of claim 1, comprising:

12. The two or more selected HSV antigens: one or more HSV RL2 polypeptides or antigenic fragments thereof, one or more HSV UL54 polypeptides or antigenic fragments thereof, one or more HSV UL40 polypeptides or antigenic fragments thereof, one or more HSV UL47 polypeptides or antigenic fragments thereof, and / or one or more HSV UL46 polypeptides or antigenic fragments thereof. The polyribonucleotide of claim 1, comprising:

13. A first polyribonucleotide according to claim 1; a second polyribonucleotide, wherein the second polyribonucleotide encodes a second polypeptide, the second polypeptide comprising one or more herpes simplex virus (HSV) antigens or antigenic fragments thereof; the first polyribonucleotide and the second polyribonucleotide are different; A combination wherein said one or more HSV antigens or antigenic fragments thereof of said second polypeptide comprise one or more HSV glycoproteins.

14. 14. The combination of claim 13, wherein the one or more HSV glycoproteins comprise HSV glycoprotein B (gB), HSV glycoprotein E (gE), HSV glycoprotein G (gG), HSV glycoprotein H (gH), HSV glycoprotein I (gI), HSV glycoprotein L (gL), or a combination thereof.

15. (i) a 5′ UTR; (ii) a polyribonucleotide according to claim 1; (iii) a 3′UTR, and (iv) comprising a poly A tail sequence in 5' to 3' order; The RNA construct optionally further comprising a 5' cap.

16. 1. A composition comprising: (i) one or more polyribonucleotides according to claim 1; (ii) a lipid nanoparticle, polyplex (PLX), or liposome; The composition, wherein the one or more polyribonucleotides are fully or partially encapsulated within the lipid nanoparticle, the polyplex (PLX), or the liposome.

17. A pharmaceutical composition comprising one or more polyribonucleotides according to claim 1 and at least one pharmaceutically acceptable excipient.

18. A composition comprising a polyribonucleotide according to any one of claims 1 to 12, or an RNA construct according to claim 15, a combination according to claim 13 or 14, a composition according to claim 16, or a pharmaceutical composition according to claim 17, for use in the treatment or prevention of HSV.

19. A composition comprising the first polyribonucleotide of claim 1, The composition is used in combination with a second polyribonucleotide, wherein the second polyribonucleotide encodes a second polypeptide, and the second polypeptide comprises one or more herpes simplex virus (HSV) glycoproteins or fragments thereof; The composition, wherein said first polyribonucleotide and said second polyribonucleotide are different.

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