Combination vaccines and methods of use

Abstract

The present disclosure provides immunogenic compositions comprising RSV and VZV antigens, as well as their use in the prevention and / or treatment of VZV and RSV infections.

Description

COMBINATION VACCINES AND METHODS OF USE CROSS-REFERENCE TORELATEDAPPLICATIONS

[0001] The present application claims priority to U.S. Provisional Application 63 / 617,367, filed January 3, 2024, the contents of which are incorporated herein by reference in its entirety. REFERENCE TO ANELECTRONICSEQUENCELISTING

[0002] The contents of the electronic sequence listing (CURV_006_01WO_SeqList_ST26.xml; Size: 8,628 bytes; and Date of Creation: December 30, 2024) are herein incorporated by reference in its entirety. FIELD

[0003] The present disclosure relates generally to immunogenic compositions comprising RSV and VZV antigens, as well as the prevention and / or treatment of VZV and RSV infections and reactivation. BACKGROUND

[0004] Varicella Zoster Virus (VZV), also known as human herpesvirus 3 (HHV-3, HHV3) or Human alphaherpesvirus 3 (taxonomically), is one of nine known herpes viruses that can infect humans. It causes chickenpox (varicella – primary infection) commonly affecting children and young adults, and shingles (herpes zoster – reactivation of latent VZV) in adults and immunocompromised adolescents and children. VZV infections are species- specific to humans.

[0005] Similar to the herpes simplex viruses, after primary infection with VZV (chickenpox), the virus lies dormant in neurons, including the cranial nerve ganglia, dorsal root ganglia, and autonomic ganglia. Many years after the person has recovered from initial chickenpox infection, VZV can reactivate to cause shingles.

[0006] Respiratory Syncytial Virus (RSV), also called human respiratory syncytial virus (hRSV) and human orthopneumovirus, is a common, contagious virus that causes infections of the upper and lower respiratory tract. It is a negative-sense, single-stranded RNA virus. RSV is a common cause of respiratory hospitalization in infants, and reinfection remains common in life though often with less severity. It is a notable pathogen in all age groups. In the northern and southern hemispheres, infection rates are typically higher during the cold 1 311398121winter months. In tropical and semitropical climates, the seasonal οսtbreаkѕ usually are associated with the rainy season. Almost all сhildren are infected by two years of age. The clinical manifestations vary with age, health status, and whether the infection is primary or secondary. RSV is the most common cause of bronchiolitis in children. In adults, the clinical presentation of RSV varies and can range from сold-like symptoms to acute respiratory distress. Although asymptomatic infection can occur, it is rare in adults or the elderly (<5 percent). Patients with multiple comorbidities (i.e., immunocompromising conditions, chronic obstructive pulmonary disease [COPD] and congestive heart failure [CHF]) are more likely to develop symptomatic RSV illness.

[0007] There is a need in the art for compositions and methods suitable for the treatment and prevention of both VZV and RSV infections. SUMMARY

[0008] In some embodiments, the present disclosure provides an immunogenic composition comprising: a Varicella Zoster Virus (VZV) gE glycoprotein comprising the amino acid sequence of SEQ ID NO: 1 or a polynucleotide encoding the same; a Respiratory Syncytial Virus (RSV) antigen or a polynucleotide encoding the same; and an adjuvant.

[0009] In some embodiments, the RSV antigen comprises a pre-fusion form of one or more RSV glycoproteins. In some embodiments, the one or more RSV antigens comprise the F glycoprotein. In some embodiments, the RSV glycoprotein F comprises the amino acid sequence of any one of SEQ ID NOs: 2-4.

[0010] In some embodiments, the immunogenic composition comprises between 25 µg and 125 µg of the VZV gE antigen. In some embodiments, the immunogenic composition comprises about 100 µg of the VZV gE antigen. In some embodiments, the immunogenic composition comprises between 60 µg and 180 µg of the RSV antigen. In some embodiments, the immunogenic composition comprises about 120 µg of the RSV antigen. In some embodiments, the RSV antigen is the prefusion form of glycoprotein F. In some embodiments, the immunogenic composition comprises about 60 µg each of two RSV antigens. In some embodiments, the two RSV antigens are the prefusion forms of glycoprotein F from the A2 and B1 strains (SEQ ID NOs: 3 and 4).

[0011] In some embodiments, the adjuvant is selected from aluminum hydroxide, aluminum phosphate, aluminum hydrophosphate sulfate, MPL, QS-21, MF59C.1, CpG 1018, Matrix-M, MF59, PF-03512676, Matrix-M-2, Vaxfectin, GLA-SE, SLA-SE, 3M-052-SE, 2 311398121GLA-LSQ, SLA-LSQ, 3M-052-Alum, GLA-AF, SLA-AF, 3M-052-AF, GLA-3M-052-LS, NanoAlum, a nanostructured lipid carrier (NLC), or a combination thereof. In some embodiments, the adjuvant is SLA-SE.

[0012] In some embodiments, the present disclosure provides a method of stimulating an immune response against Varicella Zoster Virus (VZV) and / or Respiratory Syncytial Virus (RSV) in a subject comprising administering an immunogenic composition described herein.

[0013] In some embodiments, the present disclosure provides a method of preventing infection with Varicella Zoster Virus (VZV) and / or Respiratory Syncytial Virus (RSV) in a subject comprising administering an immunogenic composition described herein.

[0014] In some embodiments, a single dose of the immunogenic composition is administered. In some embodiments, the method further comprises administering a second immunogenic composition comprising the VZV gE antigen and an adjuvant. In some embodiments, the second immunogenic composition is administered 2-6 months after administration of the first immunogenic composition. In some embodiments, two or more doses of the immunogenic composition are administered. In some embodiments, the second dose is administered 2-6 months after the first dose. DETAILED DESCRIPTION

[0015] Unless defined otherwise, all technical and scientific terms used herein have the meaning commonly understood by one of ordinary skill in the art to which the invention pertains. Specific terminology of particular importance to the description of the present invention is defined below. In this specification and the appended claims, the singular forms “a,” “an” and “the” include plural referents unless the context clearly dictates otherwise. Thus, for example, “a polypeptide” refers not only to a single polypeptide but also to a combination of two or more different polypeptides that may or may not be combined, “an adjuvant” refers to a single adjuvant as well as to two or more adjuvants that may be separate or combined in a single composition, and the like.

[0016] The terms “about” and “approximately” when referring to a measurable value such as an amount, a temporal duration, and the like, are meant to encompass variations of ±20% or in some instances ±10%, or in some instances ±5%, or in some instances ±1%, or in some instances ±0.1% from the specified value, as such variations are appropriate to perform the disclosed methods. 3 311398121VZV and RSV Antigens

[0017] In some embodiments, the present disclosure provides immunogenic compositions comprising VZV and RSV polypeptide antigens. As used herein, the term “antigen” refers to a substance such as a polypeptide or peptide that is capable of eliciting an immune response.

[0018] The term “polypeptide” is intended to include any structure comprised of one or more amino acids, and thus includes dipeptides, oligopeptides, polypeptides, polypeptide fragments, and proteins. The amino acids forming all or a part of a polypeptide may be any of the twenty conventional, naturally occurring amino acids, i.e., alanine (A), cysteine (C), aspartic acid (D), glutamic acid (E), phenylalanine (F), glycine (G), histidine (H), isoleucine (I), lysine (K), leucine (L), methionine (M), asparagine (N), proline (P), glutamine (Q), arginine (R), serine (S), threonine (T), valine (V), tryptophan (W), and tyrosine (Y), as well as non- conventional amino acids such as isomers and modifications of the conventional amino acids, e.g., D-amino acids, non-protein amino acids, post-translationally modified amino acids, enzymatically modified amino acids, ^-amino acids, constructs or structures designed to mimic amino acids (e.g., ^,^-disubstituted amino acids, N-alkyl amino acids, lactic acid, ^-alanine, naphthylalanine, 3-pyridylalanine, 4-hydroxyproline, O-phosphoserine, N-acetylserine, N- formylmethionine, 3-methylhistidine, 5-hydroxylysine, and nor-leucine), and other non- conventional amino acids, as described, for example, in U.S. Pat. No.5,679,782 to Rosenberg et al. The polypeptides described herein may include one or more non-natural amino acids bearing a functional group that enables conjugation to a secondary antigen, e.g., a polysaccharide. Polypeptides can be (a) naturally occurring, (b) produced by chemical synthesis, (c) produced by recombinant DNA technology, (d) produced by biochemical or enzymatic fragmentation of larger molecules, (e) produced by methods resulting from a combination of methods (a) through (d) listed above, or (f) produced by any other means for producing peptides, such as cell-free protein synthesis.

[0019] In some embodiments, the immunogenic compositions provided herein comprise polynucleotides encoding the VZV and RSV polypeptide antigens. In some embodiments, the polynucleotide is an mRNA polynucleotide comprising an open reading frame (ORF) encoding the VZV and / or RSV polypeptide antigens. In some embodiments, the immunogenic composition comprises at least two mRNA polynucleotides, wherein the first mRNA polynucleotide comprises an ORF encoding a VZV polypeptide antigen and the second mRNA polynucleotide comprises an ORF encoding an RSV polypeptide antigen. 4 311398121

[0020] Naturally-occurring eukaryotic mRNA molecules can contain stabilizing elements, including, but not limited to untranslated regions (UTR) at their 5′-end (5′ UTR) and / or at their 3′-end (3′ UTR), in addition to other structural features, such as a 5′-cap structure or a 3′-poly(A) tail. Both the 5′ UTR and the 3′ UTR are typically transcribed from the genomic DNA and are elements of the premature mRNA. Characteristic structural features of mature mRNA, such as the 5′-cap and the 3′-poly(A) tail are usually added to the transcribed (premature) mRNA during mRNA processing. In some embodiments, the aforementioned mRNAs may further comprise a 5′ cap (e.g., 7mG(5′)ppp(5′)NlmpNp), a polyA tail (e.g., ˜100 nucleotides), or a 5′ cap and a polyA tail.

[0021] “Polynucleotides” comprise a polymer of nucleotides (nucleotide monomers). Polynucleotides may be or may include, for example, deoxyribonucleic acids (DNAs), ribonucleic acids (RNAs), threose nucleic acids (TNAs), glycol nucleic acids (GNAs), peptide nucleic acids (PNAs), locked nucleic acids (LNAs, including LNA having a β-D-ribo configuration, α-LNA having an α-L-ribo configuration (a diastereomer of LNA), 2′-amino- LNA having a 2′-amino functionalization, and 2′-amino-α-LNA having a 2′-amino functionalization), ethylene nucleic acids (ENA), cyclohexenyl nucleic acids (CeNA) and / or chimeras and / or combinations thereof.

[0022] Messenger RNA (mRNA) is any ribonucleic acid that encodes a (at least one) protein (a naturally-occurring, non-naturally-occurring, or modified polymer of amino acids) and can be translated to produce the encoded protein in vitro, in vivo, in situ, or ex vivo. The skilled artisan will appreciate that, except where otherwise noted, nucleic acid sequences set forth in the instant application may recite “T”'s in a representative DNA sequence but where the sequence represents RNA (e.g., mRNA), the “T”'s would be substituted for “U”s.

[0023] An open reading frame (ORF) is a continuous stretch of DNA or RNA beginning with a start codon (e.g., methionine (ATG or AUG)) and ending with a stop codon (e.g., TAA, TAG or TGA, or UAA, UAG or UGA). An ORF typically encodes a protein. It will be understood that the polynucleotide sequences referenced herein may further comprise additional elements, e.g., 5′ and 3′ UTRs, but that those elements, unlike the ORF, need not necessarily be present in an immunogenic composition of the present disclosure.

[0024] The terms “sequence identity,” “percent sequence homology,” and “sequence homology,” in the context of a polypeptide sequence, refer to two or more sequences that are the same or have a specified percentage of amino acid residues (or nucleotides) that are the 5 311398121same, when compared and aligned for maximum correspondence over a given length (comparison window), as measured using a sequence comparison algorithm, e.g., BLASTP or the Smith-Waterman homology search algorithm. In the present context, the percent sequence homology may be determined over the full-length of the polypeptide or just a portion. One method for calculating percent sequence homology is the BLASTP program having its defaults set at a wordlength (W) of 3, an expectation (E) of l 0, and the BLOSUM62 scoring matrix; see, e.g., Henikoff et al. (1989) Proc. Natl. Acad. Sci. USA 89:10915. Exemplary determination of sequence alignment and % sequence identity employs the BESTFIT or GAP programs in the GCG Wisconsin Software package (Accelrys, Madison Wis.), using the default parameters provided. If these preferred methods of calculating sequence identity give differing amounts, the method giving the higher sequence identity controls. The term “substantially homologous” refers to a percent sequence homology over a given length (e.g., “x” amino acids of a polypeptide) of at least about 50%, thus including, for example, at least about 75%, at least about 80%, at least about 90%, at least about 95%, at least about 97%, at least about 99%, and 100%.

[0025] The VZV and / or RSV polypeptide antigen can be the full-length protein, or a portion of the protein so long as the portion selected results in a polypeptide fragment that possesses the ability to generate a therapeutic or prophylactic immunogenic response to infection with VZV and / or RSV. Usually, these immunogenic portions or fragments of the full protein are at least 20 amino acid residues in length. Provided the desired immunogenic properties are maintained, the length of the polypeptide antigen is a matter of design choice and can be at least 20, 25, 30, 35, 40, 45, 50, 60, 70, 80, 90, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, or 600 or more amino acid residues, up to and including the full-length protein. The polypeptide antigen may not be an exact copy of the native protein to which it corresponds. Additions, deletions, and substitutions (often conservative substitutions) can also occur provided useful immunogenic properties are retained. Routine testing in animals or humans can demonstrate readily whether an polypeptide antigen synthesized as described herein generates a therapeutic or prophylactic immunogenic response to infection by VZV and / or RSV.

[0026] In some embodiments, the present disclosure provides a purified polypeptide antigen. As used herein, when the term “purified” is used in reference to a molecule, it means that the concentration of the molecule being purified has been increased relative to the concentration of the molecule in its natural environment. The term may also refer to 6 311398121purification of a chemically synthesized molecule from a reaction mixture in which the molecule has been generated as a reaction product. As used herein, when the term “isolated” is used in reference to a molecule, the term means that the molecule has been removed from its native environment. For example, a polynucleotide or a polypeptide naturally present in a living organism is not “isolated,” but the same polynucleotide or polypeptide separated from the coexisting materials in its natural state is “isolated.” An isolated moiety, whether separated from a native environment or from a non-natural environment (e.g., recombinant expression, cell-free expression, chemical synthesis, etc.), is preferably at least about 1% pure, 5% pure, 10% pure, 20% pure, 30% pure, 40% pure, 50% pure, 60% pure, 70% pure, 80% pure, 90% pure, 95% pure, or 99% pure, or they may be 100% pure. As used herein, the term “% pure” indicates the percentage of a composition that is made up of the molecule of interest, by weight.

[0027] In some embodiments, the VZV and RSV polypeptide antigens comprises an amino sequence substantially homologous to a wild type (WT) VZV and RSV antigen sequence. In some embodiments, the VZV and RSV polypeptide antigen comprises an amino acid sequence that is at least 75%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 99%, or 100% identical to the WT VZV and RSV polypeptide antigen sequence. Exemplary WT VZV antigens include the VZV glycoprotein E (gE) (Uniprot Q9J3M8), gB (Uniprot Q4JR05), gH (Uniprot P09257) and gL (Uniprot P09257). Exemplary WT RSV antigens include the RSV F antigen from the B1 strain (Uniprot O36634), RSV F antigen from the A2 strain (Uniprot P03420), RSV F antigens from Uniprot P12568, as well as the RSV G antigen (UniProt P20895). See Martinez et al (2021). PLoS ONE 16(2): e0246770. In some embodiments, the RSV F antigen comprises one or more mutations relative to the WT antigen. See e.g., Che et al., Sci. Transl. Med.15, eade6422 (2023), incorporated by reference. In some embodiments, the RSV F antigen is stabilized in the pre-fusion conformation.

[0028] In some embodiments, the VZV antigen is a truncated form of the gE protein. In some embodiments, the truncated form of the gE protein is selected from those listed in Table 1. In particular embodiments, the immunogenic compositions described herein comprise the gE antigen of SEQ ID NO: 1. Additional gE antigens are described in the art. See e.g., SEQ ID NO: 3 of US 2006 / 121052, and SEQ ID NO: 1 of WO 2006 / 094756. Table 1: Exemplary VZV gE Antigens7 311398121

[0029] In some embodiments, the immunogenic compositions provided herein comprise at least 25 µg of a VZV gE antigen. In some embodiments, the immunogenic compositions provided herein comprise between 25 µg and 125 or higher concentration µg of a VZV gE antigen. In some embodiments, the immunogenic compositions provided herein comprise 25 µg, 30 µg, 35 µg, 40 µg, 45 µg, 50 µg, 55 µg, 60 µg, 65 µg, 70 µg, 75 µg, 80 µg, 85 µg, 90 µg, 95 µg, 100 µg, 105 µg, 110 µg, 115 µg, 120 µg, or 125 µg of a VZV gE antigen. In some embodiments, the immunogenic compositions provided herein comprise about 50 µg of a VZV gE antigen. In some embodiments, the immunogenic compositions provided herein comprise about 45 µg, 46 µg, 47 µg, 48 µg, 49 µg, 50 µg, 51 µg, 52 µg, 53 µg, 54 µg, or 55 µg of a VZV gE antigen. In some embodiments, the immunogenic compositions provided herein comprise about 75 µg of a VZV gE antigen. In some embodiments, the immunogenic compositions provided herein comprise about 70 µg, 71 µg, 72 µg, 73 µg, 74 µg, 75 µg, 76 µg, 77 µg, 78 µg, 79 µg, or 80 µg of a VZV gE antigen. In some embodiments, the immunogenic compositions provided herein comprise about 100 µg of a VZV gE antigen. In some embodiments, the immunogenic compositions provided herein comprise 95 µg, 96 µg, 97 µg, 98 µg, 99 µg, 100 µg, 101 µg, 102 µg, 103 µg, 104 µg, or 105 µg of a VZV gE antigen. In some embodiments, the VZV gE antigen comprises SEQ ID NO: 1. In some embodiments, the VZV gE antigen consists of SEQ ID NO: 1.

[0030] In some embodiments, the RSV antigen is the pre-fusion conformation of the F-antigen (also referred to as PreF3). In some embodiments, the RSV antigen is the RSV G antigen. In some embodiments, the RSV antigen is a variant RSV F antigen comprising one or more mutations relative to the WT RSV F A2 antigen (SEQ ID NO: 3). In some embodiments, the mutations comprise one or more of the mutations from Table 1 of Che et al., Sci. Transl. Med. 15, eade6422 (2023). Exemplary RSV F antigen sequences are provided in Table 2 below. 8 311398121Table 2: Exemplary RSV F Antigens

[0031] In some embodiments, the immunogenic compositions provided herein comprise at least 60 µg of an RSV antigen. In some embodiments, the immunogenic compositions provided herein comprise between 60 µg and 180 µg of an RSV antigen. In some 9 311398121embodiments, the immunogenic compositions provided herein comprise 60 µg, 65 µg, 70 µg, 75 µg, 80 µg, 85 µg, 90 µg, 95 µg, 100 µg, 105 µg, 110 µg, 115 µg, 120 µg, 125 µg, 130 µg, 135 µg, 140 µg, 145 µg, 150 µg, 155 µg, 160 µg, 165 µg, 170 µg, 175 µg, or 180 µg of an RSV antigen.

[0032] In some embodiments, the immunogenic compositions provided herein comprise at least 60 µg of an RSV F-antigen. In some embodiments, the immunogenic compositions provided herein comprise between 60 µg and 180 µg of an RSV F-antigen. In some embodiments, the immunogenic compositions provided herein comprise 60 µg, 65 µg, 70 µg, 75 µg, 80 µg, 85 µg, 90 µg, 95 µg, 100 µg, 105 µg, 110 µg, 115 µg, 120 µg, 125 µg, 130 µg, 135 µg, 140 µg, 145 µg, 150 µg, 155 µg, 160 µg, 165 µg, 170 µg, 175 µg, or 180 µg of an RSV F-antigen. In some embodiments, the RSV F antigen comprises any one of SEQ ID NO: 2-4. In some embodiments, the RSV F antigen consists of any one of SEQ ID NOs: 2-4.

[0033] In some embodiments, the RSV F antigen comprises SEQ ID NO: 2. In some embodiments, the RSV F antigen consists of SEQ ID NO: 2. In some embodiments, the RSV F antigen comprises SEQ ID NO: 3. In some embodiments, the RSV F antigen consists of SEQ ID NO: 3. In some embodiments, the RSV F antigen comprises SEQ ID NO: 4. In some embodiments, the RSV F antigen consists of SEQ ID NO: 4.

[0034] In some embodiments, the immunogenic compositions provided herein comprise the VZV gE antigen of SEQ ID NO: 1 and the RSV F antigen of SEQ ID NO: 2. In some embodiments, the immunogenic compositions provided herein comprise the VZV gE antigen of SEQ ID NO: 1 and the RSV F antigen of SEQ ID NO: 3. In some embodiments, the immunogenic compositions provided herein comprise the VZV gE antigen of SEQ ID NO: 1 and the RSV F antigen of SEQ ID NO: 4.

[0035] In some embodiments, the immunogenic compositions provided herein comprise the VZV gE antigen of SEQ ID NO: 1 and the RSV F antigen of SEQ ID NO: 2 and SEQ ID NO: 3. In some embodiments, the immunogenic compositions provided herein comprise the VZV gE antigen of SEQ ID NO: 1 and the RSV F antigen of SEQ ID NO: 2 and SEQ ID NO: 4. In some embodiments, the immunogenic compositions provided herein comprise the VZV gE antigen of SEQ ID NO: 1 and the RSV F antigen of SEQ ID NO: 3 and SEQ ID NO: 4. 10 311398121Immunogenic Compositions

[0036] In some embodiments, the present disclosure provides immunogenic compositions comprising the RSV and VZV antigens described herein. As used herein, the term “immunogenic” refers to the ability of an antigen (e.g., a polypeptide), to elicit an immune response, either a humoral or cellular immune response, and preferably both. In a preferred embodiment, the subject will display either a therapeutic or protective immunological response to administration of an “effective amount” or “immunologically effective amount” of an immunogenic composition herein such that resistance to new infection will be enhanced and / or the clinical severity of the disease will be reduced. The immunological response will normally be demonstrated by alleviation or elimination of at least one symptom associated with the infection.

[0037] As used herein, the term “vaccine” refers to an immunogenic composition which is used to induce an immune response that provides protective immunity against a pathogen (e.g., immunity that protects a subject against infection with the pathogen and / or reduces the severity of the disease or condition caused by infection with the pathogen). The protective immune response may include formation of antibodies and / or a cell-mediated response.

[0038] The immunogenic compositions may further comprise one or more excipients. The excipients are immunologically and pharmacologically inert components that are “pharmaceutically acceptable.” A “pharmaceutically acceptable” component herein is one that (1) can be included in a immunogenic composition administered to a subject without causing significant unwanted biological effects or interacting in a deleterious manner with any of the other components of the formulation; and (2) meets the criteria set out in the Inactive Ingredient prepared by the U.S. Food and Drug Administration, and, preferably, has also been designated “Generally Regarded as Safe” (“GRAS”). The type of excipient or excipients incorporated into the immunogenic compositions described herein will depend, in part, on the selected mode of administration and the particular formulation type or dosage form, e.g., injectable liquid formulations, intranasal spray formulations, or the like; modes of administration and corresponding formulations are discussed infra. In general, however, inert components that can be advantageously incorporated into the immunogenic compositions described herein include, without limitation, vehicles, solubilizers, emulsifiers, stabilizers, preservatives, isotonicity agents, buffer systems, dispersants, diluents, viscosity modifiers, absorption enhancers, and combinations thereof. A thorough discussion of pharmaceutically acceptable 11 311398121inert additives is available in Gennaro (2000) Remington: The Science and Practice of Pharmacy, 20th Ed., ISBN: 0683306472.

[0039] In some embodiments, the immunogenic compositions described herein are provided as a sterile formulation for administration to a subject, e.g., as a suspension, solution or in lyophilized form to be rehydrated prior to use. Adjuvants

[0040] In some embodiments, the immunogenic composition further comprises one or more adjuvants. As used herein, the term “adjuvant” refers to a compound that, when used in combination with an antigen, augments the immune response to one or more antigens in the immunogenic composition. Augmentation of the immune response may include increasing the antibody titers raised against the one or more antigens, increasing the clonality of the antibody response against the one or more antigens, increasing the intensity of the cellular immune response (e.g.¸ increased memory T cell formation, increased acute cytokine production), and / or diversification of the cellular immune response (e.g., increasing the number of different types of cytokines produced, increasing proliferation of one or more T cell phenotypes, etc.).

[0041] Representative major adjuvant groups suitable for use in the present immunogenic compositions are as follows:

[0042] Mineral salt adjuvants: including alum-based adjuvants such as aluminum phosphate, aluminum hydroxide, aluminum sulfate, aluminum hydrophospho sulfate as well as other mineral salt adjuvants such as the phosphate, hydroxide, and sulfate salts of calcium, iron, and zirconium. Such adjuvants also include Nano Alum (Lu Y, Liu G. Nano alum: A new solution to the new challenge. Hum Vaccin Immunother.2022 Nov 30;18(5));

[0043] Saponin formulations: including the Quillaia saponin Quil A and the Quil A- derived saponin QS-21, Matrix-MTM(Novavax), as well as immune stimulating complexes (ISCOMs) formed upon admixture of cholesterol, phospholipid, and a saponin;

[0044] Bacteria-derived and bacteria-related adjuvants: including, without limitation, cell wall peptidoglycans and lipopolysaccharides derived from Gram negative bacteria such as Mycobacterium spp., Corynebacterium parvum, C. granulosum, Bordetella pertussis, and Neisseria meningitis, such as Lipid A, monophosphoryl Lipid A (MPLA, also referred to as MPL), other Lipid A derivatives and mimetics (e.g., RC529), enterobacterial lipopolysaccharide (“LPS”), TLR4 ligands, and trehalose dimycolate (“TDM”); 12 311398121

[0045] Muramyl peptides: such as N-acetyl muramyl-L-alanyl-D-isoglutamine (“MDP”) and MDP analogs and derivatives, e.g., threonyl-MDP and nor-MDP;

[0046] Oil-based adjuvants: including oil-in-water (O / W) and water-in-oil (W / O) emulsions, such as squalene-water emulsions (e.g., MF59® (also known as MF59C.1), AS03, AF03), complete Freund’s adjuvant (“CFA”) and incomplete Freund’s adjuvant (“IFA”);

[0047] Liposome adjuvants: Microsphere adjuvants formed from biodegradable and non-toxic polymers such as a poly(^-hydroxy acid), a poly(hydroxy butyric) acid, a polyorthoester, a polyanhydride, a polycaprolactone, etc.;

[0048] Nanostructured lipid carriers (NLCs): Hybrid formulation between oil-in-water (o / w) emulsions and solid lipid nanoparticles (SLNs). The nanoparticle core consists of a liquid oil phase, such as squalene, with a solid phase lipid composed of a saturated triglyceride (Erasmus et al., A Nanostructured Lipid Carrier for Delivery of a Replicating Viral RNA Provides Single, Low-Dose Protection against Zika. Mol Ther.2018 Oct 3;26(10):2507-2522).

[0049] Human immunomodulators: including cytokines, such as interleukins (e.g. IL- 1, IL-2, IL-4, IL-5, IL-6, IL-7, IL-12), interferons (e.g. interferon-^), macrophage colony stimulating factor, and tumor necrosis factor;

[0050] Bioadhesives and mucoadhesives: such as chitosan and derivatives thereof and esterified hyaluronic acid and microspheres or mucoadhesives, such as cross-linked derivatives of poly(acrylic acid), polyvinyl alcohol, polyvinyl pyrrolidone, polysaccharides and carboxymethylcellulose;

[0051] Imidazoquinolone compounds: including Imiquamod and homologues thereof, e.g., Resiquimod;

[0052] TLR-9 agonists: such as Hsp90 and oligodeoxynucleotides containing unmethylated CpG motifs (see, e.g., Bode et al. (2011) Expert Rev. Vaccines 10(4): 499-511). Additional TLR-9 agonists include CpG 1018®, which is a 22-mer oligonucleotide sequences containing CpG motifs that are active in both human and rodents, and PF-03512676 (also known as CpG oligonucleotide 7909), which is a 24mer oligonucleotide; and

[0053] Carbohydrate adjuvants: including the inulin-derived adjuvants gamma inulin and algammulin, and other carbohydrate adjuvants such as polysaccharides based on glucose and mannose, including glucans, dextrans, lentinans, glucomannans, galactomannans, levans, and xylans. 13 311398121

[0054] Synthetic derivatives: (a) GLA – a synthetic lipid A derivative and a TLR4 agonist (Coler et al. A synthetic adjuvant to enhance and expand immune responses to influenza vaccine. PLoS One. 2010;5:e13677). Can be provided as a squalene oil-in-water emulsion formulation, formulated in QS-21 containing liposomes (GLA-LSQ), or an aqueous formulation (GLA-AF); (b) SLA - a second-generation lipid adjuvant (SLA) is a synthetic hexa-acylated lipid. See U.S. 10,632,191; U.S. 9,480,740; U.S. 9,814,772; U.S. 8,722,064; and U.S. 10,940,198. Can be provided as a squalene oil-in-water emulsion formulation (SLA-SE), formulated in QS-21 containing liposomes (SLA-LSQ), or an aqueous formulation (SLA-AF); (c) 3M-052 – a synthetic TLR7 / 8 ligand (D. Smirnov et al., Vaccine adjuvant activity of 3M–052: an imidazoquinoline designed for local activity without systemic cytokine induction, Vaccine, 29 (33) (2011), pp. 5434-5442. Can be provided in a in squalene nanoemulsion (3M-052-SE), adsorbed to aluminum (3M-052-Alum), or as an aqueous formulation (3M-052-AF)

[0055] In some embodiments, the immunogenic composition comprises a combination of one or more adjuvants. The combination of the one or more adjuvants can be of the same group (e.g. one or more TLR agonists or one or more carbohydrate antigens) or of different groups (e.g., one or more TLR agonists and one or more carbohydrate adjuvants). In some embodiments, the immunogenic composition comprises one or more adjuvants comprising a saponin (e.g., QS-21) and a bacteria derived adjuvant (e.g., MPL).

[0056] In some embodiments, the adjuvant is an AS01 adjuvant (e.g., AS01E or AS01B), which is a combination of MPL and QS-21 encapsulated in a liposome. The AS01 adjuvants are composed of 3-O-desacyl-4’-monophosphoryl lipid A (MPL) from Salmonella minnesota and QS-21, a saponin purified from plant extract Quillaja Saponaria Molina, combined in a liposomal formulation. The liposomes are composed of dioleoyl phosphatidylcholine (DOPC) and cholesterol in a phosphate-buffered saline solution containing disodium phosphate anhydrous, potassium dihydrogen phosphate, sodium chloride, and water for injection.

[0057] In some embodiments, the liposomes are composed of DOPC and cholesterol in a phosphate-buffered saline solution containing trehalose, sodium chloride, potassium dihydrogen phosphate, polysorbate 80, disodium phosphate anhydrous, and water for injection. 14 311398121

[0058] In some embodiments, the adjuvant comprises a combination of an alum-based adjuvant and a TLR-9 agonist adjuvant. In some embodiments, the combination is an alum based adjuvant and CpG 1018. In some embodiments, the adjuvant combination comprises GLA and 3M-052 formulated as a squalene oil-in-water formulation or in liposomes.

[0059] In some embodiments, the immunogenic composition does not comprise an adjuvant and is formulated into a buffer comprising tromethamine, tromethamine hydrochloride, sucrose, mannitol, polysorbate 80, sodium chloride, and water for injection. Administration and Use

[0060] The terms “treat,” “treatment,” and “treating,” as used herein, refer to an approach for obtaining beneficial or desired results, for example, clinical results. For the purposes of this disclosure, beneficial or desired results may include inhibiting or suppressing the initiation or progression of an infection or a disease; ameliorating, or reducing the development of, symptoms of an infection or disease; or a combination thereof.

[0061] “Prevention,” as used herein, is used interchangeably with “prophylaxis” and can mean complete prevention of an infection or disease, or prevention of the development of symptoms of that infection or disease; a delay in the onset of an infection or disease or its symptoms; or a decrease in the severity of a subsequently developed infection or disease or its symptoms.

[0062] In some embodiments, the present disclosure provides methods for immunizing a subject against VZV and / or RSV comprising administering to the subject an effective amount of the immunogenic compositions described herein. In some embodiments, the present disclosure provides methods for reducing the risk of VZV and RSV infection in a subject comprising prophylactically administering to the subject an effective amount of the immunogenic compositions described herein. In some embodiments, the present disclosure provides methods for inducing a protective immune response against VZV and RSV in a subject comprising administering to the subject an effective amount of the immunogenic compositions described herein.

[0063] In some embodiments, provided herein are the use of the immunogenic compositions described herein for immunizing a subject against VZV and RSV. In some embodiments, provided herein are the use of the immunogenic compositions described herein in the manufacture of a medicament for immunizing a subject against VZV and RSV. In some embodiments, provided herein are the use of the immunogenic compositions described herein 15 311398121for reducing the risk of VZV and RSV infection in a subject. In some embodiments, provided herein are the use of the immunogenic compositions described herein in the manufacture of a medicament for reducing the risk of VZV and RSV infection in a subject. In some embodiments, provided herein are the use of the immunogenic compositions described herein for inducing a protective immune response against VZV and RSV in a subject. In some embodiments, provided herein are the use of the immunogenic compositions described herein in the manufacture of a medicament for inducing a protective immune response against VZV and RSV in a subject.

[0064] As used herein, the term “subject” includes humans and other animals. Typically, the subject is a human. For example, the subject may be an adult, a teenager, a child (2 years to 14 years of age), an infant (birth to 2 year), or a neonate (up to 2 months). In particular aspects, the subject is up to 4 months old, or up to 6 months old. In some embodiments, the adults are 50 years or older. In some embodiments, the adults are 55 years or older. In some embodiments, the adults are seniors about 65 years or older, or about 60 years or older. In some embodiments, the subject is a pregnant woman or a woman intending to become pregnant. In other aspects, subject is not a human; for example, a non-human primate; for example, a baboon, a chimpanzee, a gorilla, or a macaque.

[0065] The method may involve administration of the immunogenic composition therapeutically, e.g., to treat a subject suffering from VZV and / or RSV. The method may also involve administration of the immunogenic composition prophylactically, meaning that, for example, the method reduces the risk of VZV and / or RSV infection developing in a subject. When the immunogenic composition is used prophylactically, the subject may be predisposed to a VZV and / or RSV infection as a result of any number of risk factors, including age and / or whether or not the subject is immunocompromised.

[0066] The “immunologically effective amount” or “effective amount” of the immunogenic composition is an amount that, either as a single dose or as part of a series of two or more doses, is effective for treating or preventing VZV and / or RSV infection. The amount administered will vary according to several factors, including the overall health and physical condition of the subject, the subject’s age, the capacity of the subject’s immune system to synthesize relevant antibodies, the form of the composition (e.g., injectable liquid, nasal spray, etc.), and other factors known to the medical practitioner overseeing administration. 16 311398121

[0067] Herein, the term “protective immune response” encompasses eliciting an anti- VZV and / or RSV antibody response in the subject. Antibody titers generated after administration of the immunogenic compositions described herein can be determined by means known in the art, for example by ELISA assays of serum samples derived from immunized subjects.

[0068] Administration of the immunogenic composition can be carried out using any effective mode of systemic delivery. The composition is usually administered parenterally, such as by injection, including intravenous, intramuscular, intraperitoneal, interstitial, or subcutaneous injection; injection may also be gingival, in which case the immunogenic composition is injected directly into the gum. The composition may, in addition, be administered transmucosally, such as via the intranasal, sublingual, transbuccal, intravaginal, or intrarectal routes. Other modes of administration are also envisioned, however, and the invention is not limited in this regard. By way of example, other modes of administration include oral and transdermal delivery as well as administration via inhalation or using a subdermal implant.

[0069] The mode of administration largely dictates the type of formulation or dosage form that comprises the immunogenic composition. Compositions formulated for parenteral administration include sterile aqueous and nonaqueous solutions, suspensions, and emulsions. Injectable aqueous solutions contain the active agent in water-soluble form. Examples of nonaqueous solvents or vehicles include fatty oils, such as olive oil and corn oil, synthetic fatty acid esters, such as ethyl oleate or triglycerides, low molecular weight alcohols such as propylene glycol, synthetic hydrophilic polymers such as polyethylene glycol, liposomes, and the like. Parenteral formulations may also contain excipients such as solubilizers, emulsifiers, stabilizers, preservatives, isotonicity agents, buffer systems, dispersants, diluents, viscosity modifiers, absorption enhancers, and combinations thereof. Injectable formulations are rendered sterile by incorporation of a sterilizing agent, filtration through a bacteria-retaining filter, irradiation, or heat. They can also be manufactured using a sterile injectable medium. The immunogenic composition or individual components thereof may also be in dried, e.g., lyophilized, form that may be rehydrated with a suitable vehicle immediately prior to administration via injection.

[0070] Of the transmucosal routes, intranasal administration is generally although not necessarily preferred. Intranasal formulations, including intranasally administered immunogenic compositions, are known in the art, and should be formulated with reference to 17 311398121the FDA’s Guidance for Industry: Nasal Spray and Inhalation Solution, Suspension, and Spray Drug Products. Intranasal formulations are liquids, i.e., solutions, emulsions, suspensions, or the like, for administration as sprays, intranasal injections, or drops, and can contain adjuvants and pharmaceutically acceptable excipients as above. Because of the relatively large size of the antigens in the formulation, systemic delivery via the intranasal route requires incorporation of a transmucosal absorption enhancer in the immunogenic composition. Examples of suitable transmucosal absorption enhancers include, without limitation, alkylsaccharides, cyclodextrins, and chitosans; see Maggio (2014) J. Excip. Food Chem. 5(2): 100-12; and Merkus et al. (1999) Adv. Drug Deliv. Rev.36: 41-57. The concentration of enhancer is selected to ensure that an immunologically effective amount of the formulation passes through the nasal membrane and into the systemic circulation at an efficient transport rate. Various anatomical and physiological considerations dictating the composition and nature of an intranasal immunogenic composition are discussed, for example, by Aurora (October 2002) Drug Development & Delivery 2(7), incorporated by reference herein.

[0071] Other modes of administration and corresponding formulations include, without limitation: sublingual administration with a rapidly dissolving dosage form such as a rapidly dissolving tablet; transbuccal administration using a buccal patch or other buccal delivery system; intravaginal administration using a pessary, ointment, or cream; intrarectal delivery using a rectal suppository, ointment, or cream; transdermal administration using a transdermal patch or formulation; subdermal administration with an injected implant or pellet; inhalation using a dry powder pulmonary formulation; and oral administration using an oral dosage form such as a tablet, capsule, or the like.

[0072] As alluded to earlier herein, the immunogenic composition is administered to a subject within the context of an appropriate dosage regimen. The composition may be administered once, or two or more times spaced out over an extended time period. For example, an initial, “prime” dose may be followed by at least one “boost” dose. The time interval between the prime and the subsequent boost dose, and between boost doses, is usually in the range of about 2 to about 24 weeks, more typically in the range of about 2 to 12 weeks, such as 2 to 8 weeks, 3-6 weeks, etc.

[0073] In some embodiments, the immunogenic composition is administered as a single dose, wherein the subject further receives treatment with a second immunogenic composition comprising a VZV antigen, wherein the second immunogenic composition does not comprise an RSV antigen. 18 311398121

[0074] Regardless of the mode of administration, e.g., intramuscular injection, gingival injection, intranasal administration, or the like, the volume of a single dose of the vaccine will generally be in the range of about 1 µL to about 500 µL, typically in the range of about 1 µL to about 250 µL, more typically in the range of about 2.5 µL to about 200 µL, and preferably in the range of about 5 µL to about 150 µL. It will be appreciated that the concentration of total antigen in the immunogenic composition corresponds to an immunologically effective dose of the composition per unit volume, working from the aforementioned dose volume guidelines.

[0075] For ease of use, the immunogenic composition of the invention can be incorporated into a packaged product, or “kit,” including instructions for self-administration or administration by a medical practitioner. The kit includes a sealed container housing a dose of the immunogenic composition, typically a “unit dose” appropriate for a single dosage event that is immunologically effective. The vaccine may be in liquid form and thus ready to administer as an injection or the like, or it may be in another form that requires the user to perform a preparation process prior to administration, e.g., hydration of a lyophilized formulation, activation of an inert component, or the like. The kit may also include two or more sealed containers with the prime dose in a first container and a boost dose in one or more additional containers.

[0076] In some embodiments, the immunogenic composition of the invention is formulated to include an effective dose of both the RSV and VZV antigens to produce an antigen specific immune response in a subject. In some embodiments, the vaccine is formulated as a dose comprising 5 µg to 200 µg of the VZV antigen and 5 µg to 200 µg of the RSV antigen. In some embodiments, the dose comprises about 5 µg, about 10 µg, about 15 µg, about 20 µg, about 25 µg, about 30 µg, about 35 µg, about 40 µg, about 45 µg, about 50 µg, about 55 µg, about 60 µg, about 65 µg, about 70 µg, about 75 µg, about 80 µg, about 85 µg, about 90 µg, about 95 µg, about 100 µg, about 110 µg, about 120 µg, about 130 µg, about 140 µg, about 150 µg, about 160 µg, about 170 µg, about 180 µg, about 190 µg, or about 200 µg of the VZV antigen, and about 5 µg, about 10µg, about 15 µg, about 20 µg, about 25 µg, about 30 µg, about 35 µg, about 40 µg, about 45 µg, about 50 µg, about 55 µg, about 60 µg, about 65 µg, about 70 µg, about 75 µg, about 80 µg, about 85 µg, about 90 µg, about 95 µg, about 100 µg, about 110 µg, about 120 µg, about 130 µg, about 140 µg, about 150 µg, about 160 µg, about 170 µg, about 180 µg, about 190 µg, or about 200 µg of the RSV antigen. 19 311398121

[0077] In some embodiments, the dose comprises 5 µg, 10 µg, 15 µg, 20 µg, 25 µg, 30 µg, 35 µg, 40 µg, 45 µg, 50 µg, 55 µg, 60 µg, 65 µg, 70 µg, 75 µg, 80 µg, 85 µg, 90 µg, 95 µg, 100 µg, 110 µg, 120 µg, 130 µg, 140 µg, 150 µg, 160 µg, 170 µg, 180 µg, 190 µg, or 200 µg of the VZV antigen, and 5 µg, 10µg, 15 µg, 20 µg, 25 µg, 30 µg, 35 µg, 40 µg, 45 µg, 50 µg, 55 µg, 60 µg, 65 µg, 70 µg, 75 µg, 80 µg, 85 µg, 90 µg, 95 µg, 100 µg, 110 µg, 120 µg, 130 µg, 140 µg, 150 µg, 160 µg, 170 µg, 180 µg, 190 µg, or 200 µg of the RSV antigen.

[0078] In some embodiments, the dose comprises 10-450 µg total antigen protein. In some embodiments, the dose comprises about 10 µg, about 20 µg, about 30 µg, about 40 µg, about 50 µg, about 60 µg, about 70 µg, about 80 µg, about 90 µg, about 100 µg, about 110 µg, about 120 µg, about 130 µg, about 140 µg, about 150 µg, about 160 µg, about 170 µg, about 180 µg, about 190 µg, about 200 µg, about 210 µg, about 220 µg, about 230 µg, about 240 µg, about 250 µg, about 260 µg, about 270 µg, about 280 µg, about 290 µg, about 300 µg, about 310 µg, about 320 µg, about 330 µg, about 340 µg, about 350 µg, about 360 µg, about 370 µg, about 380 µg, about 390 µg, about 400 µg, about 410 µg, about 420 µg, about 430 µg, about 440 µg, or about 450 µg total antigen protein. In some embodiments, the dose comprises 10 µg, 20 µg, 30 µg, 40 µg, 50 µg, 60 µg, 70 µg, 80 µg, 90 µg, 100 µg, 110 µg, 120 µg, 130 µg, 140 µg, 150 µg, 160 µg, 170 µg, 180 µg, 190 µg, 200 µg, 210 µg, 220 µg, 230 µg, 240 µg, 250 µg, 260 µg, 270 µg, 280 µg, 290 µg, 300 µg, 310 µg, 320 µg, 330 µg, 340 µg, 350 µg, 360 µg, 370 µg, 380 µg, 390 µg, 400 µg, 410 µg, 420 µg, 430 µg, 440 µg, or 450 µg total antigen protein.

[0079] In some embodiments, the effective amount of the immunogenic composition disclosed is delivered in multiple doses, with each dose comprising 25-225 μg total antigen protein. In some embodiments, the effective amount of the immunogenic composition is split into two or more doses. In some embodiments, each of the multiple doses comprise the same amount of antigen protein. In some embodiments, each of the multiple doses comprise differing amounts of the antigens, but together the doses comprise 10-450 µg antigen protein

[0080] Exemplary combinations of the two antigens are shown in Table 2 below. In some embodiments, the combined amounts of the VZV gE antigen (SEQ ID NO: 1) and the RSV antigen (SEQ ID NOs: 2-4) are selected from any one of combinations 1-400 in Table 2. 20 311398121og µ d09 992949698 9n09129149169189109229249 9 9 9 9 9 9262820323436383ane 8 8 8 8 8 8 8 8 8 8 8 8 8 8g0i882848688t0 2 4 6 88 81 1 1 1 102224262820323436383n a07 772747678 7072747 7 7 7 7 7 7 7 7 7 7 7VS1 1 1618102224262820323436383Rfo06 662646668 6g06 6 6 6 6 6 6 6 6 6 6 6 6 612141618102224262820323436383µfo05 552545655 5 5 5 5 5 5 5 5 5 5 5 5 5 5s 80n12141618102224262820323436383oita04 4 4n442444648i0124144164184104224244264284204324344364383b mo033 3 33 3 3 3 3 3 3 3 3 3 3 3 3 3 3c3 2 4 638012141618102224262820323436383yral02 222242628 202224262820222426282 2 2 2 2p1 1 1 1 1 2 2 2 2 20323436383mex EV01 112141618 1:S 01 1 1 1 1 1 1 1 1 1 1 1 1 1121416181022242628203234363832RelbV0aZ10203040506070809 000 0 0 0 0 0 0 0 0 0111213141516171819102TV311398121 21

[0081] In some embodiments, the immunogenic composition comprises two or more antigens in a single formulation. In some embodiments, the amount of the two or more antigens in the immunogenic composition comprises a lower dose of the two or more antigens than an immunogenic composition that comprises any one of the two or more antigens. In some embodiments, the immunogenic composition comprising two or more antigens comprises less adjuvant than an immunogenic composition that comprises any one of the two or more antigens. In some embodiments, the immunogenic composition comprises 20 µg SLA-SE. In some embodiments, the immunogenic composition comprises 15 µg SLA-SE. In some embodiments, the immunogenic composition comprises 10 µg SLA-SE. In some embodiments, the immunogenic composition comprises 5 µg SLA-SE. In some embodiments, the immunogenic composition comprising two or more antigens does not comprise an adjuvant.

[0082] In some embodiments, the immunogenic response of the immunogenic composition that comprises two or more antigens is higher than the immunogenic composition that comprises any one of the two or more antigens. In some embodiments, the immunogenic response of the immunogenic composition that comprises two or more antigens increases the antibody titer as compared to the immunogenic response of an immunogenic composition that comprises any one of the two or more antigens. In some embodiments, the immunogenic response of the immunogenic composition that comprises two or more antigens induces a more polyclonal antibody response as compared to the immunogenic response of an immunogenic composition that comprises any one of the two or more antigens. In some embodiments, the immunogenic response of the immunogenic composition that comprises two or more antigens increases production of memory T cells as compared to the immunogenic response of an immunogenic composition that comprises any one of the two or more antigens. In some embodiments, the immunogenic response of the immunogenic composition that comprises two or more antigens induces an increased cytokine expression as compared to the immunogenic response of an immunogenic composition that comprises any one of the two or more antigens.

[0083] In some embodiments, the immunogenic composition that comprises two or more antigens induces fewer adverse events as compared to the immunogenic composition that comprises any one of the two or more antigens. In some embodiments, the immunogenic composition that comprises two or more antigens induces fewer grade 3 adverse events as compared to the immunogenic composition that comprises any one of the two or more antigens. In some embodiments, the immunogenic composition that comprises two or more antigens 22 311398121induces fewer grade 2 adverse events as compared to the immunogenic composition that comprises any one of the two or more antigens.

[0084] It is to be understood that while the invention has been described in conjunction with a number of specific embodiments, the foregoing description as well as the experimental section that follows are intended to illustrate and not limit the scope of the invention. In this regard, no attempt is made to show details of the invention in more detail than is necessary for the fundamental understanding of the invention, the description taken with the drawings and / or examples making apparent to those skilled in the art how the invention may be embodied in practice. This disclosure includes all modifications and equivalents of the subject matter recited in the claims appended hereto as permitted by applicable law. Moreover, any combination of the elements of the invention described herein are encompassed by the disclosure unless otherwise indicated herein or clearly contradicted by context. INCORPORATION BY REFERENCE

[0085] All references, articles, publications, patents, patent publications, and patent applications cited herein are incorporated by reference in their entireties for all purposes. However, mention of any reference, article, publication, patent, patent publication, and patent application cited herein is not, and should not be taken as, an acknowledgment or any form of suggestion that they constitute valid prior art or form part of the common general knowledge in any country in the world. Examples Example 1: Immunogenicity study

[0086] Experiments will be performed to assess the immunogenicity of immunogenic compositions comprising a VZV gE antigen (e.g. SEQ ID NO: 1) and an RSV antigen (e.g., SEQ ID NOs: 2, 3, or 4) and an adjuvant (e.g., SLA-SE). Mice are immunized intramuscularly or intradermally with the immunogenic compositions. Sera are collected every three weeks until weeks 33-51. At the experiment endpoint, mice are sacrificed and spleens and lymph nodes harvested. Serum antibody titers against VZV gE and RSV F are determined by ELISA. Sera collected from each mouse during weeks 10-16 are pooled, and total Ig purified. Purified antibodies are used for in vitro protection assays. Antibody responses are measured by ELISA for VZV gE-specific and RSV F-specific antibodies. 23 311398121

[0087] The splenocytes are isolated from each spleen and stimulated with virus-free media, Concanavalin A, or VZV or RSV peptides. Secreted cytokines (e.g., IFNγ, IL-2, IL-12, CD40L and TNFα) are measured by fluorospot, ELISPOT and intracellular cytokine staining combined with flow cytometry analysis.

[0088] Sera from the immunized mice are serially diluted in a 96-well plate in triplicate for each condition. RSV and VZV are separately added to wells and incubated for 1 hour to allow for neutralization of the virus inoculum. The neutralized virus / sera mix is then added to Madin-Darby Canine Kidney (MDCK). After 1 hour the infection procedure is stopped and additional sera / antibody mix are added to the cells, and cells are grown for 48-72 hours to allow time for several rounds of the full viral life cycle in the presence of antibodies. Finally, the supernatant is used in a hemagglutination assay with red blood cells to determine the presence of virus and the degree of neutralization.. Example 2: Varicella and Herpes Zoster Animal Models

[0089] Experiments are performed to test the efficacy of the immunogenic compositions described in Example 1 in protection from VZV infection.

[0090] Mice are immunized intramuscularly (IM) or intradermally (ID) at week 0 and week 3 with the immunogenic composition from Example 1. Control groups are immunized with antigen-alone, adjuvant-alone, or vehicle control. At week 7, the animals are challenged with live-attenuated VZV (1 x 105– 2 x 106PFU). At days 3, 7, and 14 post-challenge sera is taken from the mice. At the study endpoint, sera is collected from each mouse. Sera are analyzed for gE antibody responses.

[0091] Additional animal models of varicella and herpes zoster are described in Haberthur et al Pathogens.2013 May 13;2(2):364–382, incorporated herein by reference. For example, guinea pigs can be used to evaluate the effectiveness of the immunogenic compositions described herein. Immunogenic compositions are administered to Hartley guinea pigs at week 0 and week 3. Live attenuated Varicella zoster virus vaccine (15000 PFU / 0.5 mL) is prepared and subcutaneously or intramuscularly administered 1-2 weeks after final virus challenge. Sera are collected throughout the study for gE antibody responses and viral load is determined throughout the study. 24 311398121Example 3: RSV Rodent Model

[0092] Experiments are performed to test the efficacy of the immunogenic compositions described in Example 1 in protection from RSV infection. Animal models of RSV are summarized in Taylor, Vaccine, 35 (2017) 469-480, incorporated herein by reference.

[0093] Briefly, mice (e.g., Balb / c) or cotton rats are immunized intramuscularly with the immunogenic compositions described in Example 1 on days 0 and 28. Mice are challenged intranasally with RSV strain A2(1 x 105– 2 x 106PFU, See Che et al., Sci. Transl. Med.15, eade6422 (2023)). At days 3, 7, and 14 post-infection sera is taken from the mice and viral load is determined via quantitative PCR. Anti-RSV F or other antigen titers are measured via ELISA analysis. Neuralization antibody titers (NT50) are quantified. Animals displaying severe illness as determined by >30% weight loss, extreme lethargy or paralysis are euthanized. Body temperature and weight are assessed and recorded daily. At the study endpoint, viral load is determined for, and sera is collected from, each mouse. Lung tissue is harvested for pathology. Spleens and lymph nodes are harvested for T cell analysis and cytokine production by ELISA / ELISPOT / intracellular cytokine staining. 25 311398121

Claims

Claims 1. An immunogenic composition comprising: (a) a Varicella-zoster virus (VZV) gE glycoprotein comprising the amino acid sequence of SEQ ID NO: 1 or a polynucleotide encoding the same; (b) a Respiratory Syncytial Virus (RSV) antigen or a polynucleotide encoding the same; and (c) an adjuvant.

2. The immunogenic composition of claim 1, wherein the RSV antigen comprises a pre- fusion form of one or more RSV glycoproteins.

3. The immunogenic composition of claim 2, wherein the one or more RSV antigens comprise the F glycoprotein.

4. The immunogenic composition of claim 3, wherein the RSV glycoprotein F comprises the amino acid sequence of any one of SEQ ID NOs: 2-4.

5. The immunogenic composition of any one of claims 1-4, comprising between 25 µg and 125 µg of the VZV gE antigen.

6. The immunogenic composition of any one of claims 1-5, comprising about 100 µg of the VZV gE antigen.

7. The immunogenic composition of any one of claims 1-6, comprising between 60 µg and 180 µg of the RSV antigen.

8. The immunogenic composition of any one of claims 1-7, comprising about 120 µg of the RSV antigen.

9. The immunogenic composition of claim 8, wherein the RSV antigen is the prefusion form of glycoprotein F.

10. The immunogenic composition of any one of claims 1-7, comprising about 60 µg each of two RSV antigens.

11. The immunogenic composition of claim 10, wherein the two RSV antigens are the prefusion forms of glycoprotein F from the A2 and B1 strains (SEQ ID NOs: 3 and 4). 26 31139812112. The immunogenic composition of any one of claims 1-11, wherein the adjuvant is selected from aluminum hydroxide, aluminum phosphate, aluminum hydrophosphate sulfate, MPL, QS-21, MF59C.1, CpG 1018, Matrix-M, MF59, PF-03512676, Matrix-M-2, Vaxfectin, GLA-SE, SLA-SE, 3M-052-SE, GLA-LSQ, SLA-LSQ, 3M-052-Alum, GLA-AF, SLA-AF, 3M-052-AF, GLA-3M-052-LS, NanoAlum, a nanostructured lipid carrier (NLC), or a combination thereof.

13. The immunogenic composition of any one of claims 1-11, wherein the adjuvant is SLA- SE.

14. A method of stimulating an immune response against Varicella Zoster Virus (VZV) and / or Respiratory Syncytial Virus (RSV) in a subject comprising administering the immunogenic composition of any one of claims 1-13.

15. A method of preventing infection with Varicella Zoster Virus (VZV) and / or Respiratory Syncytial Virus (RSV) in a subject comprising administering the immunogenic composition of any one of claims 1-13.

16. The method of claim 14 or 15, wherein a single dose of the immunogenic composition is administered.

17. The method of claim 16, further comprising administering a second immunogenic composition comprising the VZV gE antigen and an adjuvant.

18. The method of claim 17, wherein the second immunogenic composition is administered 2-6 months after administration of the first immunogenic composition.

19. The method of claim 14 or 15, wherein two or more doses of the immunogenic composition are administered.

20. The method of claim 19, wherein the second dose is administered 2-6 months after the first dose. 27 311398121