Peptides comprising stapd or KEKR and methods of use thereof

Abstract

Disclosed are peptides comprising a portion of LukH, wherein the portion of LukH comprises the amino acid sequence of STAPD (SEQ ID NO: 1), KEKR, or both. Disclosed are compositions comprising a peptide comprising, consisting essentially of, at least 90% identical to the amino acid sequence TNKDKNSTAPDDIGKNGK, KDKNSTAPDDIGKNGKI, STAPDDIGKNGKITKRT, EHVDKSQQKEKRNVTNK, KSQQKEKRNTNKDKNS, or a peptide equivalent thereof, wherein the immunogenic equivalent thereof is a peptide comprising, consisting essentially of, or consisting of an amino acid sequence at least 60, 65, 70, 75, 80, 85, 90, 95, or 99% identical to the amino acid sequence EHVDKSQQKEKRNVTNK or KSQQKEKRNTNKDKNS. Disclosed are methods of using the disclosed peptides and compositions, such as, treating or preventing Staphylococcus aureus by administering one or more of the disclosed peptides, VLPs displaying the peptide sequences, nucleic acid sequences, vectors, or compositions to a subject in need thereof.

Description

ATTORNEY DOCKET NO. 37759.0626P1PEPTIDES COMPRISING STAPD OR KEKR AND METHODS OF USE THEREOFCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. Provisional Patent Application No. 63 / 680.389, filed on August 7, 2024, which is incorporated by reference herein in its entirety.STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH

[0002] This invention was made with government support under VA Merit award I01BX004720 awarded by the Department of Veterans Affairs. The government has certain rights in the invention.REFERENCE TO SEQUENCE LISTING

[0003] The Sequence Listing submitted August 7. 2025 as a text file named “37759.0626Pl.xml,” created on August 1, 2025, and having a size of 16,431 bytes is hereby incorporated by reference pursuant to 37 C.F.R. § 1.52(e)(5).BACKGROUND

[0004] The discovery and delineation of protective neutralizing epitopes can inform the rational design and development of vaccines and immunotherapeutics that can precisely focus immune responses towards critical pathogen sequences. This epitope-focused vaccine approach may be especially valuable for developing vaccines for pathogens where conventional approaches have failed to elicit protective humoral responses including HIV, malaria and RSV. Such an approach may also be enabling for developing vaccines against pathogens, like Staphylococcus aureus, where the elicitation of humoral immunity against multiple critical epitopes across an array of virulence factors may be required for protective efficacy.

[0005] 5. aureus causes skin and soft tissue infections, pneumonia, bacteremia and toxin- mediated syndromes with substantial morbidity and mortality. Antibiotics are critical to the management of such infections, but resistance is limiting efficacy. Though methicillin-resistant strains of S. aureus first appeared in healthcare settings (HA-MRSA), the emergence of community-associated MRSA (CA-MRSA), which frequently infects otherwise healthy individuals, has ushered in a new, more dangerous phase in the evolution of this pathogen. Pulse field gel electrophoresis type USA300, is the most common cause of skin and soft tissue infections in the community and is an important cause of sepsis and necrotizing pneumonia.

[0006] S. aureus employs a diverse repertoire of surface-associated virulence factors including protein A, clumping factor A. IsdB and capsular polysaccharides, and secreted virulence factors including alpha hemolysin, also known as alpha toxin (AT), beta, gamma andATTORNEY DOCKET NO. 37759.0626P1 delta hemolysins, the phenol soluble modulins, and the leukotoxins. The presence of this multitude of virulence factors suggests that a successful vaccine for S. aureus may require the effective targeting of more than one virulence factor, perhaps employing a poly-virulence factor, multivalent vaccine.

[0007] Among the secreted virulence factors, alpha hemolysin or alpha toxin (AT) has long been considered a critical virulence factor in human infections. AT is a 293 residue protein that binds ADAM 10 at the cell surface, and then self-associates to form a heptameric structure that ultimately creates a pore in eukaryotic membranes. AT has wide-ranging effects on a variety of human cell types including monocytes, B and T cells and epithelial cells and appears to play a critical role in subverting both the innate and adaptive immune responses to S. aureus infection. The importance of AT in human infections has been strongly bolstered by analyses showing that increased expression of AT plays a prominent role in the increased virulence associated with the US A300 lineage, the most common cause of CA-MRSA infections including SSTIs and invasive systemic disease like sepsis, and may also be critical to the exceptional virulence associated with USA100 strains which are a serious cause of invasive disease and which represent the majority of VISA isolates. The primacy of AT has been demonstrated experimentally in the mouse models of S. aureus pneumonia, peritonitis and dermonecrosis where, in contrast to WT strains, gene-deletion mutant strains incapable of producing functional AT are virtually devoid of virulence.

[0008] Unlike the case with AT, mice are relatively resistant to the effects of leukotoxins, a fact that delayed development of a clear understanding of the importance of these toxins in S. aureus-mediated disease. The rabbit model, however, as well as in vitro studies with rabbit and human leukocytes, has now illuminated the critical contributions of the leukotoxins including LukED, PVL, gamma toxin and LukGH, to S. aureus infections in rabbits and humans, where the predominant effects are to create a highly redundant system for disrupting innate immunity by impairing and killing leukocytes. Among the leukotoxins, LukGH appears to be particularly important, with one study utilizing human neutrophils demonstrating that PVL and LukGH synergize to account for all the LAC-mediated neutrophil cytotoxicity in vitro. Recent work has revealed the critical importance of LukGH, together with AT, in the development of MRS A- mediated secondary pneumonias post influenza infection. Studies in children with invasive S. aureus disease have revealed significant levels of LukGH-neutralizing Ab in convalescent serum, suggesting that LukGH may be an important target for adaptive immunity’, and potentially for vaccine development.

[0009] In the anthrax and S. aureus models, one can map linear toxin neutralizing epitopesATTORNEY DOCKET NO. 37759.0626P1 in vivo using peptide immunogens, in concert with available crystal structure and an optimized in vitro toxin neutralization assay. The highly solvent-exposed residues within the functionally important random coil structure at the amino terminus of AT led us to hypothesize that this segment might contain a linear B cell determinant with potential as a protective neutralizing epitope, and prior studies confirmed the presence of such an epitope referred to as the N- terminal determinant (NTD). Considering the homology in both the monomeric and assembled toxin forms for AT and the leukotoxins, which are all pore-forming toxins (PFTs), it is thought that LukGH, like AT, can also have a neutralizing epitope in the N-terminal region of the full- length polypeptides, LukG and LukH, which together constitute the LukGH heterodimer.BRIEF SUMMARY

[0010] Disclosed are peptides, wherein the peptides comprise 5 to 50 amino acids, wherein the amino acid sequence comprises STAPD (SEQ ID NO: 1).

[0011] Disclosed are peptides, wherein the peptides comprise 6 to 50 amino acids, wherein the amino acid sequence comprises STAPDD (SEQ ID NO: 2).

[0012] Disclosed are peptides comprising, consisting essentially of, or consisting of a STAPD (SEQ ID NO: 1) epitope such as, but not limited to, STAPDD (SEQ ID NO:2). TNKDKNSTAPDDIGKNGK (SEQ ID NO:3), KDKNSTAPDDIGKNGKI (SEQ ID NO:4). STAPDDIGKNGKITKRT (SEQ ID NO:5) or an immunogenic equivalent thereof, wherein the immunogenic equivalent thereof is a peptide comprising, consisting essentially of, or consisting of an amino acid sequence at least 60, 65, 70, 75, 80, 85, 90, 95, or 99% identical to STAPDD (SEQ ID NO:2), TNKDKNSTAPDDIGKNGK (SEQ ID NO:3), KDKNSTAPDDIGKNGKI (SEQ ID NO:4), or STAPDDIGKNGKITKRT (SEQ ID NO:5).

[0013] Disclosed are peptides, wherein the peptides comprise 4 to 30 amino acids, wherein the amino acid sequence comprises KEKR (SEQ ID NO:6) or KDKR (SEQ ID NO:7).

[0014] Disclosed are peptides comprising, consisting essentially of, or consisting of a KEKR (SEQ ID NO:6) epitope such as. but not limited to, EHVDKSQQKEKRNVTNK (SEQ ID NO:8), KSQQKEKRNTNKDKNS (SEQ ID NO:9), or an immunogenic equivalent thereof, wherein the immunogenic equivalent thereof is a peptide comprising, consisting essentially of, or consisting of an amino acid sequence at least 60, 65, 70, 75, 80, 85, 90, 95, or 99% identical to EHVDKSQQKEKRNVTNK (SEQ ID NO:8) or KSQQKEKRNTNKDKNS (SEQ ID NO:9).

[0015] Disclosed are modified viral capsid proteins comprising a portion of LukH, wherein the portion of LukH comprises STAPD (SEQ ID NO:1), KEKR (SEQ ID NO:6), or both.

[0016] Disclosed are virus-like particles (VLPs) comprising any one of the modifiedATTORNEY DOCKET NO. 37759.0626P1 proteins disclosed herein used to produce VLPs.

[0017] Disclosed are nucleic acid sequences or constructs comprising a nucleic acid sequence encoding STAPD (SEQ ID NO: 1), STAPDD (SEQ ID NO: 2), TNKDKNSTAPDDIGKNGK (SEQ ID NO:3), KDKNSTAPDDIGKNGKI (SEQ ID NO:4), STAPDDIGKNGKITKRT (SEQ ID NO:5). KEKRNVTNKDKN STAPD (SEQ ID NOTO), KEKR (SEQ ID NO:6), KDKR (SEQ ID NOV), EHVDKSQQKEKRNVTNK (SEQ ID NO: 8), or KSQQI<EKRNTNI<DI<NS (SEQ ID NO:9), or a functional variant thereof.

[0018] Disclosed are compositions comprising a peptide comprising / consisting essentially of the STAPD (SEQ ID NO: 1) epitope STAPDD (SEQ ID NOV). TNKDKNSTAPDDIGKNGK (SEQ ID NOV), KDKNSTAPDDIGKNGKI (SEQ ID NO:4), STAPDDIGKNGKITKRT (SEQ ID NO:5) or an immunogenic equivalent thereof, wherein the immunogenic equivalent thereof is a peptide comprising / consisting essentially of an amino acid sequence at least 90% identical to STAPDD (SEQ ID NOV). TNKDKNSTAPDDIGKNGK (SEQ ID NO: 3), KDKNSTAPDDIGKNGKI (SEQ ID NO:4), or STAPDDIGKNGKITKRT (SEQ ID NO:5).

[0019] Disclosed are compositions comprising a peptide comprising / consisting essentially of the KEKR (SEQ ID NO: 6) epitope such as, but not limited to, EHVDKSQQKEKRNVTNK (SEQ ID NO:8), KSQQKEKRNTNKDKNS (SEQ ID NOV), or an immunogenic equivalent thereof, wherein the immunogenic equivalent thereof is a peptide comprising, consisting essentially of, or consisting of an amino acid sequence at least 60, 65, 70, 75, 80, 85, 90, 95, or 99% identical to EHVDKSQQKEKRNVTNK (SEQ ID NO: 8) or KSQQKEKRNTNKDKNS (SEQ ID NOV).

[0020] Disclosed are methods of neutralizing the cytotoxicity of leukotoxin LukGH in a subject infected with Staphylococcus aureus comprising administering to the subject one or more of the disclosed immunogenic compositions, peptides, nucleic acids, or VLPs, wherein the subject generates neutralizing antibodies (nAbs) against LukH.

[0021] Disclosed are methods of treating a subject infected with Staphylococcus aureus comprising administering to the subject one or more of the disclosed immunogenic compositions, peptides, nucleic acids, or VLPs.

[0022] Disclosed are methods of inducing an immune response against Staphylococcus aureus in a subject, comprising administering to the subject one or more of the disclosed immunogenic compositions, peptides, nucleic acids, or VLPs, wherein the immune response targets Staphylococcus aureus leukotoxin LukH.

[0023] Additional advantages of the disclosed method and compositions will be set forth in part in the description which follows, and in part will be understood from the description, orATTORNEY DOCKET NO. 37759.0626P1 may be learned by practice of the disclosed method and compositions. The advantages of the disclosed method and compositions will be realized and attained by means of the elements and combinations particularly pointed out in the appended claims. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the invention as claimed.BRIEF DESCRIPTION OF THE DRAWINGS

[0024] The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate several embodiments of the disclosed method and compositions and together with the description, serve to explain the principles of the disclosed method and compositions.

[0025] FIG. 1 shows structural models of monomeric alpha toxin (UniProt Q2G1X0), LukG (UniProt A0A7K3N6K1) and LukH (UniProt A0A6G4N894 ).

[0026] FIG. 2 shows structural models of assembled alpha toxin (PDB 3ANZ) and LukGH (PDB 4TW1).

[0027] FIG. 3 shows an alignment of N-terminal segments of S. aureus pore-forming toxins using Clustal analysis. The alpha toxin (Hla) N-terminus is shown at the top with the boundaries of the NTD neutralizing epitope from alpha toxin (top bar) as mapped by Oscherwitz et al. 2014. Three distinct N-terminal sequences from the monomers comprising the heterodimer LukGH aligned with this region of AT and are designated as LukH (a.a. 28-59), LukH* (a.a. 55-97) and LukG (a.a. 30-60).

[0028] FIG. 4A-4B shows antibody (FIG. 4A) and TNA (FIG. 4B) responses in rabbit serum following immunization with the respective MAP sequences. Horizontal lines represent geometric means.

[0029] FIG. 5 shows the LukGH neutralizing antibody response in the rabbit serum is inhibited by pre-incubation with the LukH and LukH* MAPs but not with the LukG MAP. Shown are ED50 neutralization responses in the TNA from the rabbit sera following pre- incubation of the sera with the indicated MAPs. Error bars represent standard error of the means (SEM).

[0030] FIG. 6 shows a table with synthetic MAPs used for immunization of groups of rabbits to define the fine specificity of the LukH neutralizing epitope(s).

[0031] FIGS. 7A-7B show an antibody (FIG. 7A) and neutralization (FIG. 7B) titers from rabbits immunized with the respective MAP sequences. Error bars represent standard error of the means (SEM).

[0032] FIG. 8 is a table showing the results of peptide inhibition of neutralizing rabbitATTORNEY DOCKET NO. 37759.0626P1 antisera in the TNA. The data identify two distinct linear neutralizing epitopes at the N-terminus of LukH: KEKR (SEQ ID NO:6) and STAPD (SEQ ID NO: 1).

[0033] FIG. 9 shows a diagrammatic of NP123 and NP125 VLPs displaying the STAPD (SEQ ID NO: 1) neutralizing motif.

[0034] FIG. 10 shows an SDS gel demonstrating stages of purification of NP123 and NP125 VLPs. Lane details are shown in the table to right of figure.

[0035] FIGS. 11 A -1 IB show antibody (FIG. 11 A) and neutralization (FIG. 1 IB) titers in the sera of rabbits immunized two times with the respective VLPs, NP123 and NP125, in IF A. In B, an affinity -purified IgG from a group of rabbits immunized with full-length LukGH is shown as a positive control. Horizontal lines are geometric means.

[0036] FIG. 12 shows antisera from rabbits immunized w ith the NP123 VLP neutralizes the cytotoxicity of LukGH-containing supernatant from S. aureus Newman on HL60 cells in vitro. The positive control is a commercially-available, affinity-purified neutralizing IgG obtained from pooled sera from rabbits immunized with full-length LukGH.

[0037] FIG. 13 show s a diagrammatic of VLP128 displaying the KEKR neutralizing motif.

[0038] FIGS. 14A and 14B show the antibody titers (FIG. 14A) in the sera of rabbits immunized two times with the KEKR-displaying VLP 128 in IFA and the toxin neutralizing assay (FIG. 14B). Horizontal line is geometric mean.DETAILED DESCRIPTION

[0039] The disclosed method and compositions may be understood more readily by reference to the following detailed description of particular embodiments and the Example included therein and to the Figures and their previous and following description.

[0040] It is to be understood that the disclosed method and compositions are not limited to specific synthetic methods, specific analytical techniques, or to particular reagents unless otherwise specified, and. as such, may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting.

[0041] Disclosed are materials, compositions, and components that can be used for, can be used in conjunction with, can be used in preparation for, or are products of the disclosed method and compositions. These and other materials are disclosed herein, and it is understood that when combinations, subsets, interactions, groups, etc. of these materials are disclosed that while specific reference of each various individual and collective combinations and permutation of these compounds may not be explicitly disclosed, each is specifically contemplated andATTORNEY DOCKET NO. 37759.0626P1 described herein. For example, if a peptide is disclosed and discussed and a number of modifications that can be made to a number of molecules including the amino acids are discussed, each and every7combination and permutation of the peptide and the modifications that are possible are specifically contemplated unless specifically indicated to the contrary. Thus, if a class of molecules A, B, and C are disclosed as well as a class of molecules D, E. and F and an example of a combination molecule, A-D is disclosed, then even if each is not individually recited, each is individually and collectively contemplated. Thus, is this example, each of the combinations A-E, A-F, B-D, B-E, B-F, C-D, C-E, and C-F are specifically contemplated and should be considered disclosed from disclosure of A, B. and C; D, E, and F; and the example combination A-D. Likewise, any subset or combination of these is also specifically contemplated and disclosed. Thus, for example, the sub-group of A-E, B-F, and C-E are specifically contemplated and should be considered disclosed from disclosure of A, B, and C; D, E, and F; and the example combination A-D. This concept applies to all aspects of this application including, but not limited to, steps in methods of making and using the disclosed compositions. Thus, if there are a variety of additional steps that can be performed it is understood that each of these additional steps can be performed with any specific embodiment or combination of embodiments of the disclosed methods, and that each such combination is specifically contemplated and should be considered disclosed.A. Definitions

[0042] It is understood that the disclosed method and compositions are not limited to the particular methodology, protocols, and reagents described as these may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention which will be limited only by the appended claims.

[0043] It must be noted that as used herein and in the appended claims, the singular forms "a ", "an", and "the" include plural reference unless the context clearly dictates otherwise. Thus, for example, reference to "a peptide" includes a plurality of such peptides, reference to "the peptide” is a reference to one or more peptide and equivalents thereof known to those skilled in the art, and so forth.

[0044] The word “or” as used herein means any one member of a particular list and also includes any combination of members of that list.

[0045] As used herein, the term "therapeutically effective amount" means an amount of a therapeutic, prophylactic, and / or diagnostic agent that is sufficient, when administered to a subject suffering from or susceptible to a disease, disorder, and / or condition, to treat, alleviate,ATTORNEY DOCKET NO. 37759.0626P1 ameliorate, relieve, alleviate symptoms of, prevent, delay onset of, inhibit progression of, reduce severity of, and / or reduce incidence of the disease, disorder, and / or condition.

[0046] As used herein, the term "treating" refers to partially or completely alleviating, ameliorating, relieving, delaying onset of, inhibiting progression of, reducing severity of, and / or reducing incidence of one or more symptoms or features of a particular disease, disorder, and / or condition. For example, "treating" Staphylococcus aureus may refer to inhibiting survival, growth, and / or spread of Staphylococcus aureus. Treatment may be administered to a subject who does not exhibit signs of a disease, disorder, and / or condition and / or to a subject who exhibits only early signs of a disease, disorder, and / or condition for the purpose of decreasing the risk of developing pathology associated with the disease, disorder, and / or condition.

[0047] As used herein, the terms “administering” and “administration” refer to any method of providing a disclosed VLP, peptide, nucleic acid, or composition of the invention to a subject. Such methods are well known to those skilled in the art and include, but are not limited to: oral administration, transdermal administration, administration by inhalation, nasal administration, topical administration, intravaginal administration, ophthalmic administration, intraaural administration, intracerebral administration, rectal administration, sublingual administration, buccal administration, and parenteral administration, including injectable such as intravenous administration, intra-arterial administration, intramuscular administration, and subcutaneous administration. Administration can be continuous or intermittent. In various aspects, a preparation can be administered therapeutically; that is, administered to treat an existing disease or condition. In further various aspects, a preparation can be administered prophylactically; that is, administered for prevention of a disease or condition. In an aspect, the skilled person can determine an efficacious dose, an efficacious schedule, or an efficacious route of administration for a disclosed composition or a disclosed exosome so as to treat a subject.

[0048] As used herein, the term “amino acid sequence” refers to a list of abbreviations, letters, characters or words representing amino acid residues. The amino acid abbreviations used herein are conventional one letter codes for the amino acids and are expressed as follows: A, alanine; C, cysteine; D aspartic acid; E, glutamic acid; F, phenylalanine; G, glycine; H histidine; I isoleucine; K, lysine; L, leucine; M, methionine; N, asparagine; P, proline; Q, glutamine; R, arginine; S, serine; T, threonine; V, valine; W, tryptophan; and Y, tyrosine.

[0049] “Peptide” as used herein refers to any peptide, oligopeptide, polypeptide, gene product, expression product, or protein. A peptide is comprised of consecutive amino acids. The term “peptide” encompasses naturally occurring or synthetic molecules.

[0050] The terms "variant" and "mutant" are used interchangeably herein. As used herein,ATTORNEY DOCKET NO. 37759.0626P1 the term "variant" refers to a modified nucleic acid or peptide / protein which displays the same characteristics when compared to a reference nucleic acid or protein sequence. A variant can be at least 65, 70, 75, 80, 85, 90, 95, or 99 percent homologous to a reference sequence. In some aspects, a reference sequence can be a fragment of one or more of the disclosed sequences. A “variant” can mean a difference in some way from the reference sequence other than just a simple deletion of an N- and / or C-terminal amino acid. A variant can also be a difference in the nucleotide sequence. Variants can also or alternatively include at least one substitution and / or at least one addition; there may also be at least one deletion. Alternatively or in addition, variants can comprise modifications, such as non-natural residues at one or more positions with respect to a reference nucleic acid or protein.

[0051] As used herein, “subject” refers to the target of administration, e.g. an animal. Thus the subject of the disclosed methods can be a vertebrate, such as a mammal. For example, the subject can be a human. The term does not denote a particular age or sex. Subject can be used interchangeably with “individual” or “patient”.

[0052] Ranges may be expressed herein as from "about" one particular value, and / or to "about" another particular value. When such a range is expressed, also specifically contemplated and considered disclosed is the range from the one particular value and / or to the other particular value unless the context specifically indicates otherwise. Similarly, when values are expressed as approximations, by use of the antecedent “about,” it will be understood that the particular value forms another, specifically contemplated embodiment that should be considered disclosed unless the context specifically indicates otherwise. It will be further understood that the endpoints of each of the ranges are significant both in relation to the other endpoint, and independently of the other endpoint unless the context specifically indicates otherwise. Finally, it should be understood that all of the individual values and sub-ranges of values contained within an explicitly disclosed range are also specifically contemplated and should be considered disclosed unless the context specifically indicates otherwise. The foregoing applies regardless of whether in particular cases some or all of these embodiments are explicitly disclosed.

[0053] Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of skill in the art to which the disclosed method and compositions belong. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present method and compositions, the particularly useful methods, devices, and materials are as described. Publications cited herein and the material for which they are cited are hereby specifically incorporated by reference. Nothing herein is to be construed as an admission that the present invention is notATTORNEY DOCKET NO. 37759.0626P1 entitled to antedate such disclosure by virtue of prior invention. No admission is made that any reference constitutes prior art. The discussion of references states what their authors assert, and applicants reserve the right to challenge the accuracy and pertinence of the cited documents. It will be clearly understood that, although a number of publications are referred to herein, such reference does not constitute an admission that any of these documents form part of the common general knowledge in the art.

[0054] Throughout the description and claims of this specification, the word “comprise” and variations of the word, such as “comprising” and “comprises,” means “including but not limited to,” and is not intended to exclude, for example, other additives, components, integers or steps. In particular, in methods stated as comprising one or more steps or operations it is specifically contemplated that each step comprises what is listed (unless that step includes a limiting term such as “consisting of’), meaning that each step is not intended to exclude, for example, other additives, components, integers or steps that are not listed in the step.B. Peptides

[0055] Disclosed are LukH peptides. In some aspects, the LukH peptide is immunogenic. In some aspects, the LukH peptide is from Staphylococcus aureus. LukH is a portion of LukGH. LukGH is commonly referred to as LukAB in the S. aureus literature. As disclosed herein, LukA is equivalent to LukH and LukB is equivalent to LukG.

[0056] In some aspects, the STAPD (SEQ ID NOT) and / or KEKR epitopes of LukH can be immunogenic. Thus, disclosed are peptides comprising the STAPD (SEQ ID NOT) and / or KEKR epitopes alone or in combination with other amino acids.

[0057] Disclosed are peptides comprising or consisting of the amino acid sequence STAPD (SEQ ID NOT), STAPDD (SEQ ID NO:2), TNKDKNSTAPDDIGKNGK (SEQ ID NOT), KDKNSTAPDDIGKNGKI (SEQ ID NO:4), STAPDDIGKNGKITKRT (SEQ ID NO:5), KEKRNVTNKDKNSTAPD (SEQ ID NOTO), KEKR (SEQ ID NO: 6), KDKR (SEQ ID NO: 7), EHVDKSQQKEKRNVTNK (SEQ ID NOT), or KSQQKEKRNTNKDKNS (SEQ ID NOV).

[0058] Disclosed are peptides comprising or consisting essentially of an amino acid sequence at least 90% identical to STAPD (SEQ ID NOT), STAPDD (SEQ ID NOT), TNKDKNSTAPDDIGKNGK (SEQ ID NOT), KDKNSTAPDDIGKNGKI (SEQ ID NOT), STAPDDIGKNGKITKRT (SEQ ID NO:5), KEKRNVTNKDKNSTAPD (SEQ ID NOTO), KEKR (SEQ ID NO:6), KDKR (SEQ ID NOT), EHVDKSQQKEKRNVTNK (SEQ ID NOT), or KSQQKEKRNTNKDKNS (SEQ ID NOV).1. Peptides comprising a STAPD epitope

[0059] Disclosed are peptides, wherein the peptides comprise 5 to 50 amino acids andATTORNEY DOCKET NO. 37759.0626P1 wherein the amino acid sequence comprises the amino acid sequence STAPD. Disclosed are peptides, wherein the peptide consists of 5 to 50 amino acids, wherein the amino acid sequence comprises the amino acid sequence STAPD (SEQ ID NO: 1). In some aspects, the peptides are 5 to 40 amino acids. In some aspects, the peptides are 5 to 30 amino acids. In some aspects, the peptides are 5 to 20 amino acids. In some aspects, the peptides are 5 to 18 amino acids. In some aspects, the peptides are 5 to 10 amino acids. Disclosed are peptides comprising a variant of STAPD (SEQ ID NOT).

[0060] Disclosed are peptides, wherein the peptides comprise 6 to 50 amino acids and wherein the amino acid sequence comprises the amino acid sequence STAPDD. Disclosed are peptides, wherein the peptide consists of 6 to 50 amino acids, wherein the amino acid sequence comprises the amino acid sequence STAPDD (SEQ ID NO: 2). In some aspects, the peptides are 6 to 40 amino acids. In some aspects, the peptides are 6 to 30 amino acids. In some aspects, the peptides are 6 to 20 amino acids. In some aspects, the peptides are 6 to 18 amino acids. In some aspects, the peptides are 6 to 10 amino acids. Disclosed are peptides comprising a variant of STAPDD (SEQ ID NO:2).

[0061] Disclosed are peptides comprising an amino acid sequence that is 90% identical to a portion of LukH. Disclosed are peptides comprising an amino acid sequence that is 60. 65, 70, 75, 80, 85, 90, 95, or 99% identical to a portion of LukH. In some aspects, the portion of LukH comprises the amino acid sequence STAPD (SEQ ID NOT) or STAPDD (SEQ ID NO:2). In some aspects, a peptide comprising an amino acid sequence 80% identical to the amino acid sequence STAPD (SEQ ID NO: 1) comprises a peptide having the amino acid sequence STVPD (SEQ ID NO: 13). In some aspects, a peptide comprising an amino acid sequence 60, 65. 70. 75. 80, 85, 90, 95, or 99% identical to a portion of LukH retains the STAPD (SEQ ID NO: 1) or STAPDD (SEQ ID NO:2) sequence and comprises an amino acid mutation (e.g., substitution, deletion or insertion) somewhere other than the amino acid sequence STAPD (SEQ ID NOT) or STAPDD. In some aspects, the PD of STAPD (SEQ ID NO: 1) or the PDD of STAPDD (SEQ ID NO:2) are retained while one or more of the STA can comprise an amino acid mutation.

[0062] In some aspects, disclosed are peptides comprising amino acids 55 to 77 of full length LukH. In some aspects, disclosed are peptides comprising amino acids 33 to 77 of full length LukH. Also disclosed are peptides comprising an amino acid sequence that is 90% identical to amino acids 33-77 of LukH or amino acids 55-77 of LukH. In some aspects, the disclosed peptides are about 5 to 50 amino acids in length. In some aspects, the disclosed peptides are about 5 to 30 amino acids in length. In some aspects, the disclosed peptides are 5 to 20 amino acids in length. In some aspects, the disclosed peptides are 5 to 18 amino acids inATTORNEY DOCKET NO. 37759.0626P1 length. In some aspects, the disclosed peptides are 5 to 10 amino acids in length.

[0063] In some aspects, the full length LukH peptide sequence is MKNKKRVLIASSLSCAILLLSAATTQANSAHKDSQDQNKKEHVDKSQQKEKRNVTNK DKNSTAPDDIGKNGKITKRTETVYDEKTNILQNLQFDFIDDPTYDKNVLLVKKQGSIHS NLKFESHKEEKNSNWLKYPSEYHVDFQVKRNPKTE1LDQLPKNK1STAKVDSTFSYSSG GKFDSTKGIGRTSSNSYSKTISYNQQNYDTIASGKNNNWHVHWSVIANDLKYGGEVKN RNDELLFYRNTRIATVENPELSFASKYRYPALVRSGFNPEFLTYLSNEKSNEKTQFEVTY TRNQDILKNRPGIHYAPPILEKNKDGQRLIVTYEVDWKNKTVKVVDKYSDDNKPYKEG (SEQ ID NO: 11). Disclosed herein are peptides comprising a portion of LukH comprising at least the STAPD (SEQ ID NO: 1) sequence at amino acids 61-65 of SEQ ID NO: 1. In some aspects, the peptides are about 5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 amino acids in length.

[0064] Disclosed are peptides comprising, consisting essentially of. or consisting of the amino acid sequence STAPD (SEQ ID NO: 1) such as, but not limited to, STAPDD (SEQ ID NO:2), TNKDKNSTAPDDIGKNGK (SEQ ID NO:3), KDKNSTAPDDIGKNGKI (SEQ ID NO:4), STAPDDIGKNGKITKRT (SEQ ID NO:5) or an immunogenic equivalent thereof, wherein the immunogenic equivalent thereof is a peptide comprising, consisting essentially of, or consisting of an amino acid sequence at least 60, 65, 70, 75, 80, 85, 90, 95, or 99% identical to the amino acid sequence STAPDD (SEQ ID NO:2), TNKDKNSTAPDDIGKNGK (SEQ ID NO:3), KDKNSTAPDDIGKNGKI (SEQ ID NO:4), STAPDDIGKNGKITKRT (SEQ ID NO:5).

[0065] Disclosed are peptides, wherein the peptide comprises or consists of 5 to 50 amino acids, wherein the amino acid sequence comprises the amino acid sequence STAPD (SEQ ID NO: 1), and wherein the amino acid sequence comprises an amino acid sequence at least 60, 65, 70, 75, 80, 85, 90, 95, or 99% identical to the amino acid sequence TNKDKNSTAPDDIGKNGK (SEQ ID NO:3).

[0066] Disclosed are peptides, wherein the peptide comprises or consists of 5 to 50 amino acids, wherein the amino acid sequence comprises the amino acid sequence STAPD (SEQ ID NO: 1). and wherein the ammo acid sequence comprises an amino acid sequence at least 60, 65, 70, 75, 80, 85, 90, 95, or 99% identical to the amino acid sequence KDKNSTAPDDIGKNGKI (SEQ ID NO: 4).

[0067] Disclosed are peptides, wherein the peptide comprises or consists of 5 to 50 amino acids, wherein the amino acid sequence comprises the amino acid sequence STAPD (SEQ ID NO: 1), and wherein the amino acid sequence comprises an amino acid sequence at least 60, 65, 70, 75, 80, 85, 90, 95, or 99% identical to the amino acid sequence STAPDDIGKNGKITKRT (SEQ ID NO: 5).ATTORNEY DOCKET NO. 37759.0626P1

[0068] Disclosed are any of the disclosed peptides comprising an amino acid sequence comprising the amino acid sequence STAPD (SEQ ID NO: 1), wherein the amino acid sequence further comprises the amino acid sequence KEKR (SEQ ID NO:6). Thus, in some aspects, the amino acid sequence comprises an amino acid sequence having the amino acid sequence KEKRNVTNKDKNSTAPD (SEQ ID NO: 10) or a variant thereof2. Peptides comprising a KEKR epitope

[0069] Disclosed are peptides, wherein the peptides comprise 4 to 30 amino acids, wherein the amino acid sequence comprises the amino acid sequence KEKR (SEQ ID NO:6). Disclosed are peptides, wherein the peptides consist of 4 to 30 amino acids, wherein the amino acid sequence comprises the amino acid sequence KEKR (SEQ ID NO:6). In some aspects, the peptides are 4 to 50 amino acids. In some aspects, the peptides are 4 to 40 amino acids. In some aspects, the peptides are 4 to 20 amino acids. In some aspects, the peptides are 4 to 17 amino acids. In some aspects, the peptides are 4 to 10 amino acids. Disclosed are peptides comprising a variant of the amino acid sequence KEKR (SEQ ID NO: 6).

[0070] Disclosed are peptides comprising an amino acid sequence that is 90% identical to a portion of LukH. Disclosed are peptides comprising an amino acid sequence that is 60, 65, 70, 75, 80, 85, 90, 95, or 99% identical to a portion of LukH. In some aspects, the portion of LukH comprises the amino acid sequence KEKR (SEQ ID NO:6). In some aspects, a peptide comprising an amino acid sequence 70% identical to the amino acid sequence KEKR (SEQ ID NO:6) comprises a peptide having the sequence of KDKR (SEQ ID NO:7). In some aspects, a peptide comprising an amino acid sequence 60, 65, 70, 75, 80, 85, 90, 95, or 99% identical to a portion of LukH retains the amino acid sequence KEKR (SEQ ID NO:6 and comprises an amino acid mutation (e.g., substitution, deletion or insertion) somewhere other than the amino acid sequence KEKR (SEQ ID NO: 6).

[0071] In some aspects, disclosed are peptides comprising is amino acids 41 to 61 of full length LukH. In some aspects, disclosed are peptides comprising amino acids 33 to 77 of full length LukH. Also disclosed are peptides comprising an amino acid sequence that is 90% identical to amino acids 41-61 of LukH or amino acids 33-77 of LukH. In some aspects, the disclosed peptides are 4 to 50 amino acids in length. In some aspects, the disclosed peptides are 4 to 40 amino acids in length. In some aspects, the disclosed peptides are 4 to 30 amino acids in length. In some aspects, the disclosed peptides are 4 to 20 amino acids in length. In some aspects, the disclosed peptides are 4 to 17 amino acids in length. In some aspects, the disclosed peptides are 4 to 10 amino acids in length.

[0072] Disclosed are peptides comprising a portion of LukH (SEQ ID NO: 11) comprising atATTORNEY DOCKET NO. 37759.0626P1 least the KEKR (SEQ ID N0:6) sequence at amino acids 49-52 of SEQ ID NO: 11. In some aspects, the peptides are about 4 to 50 amino acids in length.

[0073] Disclosed are peptides comprising, consisting essentially of, or consisting of the amino acid sequence KEKR (SEQ ID NO:6) such as, but not limited to, EHVDKSQQKEKRNVTNK (SEQ ID NOV), KSQQKEKRNTNKDKNS (SEQ ID NOV), or an immunogenic equivalent thereof, wherein the immunogenic equivalent thereof is a peptide comprising, consisting essentially of, or consisting of an amino acid sequence at least 60, 65, 70, 75, 80, 85, 90, 95, or 99% identical to the amino acid sequence EHVDKSQQKEKRNVTNK (SEQ ID NOV) or KSQQKEKRNTNKDKNS (SEQ ID NOV).

[0074] Disclosed are peptides, wherein the peptide comprises or consists of 4 to 30 amino acids, wherein the amino acid sequence comprises the amino acid sequence KEKR (SEQ ID NO:6), and wherein the amino acid sequence comprises an amino acid sequence at least 60, 65, 70, 75, 80, 85, 90, 95, or 99% identical to the amino acid sequence EHVDKSQQKEKRNVTNK (SEQ ID NO: 8).

[0075] Disclosed are peptides, wherein the peptide comprises or consists of 4 to 30 amino acids, wherein the amino acid sequence comprises the amino acid sequence KEKR (SEQ ID NO:6), and wherein the amino acid sequence comprises an amino acid sequence at least 60, 65, 70, 75, 80, 85, 90, 95, or 99% identical to the amino acid sequence KSQQKEKRNTNKDKNS (SEQ ID NO: 9)

[0076] In some aspects, any amino acid within the amino acid sequence EHVDKSQQKEKRNVTNK (SEQ ID NO: 8) or KSQQKEKRNTNKDKNS (SEQ ID NOV) can be mutated except for the KEKR (SEQ ID NOV) sequence. In some aspects, the E or R of the KEKR (SEQ ID NOV) sequence can be mutated in the disclosed sequences while the lysines are retained. In some aspects, the E can be mutated to D resulting in a KDKR (SEQ ID NOV) sequence wherein the lysines are retained and the R can be retained or also mutated.

[0077] Disclosed are any of the disclosed peptides comprising an amino acid sequence comprising the amino acid sequence KEKR (SEQ ID NOV), wherein the amino acid sequence further comprises the amino acid sequence STAPD (SEQ ID NO:1) or STAPDD (SEQ ID NOV). Thus, in some aspects, the amino acid sequence comprises an amino acid sequence having the amino acid sequence KEKJ<NVTNKDKNSTAPD (SEQ ID NO: 10) or a variant thereof.3. Helper T cell epitope

[0078] In some aspects, the disclosed peptides can further comprise a helper T cell epitope. In some aspects, the presence of a helper T cell epitope can help trigger a B cell response, andATTORNEY DOCKET NO. 37759.0626P1 thus, help in eliciting an antibody response.

[0079] Disclosed are polypeptides comprising a first peptide and a second peptide, wherein the first peptide is any one of peptides comprising the sequence STAPD (SEQ ID NO: 1) or KEKR (SEQ ID NO:6) disclosed herein, wherein the second peptide is a helper T cell epitope.

[0080] In some aspects, the first peptide is on the N-terminal or C-terminal end of the helper T cell epitope.

[0081] In some aspects, the helper T cell epitope can be any known helper T cell epitope. In some aspects, the helper T cell epitope can be from Plasmodium falciparum or tetanus toxin. In some aspects, the helper T cell epitope can be T*. a helper T cell epitope tmm Plasmodium falciparumi. In some aspects, the helper T cell epitope can be P30, a helper T cell epitope from tetanus toxin.C. Virus-Like Particles

[0082] In some aspects, one or more of the disclosed peptides can be displayed on virus-like particles, for example, as part of a recombinantly produced viral capsid protein. Disclosed are modified proteins, wherein a modified protein is any protein that plays a role in forming a virus like particle (VLP) that has been modified to include the amino acid sequence of one or more of the disclosed peptides. In some aspects, a modified protein can be a modified viral capsid protein.

[0083] As shown in FIG. 9, a VLP assembled particle comprises many VLP monomers assembled together into a nanoparticle. A VLP monomer can also be referred to as a modified capsid protein monomer and each monomer displays sequences which are presented on the surface of the assembled VLP nanoparticle. For example, each VLP monomer can display a sequence, such as KEKR (SEQ ID NO:6) or STAPD (SEQ ID NO: 1), and these sequences are presented on the surface of the assembled VLP nanoparticle.

[0084] Disclosed are modified viral capsid proteins comprising a portion of LukH, wherein the portion of LukH comprises the amino acid sequence STAPD (SEQ ID NO: 1 ), KEKR (SEQ ID NO:6), or both.

[0085] In some aspects, the modified viral capsid protein is a modified w oodchuck hepatitis virus capsid protein.

[0086] In some aspects, the portion of LukH is inserted within the modified viral capsid protein. In some aspects, the portion of LukH can be inserted in the N-terminal end, C-terminal end, or in the middle of a capsid protein. Thus, in some aspects, the portion of LukH is expressed as part of the capsid protein and not simply conjugated to the capsid protein.

[0087] In some aspects, the disclosed modified capsid proteins are present as virus-likeATTORNEY DOCKET NO. 37759.0626P1 particles (VLP). In some aspects, the VLP comprises 240 modified capsid protein monomers on the surface of the VLP assembled nanoparticle. In some aspects, the VLP comprises 50-240 modified capsid protein monomers on the surface of the VLP assembled nanoparticle.

[0088] In some aspects, the disclosed peptides can be inserted within an amino acid sequence of a polypeptide capable of forming a virus like particle. In some aspects, a portion of LukH comprising the amino acid sequence STAPD (SEQ ID NO: 1), KEKR (SEQ ID NO:6) or both can be inserted within an amino acid sequence of a polypeptide capable of forming a virus like particle. In some aspects, an amino acid sequence comprising STAPD (SEQ ID NO: 1) and an amino acid sequence comprising KEKR (SEQ ID NO:6) can be inserted separately within an amino acid sequence of a polypeptide capable of forming a virus like particle. In some aspects, a portion of LukH comprising the amino acid sequence STAPDD (SEQ ID NO:2) can be inserted within an amino acid sequence of a polypeptide capable of forming a virus like particle. In some aspects, a portion of LukH comprising one or more of the amino acid sequences described herein can be inserted within an amino acid sequence of a polypeptide capable of forming a virus like particle.

[0089] In some aspects, when two or more of the disclosed peptides are each individually inserted in a polypeptide capable of forming a VLP. the VLP can include one or all of the peptides. In some aspects, when two or more of the disclosed peptides are each individually inserted in a polypeptide capable of forming a VLP, the VLP can include all of the peptides. Therefore, in some aspects, the VLPs can be distinct for each of the disclosed peptides or can comprise more than one of the antigenic peptides.

[0090] In some aspects, a VLP assembled particle can comprise 50-240 VLP monomers wherein each of the VLP monomers is identical. In some embodiments, the peptide comprising the amino acid sequence STAPD (SEQ ID NO: 1) and a peptide comprising the amino acid sequence KEKR (SEQ ID NO:6) are in distinct virus like particles, such that the composition can comprise one VLP with polypeptides in which the peptide comprising the amino acid sequence STAPD (SEQ ID NO: 1) is inserted and a second VLP with polypeptides in which the peptide comprising the amino acid sequence KEKR (SEQ ID NO:6) is inserted.

[0091] Examples of polypeptide capable of forming a virus like particle (VLP) include, but are not limited to, polypeptides of QP, MS2, PP7, AP205 and other bacteriophage coat proteins and the capsid and core proteins / polypeptides of a wide variety of virus families including Parvoviridae (e.g., adeno-associated virus), Retroviridae (e.g., HIV), Flaviviridae (e.g., Hepatitis C virus), Hepadnaviridae (e.g., Hepatitis B virus), Paramyxoviridae (e.g., measles virus), Paramyxoviridae (e.g., Nipah), Togaviridae (e.g., Sindbis virus), Picomaviridae (e.g., foot-and-ATTORNEY DOCKET NO. 37759.0626P1 mouth disease virus), Caliciviridae (e.g.. Nowalk virus), Bromoviridae (e.g., cowpea mosaic virus), polyomaviridae (e.g., JC virus), and Papillomaviridae (HPV).

[0092] In some aspects, the VLPs can be synthesized chemically or through a biological process, include a variety' of cell culture systems, including bacterial, mammalian, insect, yeast, and plant cells. In certain embodiments, the VLP can comprise recombinant polypeptides of any of the virus known to form a VLP to create a recombinant VLP. The virus-like particle can further comprise, or alternatively consist of, one or more fragments of such polypeptides, as well as variants of such polypeptides.

[0093] Variants of polypeptides can share, for example, at least 80%. 85%. 90%. 95%. 97%. or 99% identity' at the amino acid level with their wild-type counterparts.

[0094] In some aspects, the polypeptide capable of forming a virus like particle is a woodchuck hepatitis DNA virus core antigen. The basic subunit of the core particle is a 21 kDa polypeptide monomer that spontaneously assembles into a 240-subunit structure of about 34 nm in diameter. In some aspects, any of the know n hepatitis virus core proteins sequences may be appropriate for use with peptide described herein, such as those described in U.S. Patent No. 7,883,843, incorporated herein in its entirety' by reference. In some aspects, the hepatitis virus is woodchuck hepatitis DNA virus. In some embodiments, the amino acid sequence of the woodchuck hepatitis DNA virus core antigen may comprise an amino acid sequence having at least 70% identity (e.g., at 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% identity ) to Woodchuck core proteinMDIDPYKEFGSSYQLLNFLPLDFFPDLNALVDTATALYEEELTGREHCSPHHTAIRQALV CWDELTKLIAWMSSNITSEQVRTIIVNHVNDTWGLKVRQSLWFHLSCLTFGQHTVQEFL VSFGVWIRTPAPYRPPNAPILSTLPEHTVIRRRGGARASRSPRRRTPSPRRRRSQSPRRRR SQSPSANC (SEQ ID NO: 12).

[0095] In some aspects, in order to not disrupt self-assembly of the core antigen VLP, the disclosed peptides can be inserted inside the loop region for the hepatitis DNA virus core protein sequence, such as, for example, ammo acid residues 76. 77. 78. 81. and / or 82 of woodchuck hepatitis DNA virus core antigen. In certain embodiments, the insertion into the woodchuck hepatitis DNA virus core antigen may be at position 78 of SEQ ID NO: 12.

[0096] In some aspects, the disclosed peptides can be present at the N-terminus or C- terminus of the VLP monomer. In some aspects, the disclosed peptides can be present anywhere within or at the ends of the VLP monomer.

[0097] In some aspects, a VLP monomer can comprise one peptide (e.g. KEKR) inserted inside the loop region for the hepatitis DNA virus core protein sequence, for example at positionATTORNEY DOCKET NO. 37759.0626P178 of woodchuck hepatitis DNA virus core antigen, and one peptide (e.g. STAPD) at the N- terminus (or C-terminus). Thus, in this scenario, all of the VLP monomers in the assembled VLPs comprise the two peptides.

[0098] Thus, in some aspects, disclosed are virus-like particles (VLPs) comprising any one of the modified proteins disclosed herein used to produce VLPs. In some aspects, the VLPs comprise at least 50 of the modified viral capsid proteins on the surface of the VLP.D. Nucleic Acid Sequences

[0099] Disclosed are nucleic acid sequences or constructs comprising a nucleic acid sequence encoding one or more of the disclosed peptides. Disclosed are nucleic acid sequences encoding an antigenic LukH peptide. Disclosed are nucleic acid sequences or constructs comprising a nucleic acid sequence capable of encoding STAPD (SEQ ID NO:1), STAPDD (SEQ ID NO:2), TNKDKNSTAPDDIGKNGK (SEQ ID NOT), KDKNSTAPDDIGKNGKI (SEQ ID NO:4), STAPDDIGKNGKITKRT (SEQ ID NOT), KEKRNVTNKDKNSTAPD (SEQ ID NOTO), KEKR (SEQ ID NO:6), KDKR (SEQ ID NOT), EHVDKSQQKEKRNVTNK (SEQ ID NO:8), or KSQQKEKRNTNKDKNS (SEQ ID NOV), or a functional vanant thereof.

[0100] In some aspects, the nucleic acids capable of encoding STAPD (SEQ ID NOT), TNKDKNSTAPDDIGKNGK (SEQ ID NO: 6), KDKNSTAPDDIGKNGKI (SEQ ID NO: 4), STAPDDIGKNGKITKRT (SEQ ID NOT), KEKRNVTNKDKNSTAPD (SEQ ID NOTO), KEKR (SEQ ID NO:6), KDKR (SEQ ID NOT), EHVDKSQQKEKRNVTNK (SEQ ID NO: 8), or KSQQKEKRNTNKDKNS (SEQ ID NOV), or a functional variant thereof can be inserted within a nucleic acid sequence encoding the amino acid sequence of a polypeptide capable of forming a virus like particle. In some embodiments, the polypeptide capable of forming a virus like particle is a woodchuck hepatitis DNA virus core antigen. In certain embodiments, the amino acid sequence of the woodchuck hepatitis DNA virus core antigen may comprise an amino acid sequence having at least 70% identity7(e.g., at 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% identity7) to SEQ ID NO: 12. In exemplary embodiments, the insertion into the woodchuck hepatitis DNA virus core antigen may be at position 78 of the amino acid sequence SEQ ID NO: 12.

[0101] In some aspects, the disclosed nucleic acid sequences can be DNA or RNA based.E. Vectors

[0102] Disclosed are vectors comprising one or more of the disclosed nucleic acid sequences. Disclosed are vectors comprising a nucleic acid sequence that encodes any of the disclosed peptides. In some aspects, the vector can be selected from the group consisting of a DNA, a RNA, a plasmid, and a viral vector. In some instances, the vector can comprise aATTORNEY DOCKET NO. 37759.0626P1 promoter.

[0103] In some aspects, the nucleic acid sequences disclosed herein can be introduced into an expression vector, such that the expression vector comprises a promoter and the polynucleotides encoding the peptides or polypeptides described herein. The expression vector may allow expression of the peptides or polypeptides in a suitable expression system using techniques well known in the art, followed by isolation or purification of the expressed peptide or polypeptide of interest. A variety of bacterial, yeast, plant, mammalian, and insect expression systems are available in the art and any such expression system can be used. Alternatively, a polynucleotide encoding a peptide of the invention can be translated in a cell-free translation system. Thus also disclosed are expression vectors comprising one or more of the disclosed peptides or modified protein sequences. For example, an expression vector can be a VLP expressing a peptide consisting of or comprising the amino acid sequence STAPD (SEQ ID NO:1), STAPDD (SEQ ID NO:2), TNKDKNSTAPDDIGKNGK (SEQ IDN0:3), KDKNSTAPDDIGKNGKI (SEQ ID NO:4), STAPDDIGKNGKITKRT (SEQ ID NO:5), KEKRNVTNKDKNSTAPD (SEQ ID NO: 10), KEKR (SEQ ID NO: 6), KDKR (SEQ ID NO: 7), EHVDKSQQKEKRNVTNK (SEQ ID NO:8), or KSQQKEKRNTNKDKNS (SEQ ID NOV). In some aspects, the expression vector expresses one or more of the disclosed peptides as part of one of its native proteins. For example, a peptide consisting of or comprising the amino acid sequence STAPD (SEQ ID NO: 1), STAPDD, TNKDKNSTAPDDIGKNGK, KDKNSTAPDDIGKNGKI, STAPDDIGKNGKITKRT, KEKRNVTNKDKNSTAPD, KEKR, KDKR, EHVDKSQQKEKRNVTNK, or KSQQKEKRNTNKDKNS can be expressed with a capsid protein, thus expression of the capsid protein also expresses the disclosed peptide.

[0104] In some aspects, the vector can be an expression vector. The term "expression vector" includes any vector, (e.g., a plasmid, cosmid or phage chromosome) containing a gene construct in a form suitable for expression by a cell (e.g., linked to a transcriptional control element). "Plasmid" and "vector" can be used interchangeably, as a plasmid is a commonly used form of vector. Moreover, the invention is intended to include other vectors which serve equivalent functions.

[0105] In some aspects, the vector can be a viral vector. For example, the viral vector can be a lentiviral vector. In some aspects, the vector can be a non-viral vector, such as a DNA based vector. i. Viral and Non- Viral Vectors

[0106] There are a number of compositions and methods which can be used to deliver the disclosed nucleic acids to cells, either in vitro or in vivo. These methods and compositions canATTORNEY DOCKET NO. 37759.0626P1 largely be broken down into two classes: viral based delivery systems and non-viral based delivery systems. For example, the nucleic acids can be delivered through a number of direct delivery' systems such as, electroporation, lipofection, calcium phosphate precipitation, plasmids, viral vectors, viral nucleic acids, phage nucleic acids, phages, cosmids. or via transfer of genetic material in cells or carriers such as cationic liposomes. Appropriate means for transfection, including viral vectors, chemical transfectants, or physico-mechanical methods such as electroporation and direct diffusion of DNA, are described by, for example, Wolff, J. A., et al., Science, 247, 1465-1468, (1990); and Wolff, J. A. Nature, 352. 815-818. (1991). Such methods are well known in the art and readily adaptable for use with the compositions and methods described herein. In certain cases, the methods will be modified to specifically function with large DNA molecules. Further, these methods can be used to target certain diseases and cell populations by using the targeting characteristics of the carrier.

[0107] Expression vectors can be any nucleotide construction used to deliver genes or gene fragments into cells (e.g., a plasmid), or as part of a general strategy to deliver genes or gene fragments, e.g., as part of recombinant retrovirus or adenovirus (Ram et al. Cancer Res. 53:83- 88, (1993)). For example, disclosed herein are expression vectors comprising a nucleic acid sequence capable of encoding one or more of the disclosed CAR polypeptides.

[0108] The ‘"control elements” present in an expression vector are those non-translated regions of the vector-enhancers, promoters, 5’ and 3’ untranslated regions— which interact with host cellular proteins to carry out transcription and translation. Such elements may vary in their strength and specificity'. Depending on the vector system and host utilized, any number of suitable transcription and translation elements, including constitutive and inducible promoters, may be used. For example, when cloning in bacterial systems, inducible promoters such as the hybrid lacZ promoter of the pBLUESCRIPT phagemid (Stratagene, La Jolla, Calif) or pSPORTl plasmid (Gibco BRL, Gaithersburg, Md.) and the like may be used. If it is necessary' to generate a cell line that contains multiple copies of the sequence encoding a polypeptide, vectors based on SV40 or EBV may be advantageously used with an appropriate selectable marker.

[0109] Enhancer generally refers to a sequence of DNA that functions at no fixed distance from the transcription start site and can be either 5’ (Laimins, L. et al., Proc. Natl. Acad. Sci. 78: 993 (1981)) or 3T(Lusky, M.L., et al.. Mol. Cell Bio. 3: 1108 (1983)) to the transcription unit. Furthermore, enhancers can be within an intron (Banerji, J.L. et al., Cell 33: 729 (1983)) as well as within the coding sequence itself (Osborne, T.F., et al.. Mol. Cell Bio. 4: 1293 (1984)). They are usually between 10 and 300 bp in length, and they function in cis. Enhancers function toATTORNEY DOCKET NO. 37759.0626P1 increase transcription from nearby promoters. Enhancers also often contain response elements that mediate the regulation of transcription. Promoters can also contain response elements that mediate the regulation of transcription. Enhancers often determine the regulation of expression of a gene. While many enhancer sequences are now known from mammalian genes (globin, elastase, albumin, a-fetoprotein and insulin), typically one will use an enhancer from a eukaryotic cell virus for general expression. Preferred examples are the SV40 enhancer on the late side of the replication origin (bp 100-270), the cytomegalovirus early promoter enhancer, the polyoma enhancer on the late side of the replication origin, and adenovirus enhancers.

[0110] The promoter or enhancer may be specifically activated either by light or specific chemical events which trigger their function. Systems can be regulated by reagents such as tetracycline and dexamethasone. There are also ways to enhance viral vector gene expression by exposure to irradiation, such as gamma irradiation, or alkylating chemotherapy drugs.

[0111] Optionally, the promoter or enhancer region can act as a constitutive promoter or enhancer to maximize expression of the polynucleotides of the invention. In certain constructs the promoter or enhancer region be active in all eukaryotic cell types, even if it is only expressed in a particular type of cell at a particular time.

[0112] Expression vectors used in eukaryotic host cells (yeast, fungi, insect, plant, animal, human or nucleated cells) may also contain sequences necessary for the termination of transcription which may affect mRNA expression. These regions are transcribed as poly adenylated segments in the untranslated portion of the mRNA encoding tissue factor protein. The 3’ untranslated regions also include transcription termination sites. It is preferred that the transcription unit also contains a polyadenylation region. One benefit of this region is that it increases the likelihood that the transcribed unit will be processed and transported like mRNA. The identification and use of poly adenylation signals in expression constructs is well established. It is preferred that homologous polyadenylation signals be used in the transgene constructs. In certain transcription units, the polyadenylation region is derived from the SV40 early poly adenylation signal and consists of about 400 bases.

[0113] The expression vectors can include a nucleic acid sequence encoding a marker product. This marker product can be used to determine if the gene has been delivered to the cell and once delivered is being expressed. Marker genes can include, but are not limited to the E. coli lacZ gene, which encodes B-galactosidase, and the gene encoding the green fluorescent protein.

[0114] In some embodiments the marker may be a selectable marker. Examples of suitable selectable markers for mammalian cells are dihydrofolate reductase (DHFR). thymidine kinase,ATTORNEY DOCKET NO. 37759.0626P1 neomycin, neomycin analog G418, hydromycin, and puromycin. When such selectable markers are successfully transferred into a mammalian host cell, the transformed mammalian host cell can survive if placed under selective pressure. There are two widely used distinct categories of selective regimes. The first category is based on a cell's metabolism and the use of a mutant cell line which lacks the ability to grow independent of a supplemented media. Two examples are CHO DHFR-cells and mouse LTK-cells. These cells lack the ability to grow without the addition of such nutrients as thymidine or hypoxanthine. Because these cells lack certain genes necessary for a complete nucleotide synthesis pathway, they cannot survive unless the missing nucleotides are provided in a supplemented media. An alternative to supplementing the media is to introduce an intact DHFR or TK gene into cells lacking the respective genes, thus altering their growth requirements. Individual cells which were not transformed with the DHFR or TK gene will not be capable of survival in non-supplemented media.

[0115] Another type of selection that can be used with the composition and methods disclosed herein is dominant selection which refers to a selection scheme used in any cell type and does not require the use of a mutant cell line. These schemes typically use a drug to arrest grow th of a host cell. Those cells which have a novel gene would express a protein conveying drug resistance and would survive the selection. Examples of such dominant selection use the drugs neomycin, (Southern P. and Berg. P., J. Molec. Appl. Genet. 1: 327 (1982)). mycophenolic acid, (Mulligan, R.C. and Berg, P. Science 209: 1422 (1980)) or hygromycin, (Sugden, B. et al., Mol. Cell. Biol. 5: 410-413 (1985)). The three examples employ bacterial genes under eukary otic control to convey resistance to the appropriate drug G418 or neomycin (geneticin), xgpt (mycophenolic acid) or hygromycin. respectively. Others include the neomycin analog G418 and puramycin.

[0116] As used herein, plasmid or viral vectors are agents that transport the disclosed nucleic acids, such as a nucleic acid sequence capable of encoding one or more of the disclosed peptides into the cell without degradation and include a promoter yielding expression of the gene in the cells into which it is delivered. In some embodiments the nucleic acid sequences disclosed herein are derived from either a virus or a retrovirus. Viral vectors are, for example, Adenovirus, Adeno-associated virus, Herpes virus, Vaccinia vims, Polio virus, AIDS vims, neuronal trophic virus, Sindbis and other RNA viruses, including these vimses with the HIV backbone. Also preferred are any viral families which share the properties of these vimses which make them suitable for use as vectors. Retroviruses include Murine Maloney Leukemia vims, MMLV, and retrovimses that express the desirable properties of MMLV as a vector. Retroviral vectors are able to carry a larger genetic payload, i.e., a transgene or marker gene,ATTORNEY DOCKET NO. 37759.0626P1 than other viral vectors, and for this reason are a commonly used vector. However, they are not as useful in non-proliferating cells. Adenovirus vectors are relatively stable and easy to work with, have high titers, and can be delivered in aerosol formulation, and can transfect nondividing cells. Pox viral vectors are large and have several sites for inserting genes, they are thermostable and can be stored at room temperature. A preferred embodiment is a viral vector which has been engineered so as to suppress the immune response of the host organism, elicited by the viral antigens. Preferred vectors of this type will cany coding regions for Interleukin 8 or 10.

[0117] Viral vectors can have higher transaction abilities (z.e., ability to introduce genes) than chemical or physical methods of introducing genes into cells. Typically, viral vectors contain, nonstructural early genes, structural late genes, an RNA polymerase III transcript, inverted terminal repeats necessary' for replication and encapsidation, and promoters to control the transcription and replication of the viral genome. When engineered as vectors, viruses ty pically have one or more of the early genes removed and a gene or gene / promoter cassette is inserted into the viral genome in place of the removed viral DNA. Constructs of this type can carry' up to about 8 kb of foreign genetic material. The necessary' functions of the removed early genes are typically supplied by cell lines which have been engineered to express the gene products of the early genes in trans.

[0118] Retroviral vectors, in general, are described by Verma, I.M., Retroviral vectors for gene transfer. In Microbiology7, Amer. Soc. for Microbiology7, pp. 229-232, Washington, (1985), which is hereby incorporated by reference in its entirety. Examples of methods for using retroviral vectors for gene therapy are described in U.S. Patent Nos. 4,868,116 and 4,980,286; PCT applications WO 90 / 02806 and WO 89 / 07136; and Mulligan, (Science 260:926-932 (1993)); the teachings of which are incorporated herein by reference in their entirety for their teaching of methods for using retroviral vectors for gene therapy.

[0119] A retrovirus is essentially a package which has packed into it nucleic acid cargo. The nucleic acid cargo carries with it a packaging signal, which ensures that the replicated daughter molecules will be efficiently packaged within the package coat. In addition to the package signal, there are a number of molecules which are needed in cis, for the replication, and packaging of the replicated virus. Typically a retroviral genome contains the gag, pol, and env genes which are involved in the making of the protein coat. It is the gag. pol, and env genes which are typically replaced by the foreign DNA that it is to be transferred to the target cell. Retrovirus vectors typically contain a packaging signal for incorporation into the package coat, a sequence which signals the start of the gag transcription unit, elements necessary for reverseATTORNEY DOCKET NO. 37759.0626P1 transcription, including a primer binding site to bind the tRNA primer of reverse transcription, terminal repeat sequences that guide the switch of RNA strands during DNA synthesis, a purine rich sequence 5' to the 3' LTR that serves as the priming site for the synthesis of the second strand of DNA synthesis, and specific sequences near the ends of the LTRs that enable the insertion of the DNA state of the retrovirus to insert into the host genome. This amount of nucleic acid is sufficient for the delivery of a one to many genes depending on the size of each transcript. It is preferable to include either positive or negative selectable markers along with other genes in the insert.

[0120] Since the replication machinery and packaging proteins in most retroviral vectors have been removed (gag, pol, and env), the vectors are typically generated by placing them into a packaging cell line. A packaging cell line is a cell line which has been transfected or transformed with a retrovirus that contains the replication and packaging machinery' but lacks any packaging signal. When the vector carrying the DNA of choice is transfected into these cell lines, the vector containing the gene of interest is replicated and packaged into new retroviral particles, by the machinery provided in cis by the helper cell. The genomes for the machinery are not packaged because they lack the necessary' signals.

[0121] The construction of replication-defective adenoviruses has been described (Berkner et al., J. Virology' 61: 1213-1220 (1987); Massie et al.. Mol. Cell. Biol. 6:2872-2883 (1986); Haj- Ahmad et al., J. Virology 57:267-274 (1986); Davidson et al., J. Virology 61: 1226-1239 (1987); Zhang “Generation and identification of recombinant adenovirus by liposome-mediated transfection and PCR analysis'’ BioTechniques 15:868-872 (1993)). The benefit of the use of these viruses as vectors is that they are limited in the extent to which they can spread to other cell types, since they can replicate within an initial infected cell but are unable to form new infectious viral particles. Recombinant adenoviruses have been shown to achieve high efficiency gene transfer after direct, in vivo delivery to airway epithelium, hepatocytes, vascular endothelium, CNS parenchyma and a number of other tissue sites (Morsy, J. Clin. Invest. 92: 1580-1586 (1993); Kirshenbaum, J. Clin. Invest. 92:381-387 (1993); Roessler, J. Clin. Invest. 92: 1085-1092 (1993); Moullier, Nature Genetics 4:154-159 (1993); La Salle, Science 259:988- 990 (1993); Gomez-Foix, J. Biol. Chem. 267:25129-25134 (1992); Rich, Human Gene Therapy 4:461-476 (1993); Zabner, Nature Genetics 6:75-83 (1994); Guzman, Circulation Research 73: 1201-1207 (1993); Bout, Human Gene Therapy 5:3-10 (1994); Zabner. Cell 75:207-216 (1993); Caillaud, Eur. J. Neuroscience 5: 1287-1291 (1993); and Ragot, J. Gen. Virology 74:501-507 (1993)) the teachings of which are incorporated herein by reference in their entirety for their teaching of methods for using retroviral vectors for gene therapy. RecombinantATTORNEY DOCKET NO. 37759.0626P1 adenoviruses achieve gene transduction by binding to specific cell surface receptors, after which the virus is internalized by receptor-mediated endocytosis, in the same manner as wild type or replication-defective adenovirus (Chardonnet and Dales, Virology740:462-477 (1970); Brown and Burlingham, J. Virology 12:386-396 (1973); Svensson and Persson, J. Virology 55:442-449 (1985); Seth, etal., J. Virol. 51:650-655 (1984); Seth, et al.. Mol. Cell. Biol., 4: 1528-1533 (1984); Varga et al., J. Virology 65:6061-6070 (1991); Wickham et al. , Cell 73:309-319 (1993)).

[0122] A viral vector can be one based on an adenovirus which has had the El gene removed and these virions are generated in a cell line such as the human 293 cell line. Optionally, both the El and E3 genes are removed from the adenovirus genome.

[0123] Another type of viral vector that can be used to introduce the polynucleotides of the invention into a cell is based on an adeno-associated virus (AAV). This defective parvovirus is a preferred vector because it can infect many cell types and is nonpathogenic to humans. AAV type vectors can transport about 4 to 5 kb and wild type AAV is known to stably insert into chromosome 19. Vectors which contain this site specific integration property are preferred. An especially preferred embodiment of this ty pe of vector is the P4. 1 C vector produced by Avigen, San Francisco, CA, which can contain the herpes simplex virus thymidine kinase gene, HSV-tk, or a marker gene, such as the gene encoding the green fluorescent protein, GFP.

[0124] In another type of AAV virus, the AAV contains a pair of inverted terminal repeats (ITRs) which flank at least one cassette containing a promoter which directs cell-specific expression operably linked to a heterologous gene. Heterologous in this context refers to any nucleotide sequence or gene which is not native to the AAV or Bl 9 parvovirus. Typically the AAV and Bl 9 coding regions have been deleted, resulting in a safe, noncytotoxic vector. The AAV ITRs, or modifications thereof, confer infectivity and site-specific integration, but not cytotoxicity7, and the promoter directs cell-specific expression. United States Patent No. 6,261,834 is herein incorporated by reference in its entirety for material related to the AAV vector.

[0125] The inserted genes in viral and retroviral vectors usually contain promoters, or enhancers to help control the expression of the desired gene product. A promoter is generally a sequence or sequences of DNA that function when in a relatively fixed location in regard to the transcription start site. A promoter contains core elements required for basic interaction of RNA polymerase and transcription factors, and may contain upstream elements and response elements.

[0126] Other useful systems include, for example, replicating and host-restricted non-ATTORNEY DOCKET NO. 37759.0626P1 replicating vaccinia virus vectors. In addition, the disclosed nucleic acid sequences can be delivered to a target cell in a non-nucleic acid based system. For example, the disclosed polynucleotides can be delivered through electroporation, or through lipofection, or through calcium phosphate precipitation. The delivery’ mechanism chosen will depend in part on the type of cell targeted and whether the delivery is occurring for example in vivo or in vitro.

[0127] Thus, the compositions can comprise, in addition to the disclosed expression vectors, lipids such as liposomes, such as cationic liposomes (e.g., DOTMA, DOPE, DC-cholesterol) or anionic liposomes. Liposomes can further comprise proteins to facilitate targeting a particular cell, if desired. Administration of a composition comprising a peptide and a cationic liposome can be administered to the blood, to a target organ, or inhaled into the respiratory tract to target cells of the respiratory tract. For example, a composition comprising a peptide or nucleic acid sequence described herein and a cationic liposome can be administered to a subjects lung cells. Regarding liposomes, see, e.g., Brigham et al. Am. J. Resp. Cell. Mol. Biol. 1 :95-100 (1989); Feigner et al. Proc. Natl. Acad. Sci USA 84:7413-7417 (1987); U.S. Patent No. 4,897,355. Furthermore, the compound can be administered as a component of a microcapsule that can be targeted to specific cell types, such as macrophages, or where the diffusion of the compound or delivery' of the compound from the microcapsule is designed for a specific rate or dosage.F. Compositions

[0128] Disclosed are compositions comprising one or more of the disclosed peptides, modified proteins, nucleic acids, vectors, or VLPs. In some instances, disclosed are compositions comprising a peptide comprising / consisting essentially of the amino acid sequence STAPD (SEQ ID NO: 1). STAPDD (SEQ ID NO:2), TNKDKNSTAPDDIGKNGK (SEQ ID NO:3), KDKNSTAPDDIGKNGKI (SEQ ID NO:4), STAPDDIGKNGKITKRT (SEQ ID NO:5) or a peptide equivalent thereof, wherein the peptide equivalent thereof is a peptide comprising / consisting essentially of an amino acid sequence at least 90% identical to the amino acid sequence STAPD (SEQ ID NO: 1), STAPDD (SEQ ID NO:2).TNKDKNSTAPDDIGKNGK (SEQ ID NO:3), KDKNSTAPDDIGKNGKI (SEQ ID NO:4). STAPDDIGKNGKITKRT (SEQ ID NO:5). In some aspects, the peptide can be referred to as an immunogen. In some aspects, an immunogen is a substance that can induce an immune response.

[0129] Disclosed are compositions comprising an peptide comprising / consisting essentially of the amino acid sequence KEKR (SEQ ID NO: 6) such as, but not limited to, the amino acid sequence EHVDKSQQKEKRNVTNK (SEQ ID NO: 8), KSQQKEKRNTNKDKNS (SEQ ID NO:9), or a peptide equivalent thereof, wherein the peptide equivalent thereof is a peptideATTORNEY DOCKET NO. 37759.0626P1 comprising, consisting essentially of, or consisting of an amino acid sequence at least 60, 65, 70, 75, 80, 85, 90, 95, or 99% identical to the amino acid sequence EHVDKSQQKEKRNVTNK (SEQ ID NO:8) or KSQQKEKRNTNKDKNS (SEQ ID NO:9).

[0130] In some aspects, the composition can be a vaccine. In some aspects, composition and vaccine can be used interchangeably. Thus, disclosed are compositions that can be used to generate an immune response, therefore can be referred to as immunogenic compositions.

[0131] In some aspects, the compositions can further comprising an adjuvant or immunostimulant. Adjuvants and immunostimulants are compounds that either directly or indirectly stimulate the immune system’s response to a co- administered antigen. Suitable adjuvants are commercially available as, for example, Freund's Incomplete Adjuvant and Complete Adjuvant (Difco Laboratories, Detroit, Mich.); Merck Adjuvant 65 (Merck and Company, Inc., Rahway, N.J.); AS-2 (SmithKline Beecham); mineral salts (for example, aluminum, silica, kaolin, and carbon); aluminum salts such as aluminum hydroxide gel (alum), A1K(SO4)2, AlNa(SO4)2, A1NH4(SO4), and A1(OH)3; salts of calcium (e.g., Ca3(PO4)2), iron or zinc; an insoluble suspension of acylated tyrosine; acylated sugars; cationically or anionically derivatized polysaccharides; polynucleotides (for example, poly IC and poly AU acids); polyphosphazenes; cyanoacrylates; polymerase-(DL-lactide-co- glycoside); biodegradable microspheres; liposomes; lipid A and its derivatives; monophosphoryl lipid A; wax D from Mycobacterium tuberculosis, as well as substances found in Corynebacterium parvum, Bordetella pertussis, and members of the genus Brucella); bovine serum albumin; diphtheria toxoid; tetanus toxoid; edestin; keyhole-limpet hemocyanin; Pseudomonal Toxin A; choleragenoid; cholera toxin; pertussis toxin; viral proteins; and Quil A. Aminoalkyl glucosamine phosphate compounds can also be used (see, e.g., WO 98 / 50399, U.S. Pat. No. 6,113,918 (which issued from U.S. Ser. No. 08 / 853,826), and U.S. Ser. No. 09 / 074,720). In addition, adjuvants such as cytokines (e.g., GM-CSF or interieukin-2, -7, or -12), interferons, or tumor necrosis factor, may also be used as adjuvants.

[0075] Protein and polypeptide adjuvants may be obtained from natural or recombinant sources according to methods well known to those skilled in the art. When obtained from recombinant sources, the adjuvant may comprise a protein fragment comprising at least the immunostimulatory portion of the molecule. Other known immunostimulatory macromolecules which can be used include, but are not limited to, polysaccharides, tRNA, non-metabolizable synthetic polymers such as polyvinylamine, polymethacrylic acid, polyvinylpyrrolidone, mixed poly condensates (with relatively high molecular weight) of 4',4-diaminodiphenylmethane-3,3'-dicarboxylic acid and 4-nitro-2- aminobenzoic acid (See, Sela, M., Science 166: 1365-1374 (1969)) or glycolipids, lipids, orATTORNEY DOCKET NO. 37759.0626P1 carbohydrates.

[0132] Vaccine preparation is a well-developed art and general guidance in the preparation and formulation of vaccines is readily available from any of a variety of sources. One such example is New Trends and Developments in Vaccines, edited by Voller et al., University Park Press, Baltimore. Md.. U.S.A. 1978.

[0133] The vaccines of the present disclosure may also contain other compounds, which may be biologically active or inactive. For example, one or more immunogenic portions of other antigens may be present, either incorporated into a fusion polypeptide or as a separate compound, within the vaccine. The vaccines may generally be used for prophylactic and therapeutic purposes.

[0134] The vaccines may be formulated for any appropriate manner of administration, and thus may be administered by various methods, including for example, topical, oral, nasal, intravenous, intravaginal, epicutaneous, sublingual, intracranial, intradermal, intraperitoneal, subcutaneous, intramuscular administration, or via inhalation.

[0135] In some instances, the compositions can further comprise a pharmaceutically acceptable carrier. By “pharmaceutically acceptable” is meant a material or carrier that would be selected to minimize any degradation of the active ingredient and to minimize any adverse side effects in the subject, as would be well known to one of skill in the art. Examples of earners include dimyristoylphosphatidyl (DMPC), phosphate buffered saline or a multivesicular liposome. For example, PG:PC:Cholesterol:peptide or PC:peptide can be used as carriers in this invention. Other suitable pharmaceutically acceptable carriers and their formulations are described in Remington: The Science and Practice of Pharmacy (19th ed.) ed. A.R. Gennaro, Mack Publishing Company, Easton, PA 1995. Typically, an appropriate amount of pharmaceutically-acceptable salt is used in the formulation to render the formulation isotonic. Other examples of the pharmaceutically-acceptable carrier include, but are not limited to, saline, Ringer's solution and dextrose solution. The pF! of the solution can be from about 5 to about 8, or from about 7 to about 7.5. Further carriers include sustained release preparations such as semi-permeable matrices of solid hydrophobic polymers containing the composition, which matrices are in the form of shaped articles, e.g., films, stents (which are implanted in vessels during an angioplasty procedure), liposomes or microparticles. It will be apparent to those persons skilled in the art that certain carriers may be more preferable depending upon, for instance, the route of administration and concentration of composition being administered. These most typically would be standard carriers for administration of drugs to humans, including solutions such as sterile water, saline, and buffered solutions at phy siological pH.ATTORNEY DOCKET NO. 37759.0626P1

[0136] Pharmaceutical compositions can also include earners, thickeners, diluents, buffers, preservatives and the like, as long as the intended activity of the polypeptide, peptide, nucleic acid, vector of the invention is not compromised. Pharmaceutical compositions may also include one or more active ingredients (in addition to the composition of the invention) such as antimicrobial agents, anti-inflammatory agents, anesthetics, and the like. The pharmaceutical composition may be administered in a number of ways depending on whether local or systemic treatment is desired, and on the area to be treated.

[0137] Preparations of parenteral administration include sterile aqueous or non-aqueous solutions, suspensions, and emulsions. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Aqueous carriers include water, alcoholic / aqueous solutions, emulsions or suspensions, including saline and buffered media. Parenteral vehicles include sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, lactated Ringer’s, or fixed oils. Intravenous vehicles include fluid and nutrient replenishers, electrolyte replenishers (such as those based on Ringer’s dextrose), and the like. Preservatives and other additives may also be present such as, for example, antimicrobials, anti-oxidants, chelating agents, and inert gases and the like.

[0138] Formulations for optical administration may include ointments, lotions, creams, gels, drops, suppositories, sprays, liquids and powders. Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners and the like may be necessary or desirable.

[0139] Compositions for oral administration include powders or granules, suspensions or solutions in water or non-aqueous media, capsules, sachets, or tablets. Thickeners, flavorings, diluents, emulsifiers, dispersing aids, or binders may be desirable. Some of the compositions may potentially be administered as a pharmaceutically acceptable acid- or base- addition salt, formed by reaction with inorganic acids such as hydrochloric acid, hydrobromic acid, perchloric acid, nitric acid, thiocyanic acid, sulfuric acid, and phosphoric acid, and organic acids such as formic acid, acetic acid, propionic acid, glycolic acid, lactic acid, pyruvic acid, oxalic acid, malonic acid, succinic acid, maleic acid, and fumaric acid, or by reaction with an inorganic base such as sodium hydroxide, ammonium hydroxide, potassium hydroxide, and organic bases such as mon-, di-, trialkyl and aryl amines and substituted ethanolamines.

[0140] Thus, the compositions disclosed herein can comprise lipids such as liposomes, such as cationic liposomes (e.g., DOTMA, DOPE, DC-cholesterol) or anionic liposomes. Liposomes can further comprise proteins to facilitate targeting a particular cell, if desired. Administration of a composition comprising a peptide and a cationic liposome can be administered to the blood, to a target organ, or inhaled into the respiratory tract to target cells of the respiratory' tract. ForATTORNEY DOCKET NO. 37759.0626P1 example, a composition comprising a peptide or nucleic acid sequence described herein and a cationic liposome can be administered to a subject's lung cells. Regarding liposomes, see, e.g., Brigham et al. Am. J. Resp. Cell. Mol. Biol. 1:95 100 (1989); Feigner et al. Proc. Natl. Acad. Sci USA 84:7413 7417 (1987); U.S. Patent No. 4.897,355. Furthermore, the compound can be administered as a component of a microcapsule that can be targeted to specific cell types, such as macrophages, or where the diffusion of the compound or delivery of the compound from the microcapsule is designed for a specific rate or dosage.

[0141] In some instances, disclosed are pharmaceutical compositions comprising any of the disclosed peptides described herein, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable earner, buffer, or diluent. In various aspects, the peptide of the pharmaceutical composition is encapsulated in a delivery vehicle. In a further aspect, the delivery vehicle is a liposome, a microcapsule, or a nanoparticle. In a still further aspect, the delivery vehicle is PEG-ylated.

[0142] In the methods described herein, delivery of the compositions to cells can be via a variety of mechanisms. As defined above, disclosed herein are compositions comprising any one or more of the peptides described herein and can also include a carrier such as a pharmaceutically acceptable carrier. For example, disclosed are pharmaceutical compositions, comprising the peptides disclosed herein, and a pharmaceutically acceptable carrier. In one aspect, disclosed are pharmaceutical compositions comprising the disclosed peptides. That is, a pharmaceutical composition can be provided comprising a therapeutically effective amount of at least one disclosed peptide or at least one product of a disclosed method and a pharmaceutically acceptable carrier.

[0143] In certain aspects, the disclosed pharmaceutical compositions comprise the disclosed peptides (including pharmaceutically acceptable salt(s) thereof) as an active ingredient, a pharmaceutically acceptable carrier, and, optionally, other therapeutic ingredients or adjuvants. The instant compositions include those suitable for nasal, oral, rectal, topical, and parenteral (including subcutaneous, intramuscular, and intravenous) administration, although the most suitable route in any given case will depend on the particular host, and nature and severity of the conditions for which the active ingredient is being administered. The pharmaceutical compositions can be conveniently presented in unit dosage form and prepared by any of the methods well known in the art of pharmacy.

[0144] In practice, the peptides described herein, or pharmaceutically acceptable salts thereof, of this invention can be combined as the active ingredient in intimate admixture with a pharmaceutical carrier according to conventional pharmaceutical compounding techniques. TheATTORNEY DOCKET NO. 37759.0626P1 carrier can take a wide variety of forms depending on the form of preparation desired for administration, e.g., oral or parenteral (including intravenous). Thus, the pharmaceutical compositions of the present invention can be presented as discrete units suitable for oral administration such as capsules, cachets or tablets each containing a predetermined amount of the active ingredient. Further, the compositions can be presented as a powder, as granules, as a solution, as a suspension in an aqueous liquid, as a non-aqueous liquid, as an oil-in-water emulsion or as a water-in-oil liquid emulsion. In addition to the common dosage forms set out above, the compounds of the invention, and / or pharmaceutically acceptable salt(s) thereof, can also be administered by controlled release means and / or delivery devices. The compositions can be prepared by any of the methods of pharmacy. In general, such methods include a step of bringing into association the active ingredient with the carrier that constitutes one or more necessary' ingredients. In general, the compositions are prepared by uniformly and intimately admixing the active ingredient with liquid carriers or finely divided solid carriers or both. The product can then be conveniently shaped into the desired presentation.

[0145] By “pharmaceutically acceptable” is meant a material or carrier that would be selected to minimize any degradation of the active ingredient and to minimize any adverse side effects in the subject, as would be well known to one of skill in the art. The peptides described herein, or pharmaceutically acceptable salts thereof, can also be included in pharmaceutical compositions in combination with one or more other therapeutically active compounds.

[0146] The pharmaceutical carrier employed can be, for example, a solid, liquid, or gas. Examples of solid carriers include lactose, terra alba, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, and stearic acid. Examples of liquid carriers are sugar syrup, peanut oil. olive oil, and water. Examples of gaseous carriers include carbon dioxide and nitrogen. Other examples of carriers include dimyristoylphosphatidyl (DMPC), phosphate buffered saline or a multivesicular liposome. For example, PG:PC:Cholesterol:peptide or PC:peptide can be used as carriers in this invention. Other suitable pharmaceutically acceptable carriers and their formulations are described in Remington: The Science and Practice of Pharmacy (19th ed.) ed. A.R. Gennaro, Mack Publishing Company, Easton, PA 1995. Typically, an appropriate amount of pharmaceutically-acceptable salt is used in the formulation to render the formulation isotonic. Other examples of the pharmaceutically-acceptable carrier include, but are not limited to, saline, Ringer's solution and dextrose solution. The pH of the solution can be from about 5 to about 8, or from about 7 to about 7.5. Further carriers include sustained release preparations such as semi-permeable matrices of solid hydrophobic polymers containing the composition, which matrices are in the form of shaped articles, e.g., films, stents (which are implanted in vesselsATTORNEY DOCKET NO. 37759.0626P1 during an angioplasty procedure), liposomes or microparticles. It will be apparent to those persons skilled in the art that certain carriers may be more preferable depending upon, for instance, the route of administration and concentration of composition being administered. These most typically would be standard carriers for administration of drugs to humans, including solutions such as sterile water, saline, and buffered solutions at physiological pH.

[0147] In order to enhance the solubility and / or the stability' of the disclosed peptides in pharmaceutical compositions, it can be advantageous to employ a-, (3- or y-cyclodextrins or their derivatives, in particular hydroxyalkyl substituted cyclodextrins, e.g. 2-hydro\ypropyl-[3- cyclodextrin or sulfobutyl- -cyclodextrin. Also, co-solvents such as alcohols may improve the solubility and / or the stability of the compounds according to the invention in pharmaceutical compositions.

[0148] Pharmaceutical compositions can also include carriers, thickeners, diluents, buffers, preservatives and the like, as long as the intended activity of the polypeptide, peptide, nucleic acid, vector of the invention is not compromised. Pharmaceutical compositions may also include one or more active ingredients (in addition to the composition of the invention) such as antimicrobial agents, anti-inflammatory agents, anesthetics, and the like. The pharmaceutical composition may be administered in a number of ways depending on whether local or systemic treatment is desired, and on the area to be treated.

[0149] Because of the ease in administration, oral administration can be used, and tablets and capsules represent the most advantageous oral dosage unit forms in which case solid pharmaceutical carriers are obviously employed. In preparing the compositions for oral dosage form, any convenient pharmaceutical media can be employed. For example, water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents and the like can be used to form oral liquid preparations such as suspensions, elixirs and solutions; while carriers such as starches, sugars, microctystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrating agents, and the like can be used to form oral solid preparations such as powders, capsules and tablets. Because of their ease of administration, tablets and capsules are the preferred oral dosage units whereby solid pharmaceutical carriers are employed. Optionally, tablets can be coated by standard aqueous or nonaqueous techniques.

[0150] Compositions for oral administration include powders or granules, suspensions or solutions in water or non-aqueous media, capsules, sachets, or tablets. Thickeners, flavorings, diluents, emulsifiers, dispersing aids, or binders may be desirable. Some of the compositions may potentially be administered as a pharmaceutically acceptable acid- or base- addition salt, formed by reaction with inorganic acids such as hydrochloric acid, hydrobromic acid, perchloricATTORNEY DOCKET NO. 37759.0626P1 acid, nitric acid, thiocyanic acid, sulfuric acid, and phosphoric acid, and organic acids such as formic acid, acetic acid, propionic acid, glycolic acid, lactic acid, pyruvic acid, oxalic acid, malonic acid, succinic acid, maleic acid, and fumaric acid, or by reaction with an inorganic base such as sodium hydroxide, ammonium hydroxide, potassium hydroxide, and organic bases such as mon-, di-, trialkyl and aryl amines and substituted ethanolamines.

[0151] A tablet containing the compositions of the present invention can be prepared by compression or molding, optionally with one or more accessory' ingredients or adjuvants. Compressed tablets can be prepared by compressing, in a suitable machine, the active ingredient in a free-flowing form such as powder or granules, optionally mixed with a binder, lubricant, inert diluent, surface active or dispersing agent. Molded tablets can be made by molding in a suitable machine, a mixture of the powdered compound moistened with an inert liquid diluent.

[0152] The pharmaceutical compositions of the present invention comprise a disclosed peptide (or pharmaceutically acceptable salts thereof) as an active ingredient, a pharmaceutically acceptable carrier, and optionally one or more additional therapeutic agents or adjuvants. The instant compositions include compositions suitable for oral, rectal, topical, and parenteral (including subcutaneous, intramuscular, and intravenous) administration, although the most suitable route in any given case will depend on the particular host, and nature and severity of the conditions for which the active ingredient is being administered. The pharmaceutical compositions can be conveniently presented in unit dosage form and prepared by any of the methods well known in the art of pharmacy.

[0153] Pharmaceutical compositions of the present invention suitable for parenteral administration can be prepared as solutions or suspensions of the active compounds in water. A suitable surfactant can be included such as, for example, hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof in oils. Further, a preservative can be included to prevent the detrimental grow th of microorganisms.

[0154] Pharmaceutical compositions of the present invention suitable for injectable use include sterile aqueous solutions or dispersions. Furthermore, the compositions can be in the form of sterile powders for the extemporaneous preparation of such sterile injectable solutions or dispersions. Typically, the final injectable form should be sterile and should be effectively fluid for easy syringability . The pharmaceutical compositions should be stable under the conditions of manufacture and storage; thus, preferably should be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (e.g., glycerol, propylene glycol and liquid polyethylene glycol), vegetable oils, and suitable mixtures thereof.ATTORNEY DOCKET NO. 37759.0626P1

[0155] Injectable solutions, for example, can be prepared in which the carrier comprises saline solution, glucose solution or a mixture of saline and glucose solution. Injectable suspensions may also be prepared in which case appropriate liquid carriers, suspending agents and the like may be employed. Also included are solid form preparations that are intended to be converted, shortly before use, to liquid form preparations.

[0156] Preparations of parenteral administration include sterile aqueous or non-aqueous solutions, suspensions, and emulsions. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Aqueous carriers include water, alcoholic / aqueous solutions, emulsions or suspensions, including saline and buffered media. Parenteral vehicles include sodium chloride solution, Ringer’s dextrose, dextrose and sodium chloride, lactated Ringer’s, or fixed oils. Intravenous vehicles include fluid and nutrient replenishers, electrolyte replenishers (such as those based on Ringer's dextrose), and the like. Preservatives and other additives may also be present such as. for example, antimicrobials, anti-oxidants, chelating agents, and inert gases and the like.

[0157] Pharmaceutical compositions of the present invention can be in a form suitable for topical use such as, for example, an aerosol, cream, ointment, lotion, dusting powder, mouth washes, gargles, and the like. Further, the compositions can be in a form suitable for use in transdermal devices. These formulations can be prepared, utilizing a compound of the invention, or pharmaceutically acceptable salts thereof, via conventional processing methods. As an example, a cream or ointment is prepared by mixing hydrophilic material and water, together with about 5 wt% to about 10 wt% of the compound, to produce a cream or ointment having a desired consistency.

[0158] In the compositions suitable for percutaneous administration, the carrier optionally comprises a penetration enhancing agent and / or a suitable wetting agent, optionally combined with suitable additives of any nature in minor proportions, which additives do not introduce a significant deleterious effect on the skin. Said additives may facilitate the administration to the skin and / or may be helpful for preparing the desired compositions. These compositions may be administered in various ways, e g., as a transdermal patch, as a spot on, as an ointment.

[0159] Pharmaceutical compositions of this invention can be in a form suitable for rectal administration wherein the carrier is a solid. It is preferable that the mixture forms unit dose suppositories. Suitable carriers include cocoa butter and other materials commonly used in the art. The suppositories can be conveniently formed by first admixing the composition with the softened or melted carrier(s) followed by chilling and shaping in molds.

[0160] Formulations for optical administration may include ointments, lotions, creams, gels,ATTORNEY DOCKET NO. 37759.0626P1 drops, suppositories, sprays, liquids and powders. Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners and the like may be desirable.

[0161] In addition to the aforementioned carrier ingredients, the pharmaceutical formulations described above can include, as appropriate, one or more additional carrier ingredients such as diluents, buffers, flavoring agents, binders, surface-active agents, thickeners, lubricants, preservatives (including anti-oxidants) and the like. Furthermore, other adjuvants can be included to render the formulation isotonic with the blood of the intended recipient. Compositions containing a disclosed peptide, and / or pharmaceutically acceptable salts thereof, can also be prepared in powder or liquid concentrate form.

[0162] The exact dosage and frequency of administration depends on the particular disclosed peptide, a product of a disclosed method of making, a pharmaceutically acceptable salt, solvate, or polymorph thereof, a hydrate thereof, a solvate thereof, a polymorph thereof, or a stereochemically isomeric form thereof; the particular condition being treated and the severity of the condition being treated; various factors specific to the medical history of the subject to whom the dosage is administered such as the age; weight, sex, extent of disorder and general physical condition of the particular subject, as well as other medication the individual may be taking; as is well known to those skilled in the art. Furthermore, it is evident that said effective daily amount may be lowered or increased depending on the response of the treated subject and / or depending on the evaluation of the physician prescribing the compositions.

[0163] Depending on the mode of administration, the pharmaceutical composition will comprise from 0.05 to 99 % by weight, preferably from 0.1 to 70 % by weight, more preferably from 0. 1 to 50 % by weight of the active ingredient, and, from 1 to 99.95 % by weight, preferably from 30 to 99.9 % by weight, more preferably from 50 to 99.9 % by weight of a pharmaceutically acceptable carrier, all percentages being based on the total weight of the composition.G. Methods

[0164] Disclosed are methods of use for the disclosed peptides, nucleic acids, vectors, compositions, or VLPs.1. Methods of neutralizing the cytotoxicity of LukGH

[0165] In some aspects, the LukGH dimer of Staphylococcus aureus is toxic to cells and leads to cytotoxicity. Thus, targeting LukGH can result in a decrease or neutralization of the cytotoxicity. LukGH can be targeted by targeting the dimer or one of the individual subunits, LukG or LukH. Described herein are methods of targeting LukGH by eliciting antibodies to LukH.ATTORNEY DOCKET NO. 37759.0626P1

[0166] Disclosed are methods of neutralizing the cytotoxicity of leukotoxin LukGH in a subject comprising administering to the subject one or more of the disclosed compositions, peptides, nucleic acids, or VLPs, wherein the subject generates neutralizing antibodies (nAbs) against LukH. In some aspects, the subject has been infected with or exposed to Staphylococcus aureus. In some aspects, the subject is at risk for being infected with or exposed to Staphylococcus aureus.

[0167] Disclosed are methods of neutralizing the cytotoxicity of leukotoxin LukGH in a subject infected with or at risk of being exposed to Staphylococcus aureus comprising administering to the subject a composition comprising a peptide comprising or consisting essentially of the amino acid sequence STAPD (SEQ ID NO: 1), STAPDD (SEQ ID NO:2), TNKDKNSTAPDDIGKNGK (SEQ ID NO:3), KDKNSTAPDDIGKNGKI (SEQ ID NO:4), STAPDDIGKNGKITKRT (SEQ ID NO:5), KEKRNVTNKDKNSTAPD (SEQ ID NO: 10), KEKR (SEQ ID NO;6), KDKR (SEQ ID NO: 7), EHVDKSQQKEKRNVTNK (SEQ ID NO: 8), or KSQQKEKRNTNKDKNS (SEQ ID NO:9) or a peptide equivalent thereof, wherein the peptide equivalent thereof is a peptide comprising or consisting essentially of an amino acid sequence at least 90% identical to STAPD (SEQ ID NO: 1), STAPDD (SEQ ID NO:2), TNKDKNSTAPDDIGKNGK (SEQ ID NO:3), KDKNSTAPDDIGKNGKI (SEQ ID NO:4), STAPDDIGKNGKITKRT (SEQ ID NO:5). KEKRNVTNKDKNSTAPD (SEQ ID NO: 10), KEKR (SEQ ID NOV), KDKR (SEQ ID NO:7), EHVDKSQQKEKRNVTNK (SEQ ID NO: 8), or KSQQKEKRNTNKDKNS (SEQ ID NOV), wherein the subject generates neutralizing antibodies (nAbs) against LukH. Thus, in some aspects, the composition is referred to as a vaccine. In some aspects, neutralizing antibodies can prevent infection with Staphylococcus aureus or treat a subject infected with Staphylococcus aureus.

[0168] In some aspects, administering a composition comprising a peptide comprising or consisting essentially of the amino acid sequence STAPD (SEQ ID NO: 1), STAPDD (SEQ ID NO:2), TNKDKNSTAPDDIGKNGK (SEQ ID NO:3), KDKNSTAPDDIGKNGKI (SEQ ID NO:4). STAPDDIGKNGKITKRT (SEQ ID NO:5). KEKRNVTNKDKNSTAPD (SEQ ID NO: 10), KEKR (SEQ ID NOV), KDKR (SEQ ID NOV), EHVDKSQQKEKRNVTNK (SEQ ID NO: 8), or KSQQKEKRNTNKDKNS (SEQ ID NOV) or a peptide equivalent thereof, wherein the peptide equivalent thereof is a peptide comprising or consisting essentially of an amino acid sequence at least 90% identical to STAPD (SEQ ID NO: 1). STAPDD (SEQ ID NO: 2). TNKDKNSTAPDDIGKNGK (SEQ ID NO:3, KDKNSTAPDDIGKNGKI (SEQ ID NO:4), STAPDDIGKNGKITKRT (SEQ ID NOV), KEKRNVTNKDKNSTAPD (SEQ ID NO: 10), KEKR (SEQ ID NOV), KDKR (SEQ ID NO: 7), EHVDKSQQKEKRNVTNK (SEQ ID NO: 8),ATTORNEY DOCKET NO. 37759.0626P1 or KSQQKEKRNTNKDKNS (SEQ ID NO:9), wherein the subject generates nAbs against LukH, results in targeting the LukH subunit of the LukGH toxin and ultimately neutralizes the entire toxin.

[0169] Disclosed are methods of neutralizing the cytotoxicity of leukotoxin LukGH in a subject infected with or at risk of being exposed to Staphylococcus aureus comprising administering to the subject two or more compositions, wherein each composition comprises a peptide comprising or consisting essentially of the amino acid sequence STAPD (SEQ ID NO:1), STAPDD (SEQ ID NO:2), TNKDKNSTAPDDIGKNGK (SEQ ID NO:3), KDKNSTAPDDIGKNGKI (SEQ ID NO:4), STAPDDIGKNGKITKRT (SEQ ID NO:5), KEKRNVTNKDKNSTAPD (SEQ ID NO: 10), KEKR (SEQ ID NO: 6), KDKR (SEQ ID NO: 7), EHVDKSQQKEKRNVTNK (SEQ ID NO:8), or KSQQKEKRNTNKDKNS (SEQ ID NO:9) or a peptide equivalent thereof, wherein the peptide equivalent thereof is a peptide comprising or consisting essentially of an amino acid sequence at least 90% identical to STAPD (SEQ ID NO:1), STAPDD (SEQ ID NO:2), TNKDKNSTAPDDIGKNGK (SEQ ID NOV), KDKNSTAPDDIGKNGKI (SEQ ID NO:4), STAPDDIGKNGKITKRT (SEQ ID NO:5), KEKRNVTNKDKNSTAPD (SEQ ID NO: 10), KEKR (SEQ ID NO: 6), KDKR (SEQ ID NO: 7), EHVDKSQQKEKRNVTNK (SEQ ID NO:8), or KSQQKEKRNTNKDKNS (SEQ ID NOV), and wherein each of the two or more compositions is different. For example, the method can comprise administering to the subject a composition, such as a VLP, comprising a peptide comprising or consisting essentially of the amino acid sequence STAPD (SEQ ID NO:1) and a composition comprising a peptide comprising or consisting essentially of the amino acid sequence KERK (SEQ ID NO: 6).

[0170] In some aspects, the composition comprising a peptide comprising or consisting essentially of the amino acid sequence STAPD (SEQ ID NO: 1), STAPDD (SEQ ID NOV), TNKDKNSTAPDDIGKNGK (SEQ ID NO:3), KDKNSTAPDDIGKNGKI (SEQ ID NO:4), STAPDDIGKNGKITKRT (SEQ ID NO:5), KEKRNVTNKDKNSTAPD (SEQ ID NO: 10), KEKR (SEQ ID NOV), KDKR (SEQ ID NOV), EHVDKSQQKEKRNVTNK (SEQ ID NO: 8), or KSQQKEKRNTNKDKNS (SEQ ID NOV), or a peptide equivalent thereof is a VLP. Thus, disclosed are methods of neutralizing the cytotoxicity of leukotoxin LukGH in a subject infected with or at risk of being exposed to Staphylococcus aureus comprising administering to the subject a VLP displaying one or more of peptides comprising or consisting essentially of the ammo acid sequence STAPD (SEQ ID NO:1), STAPDD (SEQ ID NOV), TNKDKNSTAPDDIGKNGK (SEQ ID NO:3), KDKNSTAPDDIGKNGKI (SEQ ID NO:4), STAPDDIGKNGKITKRT (SEQ ID NOV), KEKRNVTNKDKNSTAPD (SEQ ID NO: 10),ATTORNEY DOCKET NO. 37759.0626P1KEKR (SEQ ID N0:6), KDKR (SEQ ID N0:7), EHVDKSQQKEKRNVTNK (SEQ ID NO: 8), or KSQQI<EI<RNTNI<DI<NS (SEQ ID NOV).2. Methods of Treating

[0171] Disclosed are methods of treating a subject infected with Staphylococcus aureus comprising administering to the subject one or more of the disclosed immunogenic compositions, peptides, nucleic acids, or VLPs.

[0172] Disclosed are methods of treating a subject infected with Staphylococcus aureus comprising administering to the subject a composition comprising a peptide comprising or consisting essentially of the amino acid sequence STAPD (SEQ ID NO:1), STAPDD (SEQ ID NO: 2), TNKDKNSTAPDDIGKNGK (SEQ ID NO: 3), KDKNSTAPDDIGKNGKI (SEQ ID NO:4), STAPDDIGKNGKITKRT (SEQ ID NO:5), KEKRNVTNKDKNSTAPD (SEQ ID NO: 10), KEKR (SEQ ID NO:6), KDKR (SEQ ID NOV). EHVDKSQQKEKRNVTNK (SEQ ID NO:8), or KSQQKEKRNTNKDKNS (SEQ ID NO:9), or a peptide equivalent thereof, wherein the peptide equivalent thereof is a peptide comprising or consisting essentially of an amino acid sequence at least 90% identical to STAPD (SEQ ID NO:1), STAPDD (SEQ ID NOV), TNKDKNSTAPDDIGKNGK (SEQ ID NO:3), KDKNSTAPDDIGKNGKI (SEQ ID NO:4), STAPDDIGKNGKITKRT (SEQ ID NO:5). KEKRNVTNKDKNSTAPD (SEQ ID NO: 10), KEKR (SEQ ID NO:6), KDKR (SEQ ID NOV), EHVDKSQQKEKRNVTNK (SEQ ID NO: 8), or KSQQKEKRNTNKDKNS (SEQ ID NOV).

[0173] In some aspects, administering a composition comprising a peptide comprising or consisting essentially of the amino acid sequence STAPD (SEQ ID NO:1), STAPDD (SEQ ID NOV), TNKDKNSTAPDDIGKNGK (SEQ ID NOV), KDKNSTAPDDIGKNGKI (SEQ ID NO:4), STAPDDIGKNGKITKRT (SEQ ID NO:5), KEKRNVTNKDKNSTAPD (SEQ ID NO: 10), KEKR (SEQ ID NO:6), KDKR (SEQ ID NOV), EHVDKSQQKEKRNVTNK (SEQ ID NO: 8), or KSQQKEKRNTNKDKNS (SEQ ID NOV), or a peptide equivalent thereof, wherein the peptide equivalent thereof is a peptide comprising or consisting essentially of an amino acid sequence at least 90% identical to STAPD (SEQ ID NO:1), STAPDD (SEQ ID NOV), TNKDKNSTAPDDIGKNGK (SEQ ID NO:3), KDKNSTAPDDIGKNGKI (SEQ ID NOV), STAPDDIGKNGKITKRT (SEQ ID NO:5), KEKRNVTNKDKNSTAPD (SEQ ID NO: 10), KEKR (SEQ ID NO:6), KDKR (SEQ ID NOV), EHVDKSQQKEKRNVTNK (SEQ ID NO: 8), or KSQQKEKRNTNKDKNS (SEQ ID NOV), results in the subject generating neutralizing antibodies (nAbs) against LukH. nAb to LukH target the LukH subunit of the LukGH toxin and ultimately neutralizes the entire toxin.ATTORNEY DOCKET NO. 37759.0626P1

[0174] Disclosed are methods of treating a subject infected with Staphylococcus aureus comprising administering to the subject two or more compositions, wherein each composition comprises a peptide comprising or consisting essentially of the amino acid sequence STAPD (SEQ ID NO:1), STAPDD (SEQ ID NO:2), TNKDKNSTAPDDIGKNGK (SEQ ID NOT), KDKNSTAPDDIGKNGKI (SEQ ID NO:4), STAPDDIGKNGKITKRT (SEQ ID NOV), KEKRNVTNKDKNSTAPD (SEQ ID NO: 10), KEKR (SEQ ID NO: 6), KDKR (SEQ ID NO: 7), EHVDKSQQKEKRNVTNK (SEQ ID NO: 8), or KSQQKEKRNTNKDKNS (SEQ ID NO: 9), or a peptide equivalent thereof, wherein the peptide equivalent thereof is a peptide comprising or consisting essentially of an amino acid sequence at least 90% identical to STAPD (SEQ ID NOT), STAPDD (SEQ ID NOT), TNKDKNSTAPDDIGKNGK (SEQ ID NOT), KDKNSTAPDDIGKNGKI (SEQ ID NO:4), STAPDDIGKNGKITKRT (SEQ ID NO:5), KEKRNVTNKDKNSTAPD (SEQ ID NOTO), KEKR (SEQ ID NO: 6), KDKR (SEQ ID NOT), EHVDKSQQKEKRNVTNK (SEQ ID NO: 8), or KSQQKEKRNTNKDKNS (SEQ ID NOV), and wherein each of the two or more compositions is different. For example, the method can comprise administering to the subject a composition, such as a VLP, comprising a peptide comprising or consisting essentially of the amino acid sequence STAPD (SEQ ID NO:1) and a composition comprising a peptide comprising or consisting essentially of the amino acid sequence KEKR (SEQ ID NO: 6).

[0175] In some aspects, the composition comprising a peptide comprising or consisting essentially of the amino acid sequence STAPD (SEQ ID NO: 1), STAPDD (SEQ ID NOT), TNKDKNSTAPDDIGKNGK (SEQ ID NOT), KDKNSTAPDDIGKNGKI (SEQ ID NO:4), STAPDDIGKNGKITKRT (SEQ ID NO:5). KEKRNVTNKDKNSTAPD (SEQ ID NOTO), KEKR (SEQ ID NO:6), KDKR (SEQ ID NOT), EHVDKSQQKEKRNVTNK (SEQ ID NO: 8), or KSQQKEKRNTNKDKNS (SEQ ID NOV), or a peptide equivalent thereof is a VLP. Thus, disclosed are methods of treating a subj ect infected with Staphylococcus aureus comprising administering to the subject a VLP displaying one or more of peptides comprising or consisting essentially of the amino acid sequence STAPD (SEQ ID NO: 1), STAPDD (SEQ ID NOT), TNKDKNSTAPDDIGKNGK (SEQ ID NOT), KDKNSTAPDDIGKNGKI (SEQ ID NO:4), STAPDDIGKNGKITKRT (SEQ ID NOV), KEKRNVTNKDKNSTAPD (SEQ ID NOTO), KEKR (SEQ ID NO:6), KDKR (SEQ ID NOT), EHVDKSQQKEKRNVTNK (SEQ ID NO: 8), or KSQQKEKRNTNKDKNS (SEQ ID NOV).

[0176] In some aspects, the methods of treating can further comprise administering one or more antibiotics. In some aspects, the antibiotic can be, but is not limited to, tetracycline, doxicycline, minocycline, trimethoprim-sulfamethoxazole, rifampin, clindamycin, vancomycin,ATTORNEY DOCKET NO. 37759.0626P1 linezolicL daptomycin, tigecycline, telavancin, dalbavancin, oritavancin, ceftobiprole, mupirocin or iclaprim.

[0177] In some aspects, the methods of treating further comprise administering one or more antibodies that are specific for any Staphylococcus aureus epitope. In some aspects, the Staphylococcus aureus epitope is an alpha toxin epitope. In some aspects, the alpha toxin epitope is GNVTGDDTGKIGGLIG (SEQ ID NO: 17).

[0178] In some aspects, the methods of treating further comprise administering one or more VLPs that display a Staphylococcus aureus epitope. In some aspects, the Staphylococcus aureus epitope is an alpha toxin epitope. In some aspects, the alpha toxin epitope is GNVTGDDTGKIGGLIG (SEQ ID NO: 17). Thus, in some aspects, the methods of treating further comprise administering one or more VLPs that display the alpha toxin epitope GNVTGDDTGKIGGLIG (SEQ ID NO: 17). Thus, in some aspects, the methods comprise administering one or more of the VLPs displaying an epitope from a LukH peptide and one or more of the VLPs displaying an epitope from alpha toxin. In some aspects, the disclosed method of treating can include the combination treatment of a VLP comprising an amino acid sequence of STAPD (SEQ ID NOT), STAPDD (SEQ ID NOT), TNKDKNSTAPDDIGKNGK (SEQ ID NOT), KDKNSTAPDDIGKNGKI (SEQ ID NO:4), STAPDDIGKNGKITKRT (SEQ ID NOT), KEKRNVTNKDKNSTAPD (SEQ ID NO: 10), KEKR (SEQ ID NO:6). KDKR (SEQ ID NOT), EHVDKSQQKEKRNVTNK (SEQ ID NO: 8), or KSQQKEKRNTNKDKNS (SEQ ID NOV), or an equivalent thereof, wherein the equivalent thereof is a protein or peptide comprising or consisting essentially of an amino acid sequence at least 90% identical to STAPD (SEQ ID NOT), STAPDD (SEQ ID NO:2), TNKDKNSTAPDDIGKNGK (SEQ ID NOT), KDKNSTAPDDIGKNGKI (SEQ ID NO:4), STAPDDIGKNGKITKRT (SEQ ID NOT), KEKRNVTNKDKNSTAPD (SEQ ID NO: 10), KEKR (SEQ ID NO: 6), KDKR (SEQ ID NOT), EHVDKSQQKEKRNVTNK (SEQ ID NOT), or KSQQKEKRNTNKDKNS (SEQ ID NOV) and a VLP comprising the alpha toxin epitope GNVTGDDTGKIGGLIG (SEQ ID NOT 7).

[0179] In some aspects, the methods of treating further comprise administering one or more VLPs comprising the alpha toxin epitope is GNVTGDDTGKIGGLIG (SEQ ID NO: 17). In some aspects, like the disclosed VLPs comprising STAPD (SEQ ID NOT), STAPDD (SEQ ID NO: 2), TNKDKNSTAPDDIGKNGK (SEQ ID NOT), KDKNSTAPDDIGKNGKI (SEQ ID NO:4), STAPDDIGKNGKITKRT (SEQ ID NO:5). KEKRNVTNKDKNSTAPD (SEQ ID NOTO), KEKR (SEQ ID NO:6), KDKR (SEQ ID NOT), EHVDKSQQKEKRNVTNK (SEQ ID NOT), or KSQQKEKRNTNKDKNS (SEQ ID NOV), an alternative VLP can be used that comprises or expresses the alpha toxin epitope is GNVTGDDTGKIGGLIG (SEQ ID NO: 17).ATTORNEY DOCKET NO. 37759.0626P1Thus, in some aspects, the VLP comprising or expressing the alpha toxin epitope is GNVTGDDTGKIGGLIG (SEQ ID NO: 17), can elicit an immune response to the alpha toxin epitope. This VLP can act together with the composition comprising an immunogen comprising / consisting essentially of the STAPD (SEQ ID NO:1) epitope TNKDKNSTAPDDIGKNGK (SEQ ID NOV), KDKNSTAPDDIGKNGKI (SEQ ID NOV). STAPDDIGKNGKITKRT (SEQ ID NOV), STAPD (SEQ ID NO:1), STAPDD (SEQ ID NOV), or an immunogenic equivalent thereof, wherein the immunogenic equivalent thereof is a peptide comprising / consisting essentially of an amino acid sequence at least 90% identical to TNKDKNSTAPDDIGKNGK (SEQ ID NOV), KDKNSTAPDDIGKNGKI (SEQ ID NOV). STAPDDIGKNGKITKRT (SEQ ID NO:5), STAPD (SEQ ID NO: 1), or STAPDD (SEQ ID NO:2) to treat a subject. This VLP can act together with the composition comprising an immunogen comprising / consisting essentially of the KEKR (SEQ ID NO:6) epitope KEKRNVTNKDKNSTAPD (SEQ ID NO: 10), EHVDKSQQKEKRNVTNK (SEQ ID NO:8), KSQQKEKRNTNKDKNS (SEQ ID NOV), KEKR (SEQ ID NO:6), KDKR (SEQ ID NOV), or an immunogenic equivalent thereof, w herein the immunogenic equivalent thereof is a peptide comprising / consisting essentially of an amino acid sequence at least 90% identical to KEKRNVTNKDKNSTAPD (SEQ ID NO: 10), EHVDKSQQKEKRNVTNK (SEQ ID NO:8), KSQQKEKRNTNKDKNS (SEQ ID NOV), KEKR (SEQ ID NO:6), KDKR (SEQ ID NOV) to treat a subj ect.

[0180] In some aspects, the disclosed method of treating can include the combination treatment of a VLP comprising an amino acid sequence of STAPD (SEQ ID NO:1), STAPDD (SEQ ID NOV), TNKDKNSTAPDDIGKNGK (SEQ ID NOV), KDKNSTAPDDIGKNGKI (SEQ ID NOV), STAPDDIGKNGKITKRT (SEQ ID NOV), KEKRNVTNKDKNSTAPD (SEQ ID NOVO), KEKR (SEQ ID NO:6), KDKR (SEQ ID NOV), EHVDKSQQKEKRNVTNK (SEQ ID NOV), or KSQQKEKRNTNKDKNS (SEQ ID NOV), or an equivalent thereof, wherein the equivalent thereof is a protein or peptide comprising or consisting essentially of an amino acid sequence at least 90% identical to STAPD (SEQ ID NOV). STAPDD (SEQ ID NOV). TNKDKNSTAPDDIGKNGK (SEQ ID NOV), KDKNSTAPDDIGKNGKI (SEQ ID NOV), STAPDDIGKNGKITKRT (SEQ ID NO:5), KEKRNVTNKDKNSTAPD (SEQ ID NOVO), KEKR (SEQ ID NO:6), KDKR (SEQ ID NOV), EHVDKSQQKEKRNVTNK (SEQ ID NO: 8), or KSQQKEKRNTNKDKNS (SEQ ID NOV) and a VLP comprising the alpha toxin epitope GNVTGDDTGKIGGLIG (SEQ ID NOV 7).3. Methods of Preventing

[0181] Disclosed are methods of preventing a Staphylococcus aureus infection in a subjectATTORNEY DOCKET NO. 37759.0626P1 comprising administering to the subject one or more of the disclosed immunogenic compositions, peptides, nucleic acids, or VLPs. In some aspects, the subject is at risk of Staphylococcus aureus infection or exposure to Staphylococcus aureus.

[0182] Disclosed are methods of preventing a Staphylococcus aureus infection in a subject comprising administering to the subject a composition comprising a peptide comprising or consisting essentially of the amino acid sequence STAPD (SEQ ID NO: 1), STAPDD (SEQ ID NO: 2), TNKDKNSTAPDDIGKNGK (SEQ ID NO: 3), KDKNSTAPDDIGKNGKI (SEQ ID NO:4), STAPDDIGKNGKITKRT (SEQ ID NO:5), KEKRNVTNKDKNSTAPD (SEQ ID NO: 10), KEKR (SEQ ID NO:6), KDKR (SEQ ID NOV). EHVDKSQQKEKRNVTNK (SEQ ID NO: 8), or KSQQKEKRNTNKDKNS (SEQ ID NO:9), or a peptide equivalent thereof, wherein the peptide equivalent thereof is a peptide comprising or consisting essentially of an amino acid sequence at least 90% identical to STAPD (SEQ ID NO: 1), STAPDD (SEQ ID NOV), TNKDKNSTAPDDIGKNGK (SEQ ID NO:3), KDKNSTAPDDIGKNGKI (SEQ ID NO:4). STAPDDIGKNGKITKRT (SEQ ID NO:5), KEKRNVTNKDKNSTAPD (SEQ ID NO: 10), KEKR (SEQ ID NO:6), KDKR (SEQ ID NOV), EHVDKSQQKEKRNVTNK (SEQ ID NO: 8), or KSQQI<EKRNTNI<DI<NS (SEQ ID NOV).

[0183] In some aspects, administering a composition comprising a peptide comprising or consisting essentially of the amino acid sequence STAPD (SEQ ID NO: 1), STAPDD (SEQ ID NOV), TNKDKNSTAPDDIGKNGK (SEQ ID NOV), KDKNSTAPDDIGKNGKI (SEQ ID NO:4), STAPDDIGKNGKITKRT (SEQ ID NO:5), KEKRNVTNKDKNSTAPD (SEQ ID NO: 10), KEKR (SEQ ID NO:6), KDKR (SEQ ID NOV), EHVDKSQQKEKRNVTNK (SEQ ID NO: 8), or KSQQKEKRNTNKDKNS (SEQ ID NOV), or a peptide equivalent thereof, wherein the peptide equivalent thereof is a peptide comprising or consisting essentially of an amino acid sequence at least 90% identical to STAPD (SEQ ID NO: 1), STAPDD (SEQ ID NOV), TNKDKNSTAPDDIGKNGK (SEQ ID NO:3), KDKNSTAPDDIGKNGKI (SEQ ID NO:4), STAPDDIGKNGKITKRT (SEQ ID NO:5), KEKRNVTNKDKNSTAPD (SEQ ID NO: 10), KEKR (SEQ ID NO:6), KDKR (SEQ ID NOV), EHVDKSQQKEKRNVTNK (SEQ ID NO: 8), or KSQQKEKRNTNKDKNS (SEQ ID NOV), results in the subject generating neutralizing antibodies (nAbs) against LukH. nAb to LukH target the LukH subunit of the LukGH toxin. In some aspects, the nAbs can neutralizes the entire toxin LukGH toxin if the subject is exposed to Staphylococcus aureus.

[0184] Disclosed are methods of preventing a Staphylococcus aureus infection in a subject comprising administering to the subject two or more compositions, wherein each composition comprises a peptide comprising or consisting essentially of the amino acid sequence STAPDATTORNEY DOCKET NO. 37759.0626P1(SEQ ID NO:1), STAPDD (SEQ ID NOV), TNKDKNSTAPDDIGKNGK (SEQ ID N0:3), KDKNSTAPDDIGKNGKI (SEQ ID N0:4), STAPDDIGKNGKITKRT (SEQ ID N0:5), KEKRNVTNKDKNSTAPD (SEQ ID NO: 10), KEKR (SEQ ID NO: 6), KDKR (SEQ ID NO: 7), EHVDKSQQKEKRNVTNK (SEQ ID NO: 8), or KSQQKEKRNTNKDKNS (SEQ ID NO: 9), or a peptide equivalent thereof, wherein the peptide equivalent thereof is a peptide comprising or consisting essentially of an amino acid sequence at least 90% identical to STAPD (SEQ ID NO:1), STAPDD (SEQ ID NOV), TNKDKNSTAPDDIGKNGK (SEQ ID NOV), KDKNSTAPDDIGKNGKI (SEQ ID NO:4), STAPDDIGKNGKITKRT (SEQ ID NOV), KEKRNVTNKDKNSTAPD (SEQ ID NO: 10), KEKR (SEQ ID NO:6), KDKR (SEQ ID NOV). EHVDKSQQKEKRNVTNK (SEQ ID NO: 8), or KSQQKEKRNTNKDKNS (SEQ ID NOV), and wherein each of the two or more compositions is different. For example, the method can comprise administering to the subject a composition, such as a VLP, comprising a peptide comprising or consisting essentially of the amino acid sequence STAPD and a composition comprising a peptide comprising or consisting essentially of the amino acid sequence KEKR.

[0185] In some aspects, the composition comprising a peptide comprising or consisting essentially of the amino acid sequence STAPD (SEQ ID NO: 1), STAPDD (SEQ ID NOV), TNKDKNSTAPDDIGKNGK (SEQ ID NOV), KDKNSTAPDDIGKNGKI (SEQ ID NO:4), STAPDDIGKNGKITKRT (SEQ ID NOV), KEKRNVTNKDKNSTAPD (SEQ ID NO: 10), KEKR (SEQ ID NO:6), KDKR (SEQ ID NOV), EHVDKSQQKEKRNVTNK (SEQ ID NO: 8), or KSQQKEKRNTNKDKNS (SEQ ID NOV), or a peptide equivalent thereof, is a VLP assembled particle comprising one or more of the disclosed VLP monomers. For example, the composition comprising a peptide comprising or consisting essentially of the amino acid sequence STAPD (SEQ ID NO:1), STAPDD (SEQ ID NOV), TNKDKNSTAPDDIGKNGK (SEQ ID NOV), KDKNSTAPDDIGKNGKI (SEQ ID NO:4), STAPDDIGKNGKITKRT (SEQ ID NOV), KEKRNVTNKDKNSTAPD (SEQ ID NOTO), KEKR (SEQ ID NOV), KDKR (SEQ ID NOV), EHVDKSQQKEKRNVTNK (SEQ ID NO: 8), or KSQQKEKRNTNKDKNS (SEQ ID NOV), or a peptide equivalent thereof, is a VLP assembled particle comprising VLP monomers comprising one or more of epitopes STAPD (SEQ ID NO:1), STAPDD (SEQ ID NOV), TNKDKNSTAPDDIGKNGK (SEQ ID NO: 3), KDKNSTAPDDIGKNGKI (SEQ ID NO:4), STAPDDIGKNGKITKRT (SEQ ID NO:5), KEKRNVTNKDKNSTAPD (SEQ ID NO: 10), KEKR (SEQ ID NOV), KDKR (SEQ ID NOV). EHVDKSQQKEKRNVTNK (SEQ ID NO: 8), or KSQQKEKRNTNKDKNS (SEQ ID NOV).

[0186] In some aspects, preventing a Staphylococcus aureus infection involves an initial infection by the bacteria that is then prevented from further infecting or replicating within theATTORNEY DOCKET NO. 37759.0626P1 subject and thus prevents any health complications in the subject. In some aspects, preventing a Staphylococcus aureus infection can mean clearing the initial infection from the subject within 12 to 48 hours. In some aspects, preventing a Staphylococcus aureus infection can mean no health complications are observed due to limited infection ability of the bacteria. Thus, in some aspects, a subject exposed to Staphylococcus aureus may get an initial infection of the bacteria but the bacteria is cleared from the subject within 12 to 48 hours. In some aspects, a subject exposed to Staphylococcus aureus may get an initial infection of the bacteria but no health complications are seen.

[0187] In some aspects, the methods of preventing further comprise administering one or more antibodies that are specific for any Staphylococcus aureus epitope. In some aspects, the Staphylococcus aureus epitope is an alpha toxin epitope. In some aspects, the alpha toxin epitope is GNVTGDDTGKIGGLIG (SEQ ID NO: 17).

[0188] In some aspects, the methods of preventing further comprise administering one or more VLPs comprising the alpha toxin epitope is GNVTGDDTGKIGGLIG (SEQ ID NO: 17). In some aspects, like the disclosed VLPs comprising STAPD (SEQ ID NO: 1), STAPDD (SEQ ID NO:2), TNKDKNSTAPDDIGKNGK (SEQ ID NOT), KDKNSTAPDDIGKNGKI (SEQ ID NO:4), STAPDDIGKNGKITKRT (SEQ ID NO:5), KEKRNVTNKDKNSTAPD (SEQ ID NO: 10), KEKR (SEQ ID NO:6), KDKR (SEQ ID NOT). EHVDKSQQKEKRNVTNK (SEQ ID NO: 8), or KSQQKEKRNTNKDKNS (SEQ ID NOV), an alternative VLP can be used that comprises or expresses the alpha toxin epitope GNVTGDDTGKIGGLIG (SEQ ID NO: 17). Thus, in some aspects, the VLP comprising or expressing the alpha toxin epitope GNVTGDDTGKIGGLIG (SEQ ID NO: 17), can elicit an immune response to the alpha toxin epitope. This VLP can act together with the composition comprising an immunogen comprising / consisting essentially of the STAPD (SEQ ID NO: 1) epitope TNKDKNSTAPDDIGKNGK (SEQ ID NO:3), KDKNSTAPDDIGKNGKI (SEQ ID NO:4), STAPDDIGKNGKITKRT (SEQ ID NO:5), STAPD (SEQ ID NO: 1), STAPDD (SEQ ID NO:2). or an immunogenic equivalent thereof, wherein the immunogenic equivalent thereof is a peptide comprising / consisting essentially of an amino acid sequence at least 90% identical to TNKDKNSTAPDDIGKNGK (SEQ ID NO:3), KDKNSTAPDDIGKNGKI (SEQ ID NOT), STAPDDIGKNGKITKRT (SEQ ID NO: 5), STAPD (SEQ ID NO: 1), or STAPDD (SEQ ID NO:2), to treat a subject. In some aspects, this VLP can act together with the composition comprising an immunogen comprising / consisting essentially of the KEKR epitope KEKRNVTNKDKNSTAPD (SEQ ID NOTO), EHVDKSQQKEKRNVTNK (SEQ ID NO:8), KSQQKEKRNTNKDKNS (SEQ ID NOV), KEKR (SEQ ID NO:6), KDKR (SEQ ID NOT), orATTORNEY DOCKET NO. 37759.0626P1 an immunogenic equivalent thereof, wherein the immunogenic equivalent thereof is a peptide comprising / consisting essentially of an amino acid sequence at least 90% identical to KEKRNVTNKDKNSTAPD (SEQ ID NO: 10), EHVDKSQQKEKRNVTNK (SEQ ID NO:8), KSQQKEKRNTNKDKNS (SEQ ID NOV), KEKR (SEQ ID NO:6), or KDKR (SEQ ID NO:7) to treat a subject.4. Methods of Inducing an Immune Response

[0189] Disclosed are methods of inducing an immune response against Staphylococcus aureus in a subject, comprising administering to the subject one or more of the disclosed immunogenic compositions, peptides, nucleic acids, or VLPs, wherein the immune response targets Staphylococcus aureus leukotoxin LukH.

[0190] In some aspects, the immune response is LukH specific antibodies. In some aspects, the LukH specific antibodies are neutralizing antibodies.

[0191] Disclosed are methods of immunizing a subject against Staphylococcus aureus comprising administering to the subject one or more of the disclosed immunogenic compositions, peptides, nucleic acids, or VLPs, wherein the subject generates neutralizing antibodies to LukH.H. In some aspects, the neutralizing antibodies to LukH target the LukH of the LukGH complex. Thus, in some aspects, the neutralizing antibodies to LukH can be said to target LukGH. Administration

[0192] The immunogenic compositions, peptides, nucleic acids, or VLPs disclosed herein can be administered in a wide variety of therapeutic dosage forms in the conventional vehicles for topical, oral, systemic, local, and parenteral administration.

[0193] The route and regimen of administration will vary depending upon the population and the indication for vaccination and is to be determined by the skilled practitioner. For example, the immunogenic compositions, immunogens, or vaccines disclosed herein may be administered in such dosage forms for example as tablets, capsules (each including timed release and sustained release formulations), pills, powders, granules, elixirs, tinctures, solutions, suspensions, syrups, and emulsions. In some embodiments, the immunogenic compositions, immunogens, or vaccines disclosed herein may be administered by injection. Similarly, they may also be administered parentally, e.g., in intravenously (either by bolus or infusion methods), intraperitoneally, subcutaneously, topically with or without occlusion, or intramuscularly.

[0194] The administration may comprise an initial immunization or priming dose and at least one subsequent immunization or booster dose, following known standard immunizationATTORNEY DOCKET NO. 37759.0626P1 protocols. The boosting doses will be adequately spaced at such times where the levels of circulating antibody fell below a desired level. Boosting doses may consist of one or both of the peptides disclosed herein and may comprise alternative carriers and / or adjuvants. The booster dosage levels may be the same or different that those of the initial immunization dosage. Booster doses may be given at. for example. 1 week. 2 weeks. 3 weeks. 4 weeks, two months, three months, 6 months and / or a year later.

[0195] Alternatively, in some embodiments, the administration may comprise a single immunization (e.g., a single dose of the vaccine).

[0085] The specific dose levels may depend upon a variety of factors including the activity of the peptide, composition or vaccine, the age. body weight, general health, and diet of the subject, time of administration, and route of administration. For prophylaxis purposes, the amount of peptide in each dose is an amount which induces an immunoprotective response without significant adverse side effects. The dose range may be established empirically and may range from 10 micrograms to 500 micrograms for the single, priming and / or boosting doses.

[0196] The compositions, peptides, nucleic acids, or VLPs may be prepared, packaged, or sold in a form suitable for bolus administration or sold in unit dosage forms, such as in ampules or multi-dose containers containing a preservative.

[0197] A second therapy may be used in conjunction with the disclosed compositions, peptides, nucleic acids, or VLPs. The second therapy may be administration of an additional therapeutic agent or an additional vaccine. In some aspects, the second therapy is a known antibacterial agent, particularly a known S. aureus therapeutic. In some aspects, a second therapy can be formulated separately or with the disclosed compositions, peptides, nucleic acids, or VLPs.I. Kits

[0198] The compositions and materials described above as well as other materials can be packaged together in any suitable combination as a kit useful for performing, or aiding in the performance of, the disclosed method. It is useful if the kit components in a given kit are designed and adapted for use together in the disclosed method. For example disclosed are kits comprising one or more of the disclosed peptides, nucleic acids, vectors or VLPs and optionally, instructions for using the same.ExamplesATTORNEY DOCKET NO. 37759.0626P1A. Virus like particle vaccines targeting novel linear neutralizing epitopes in the LukH chain of the bicomponent leukotoxin LukGH from Staphylococcus aureus1. Materials and Methods i. Synthetic peptides

[0199] Multiple antigenic peptides (MAPs) displaying peptide sequences from LukGH, subunit G (GenBank # UVI99565.1) and subunit H (GenBank # NGG26803. 1) were synthesized commercially (Bio-Synthesis, Inc., Lewisville TX). For all studies employing MAPs, each construct was synthesized with the respective leukotoxin candidate B cell epitope sequence synthesized collinearly at the C-terminus of the T* helper T cell epitope, as well as separately with the P30 helper T cell epitope from tetanus toxin. Control MAPs used for this study consisted of the T* and P30 helper epitopes separately linked to irrelevant B cell sequences. ii. Animals and Vaccinations

[0200] For generation of rabbit antisera using MAPs, female New Zealand white (NZW) rabbits were immunized on day 0 with 300 pg of the respective MAP immunogens in an emulsion with CFA and were then boosted 3 times at two-week intervals with 150 pg of the respective MAP in an emulsion with IF A. Test bleeds were obtained two weeks after the fourth injection (8 weeks). For immunization of rabbits with VLPs, 300 pg of the respective VLP in an emulsion with IFA was administered s.c. on day 0, and each rabbit was then administered a second injection of 150 pg in IFA at week 5. Test bleeds were obtained 3 weeks after the second injection at week 8. iii. Construction and purification of Recombinant Chimeric WHc-Based VLPs

[0201] The recombinant WHc VLPs were moleculary constructed, validated, expressed and purified . Briefly, WHcAg VLPs were constructed by modifying pUC-FLw2 (Full-Length woodchuck) vector expressing the full-length WHcAg protein codon optimized for expression in E. coli essentially. The sequence for FLw2 matches the sequence translated from the woodchuck hepatitis virus core protein open reading frame (accession Ml 8752) and was cloned into a pUC19 vector in place of the multiple cloning site. For inserting heterologous B cell epitopes, EcoRI-XhoI restriction sites were engineered into the FLw2 open reading frame between amino acids 78 and 79 of the core protein gene. The engineered restnction sites add a Gly-Ile-Leu linker on the N-terminal side and a Leu linker on the C-terminal side of the inserted epitopes. Sequences from N-terminus of LukGH, subunit H (Genbank NGG26803.1) were cloned into the VLP gene using synthetic oligonucleotides comprising the desired epitope coding sequence and the appropriate engineered restriction sites. All WHcAg constructs were transformed into Alpha-Select competent E. coli (Bioline USA, Inc., Taunton, MA), andATTORNEY DOCKET NO. 37759.0626P1 selected clones were grown in Terrific Broth (Teknova, Hollster, CA). Clone-specific plasmid DNA wa purified by Zymo Zyppy™ Plasmid Miniprep Kit (Zymo Research, Irivine, CA) and correct sequences were confirmed by Sanger DNA sequencing (EuroFins MWG Operon USA, Louisville, KY). For purification of VLPs, cells were lysed using an EmulsiFlex-C3 (Avestin, Ottawa, ON. Canada) and lysate clarified by centrifugation. The Whc-Ag VLP particles were selectively precipitated with solid ammonium sulfate to approximately 45% saturation and the precipitates were collected by centrifugation at 20 x g. Precipitated VLPs were resuspended in minimum buffer (lOmM Tris, pH8, lOOmM NaCl). Purified VLPs were 0.2 pm sterile-filtered, characterized by custom ELISA for antigenicity, and by native agarose gel electrophoresis, SDS- PAGE and heat stability. iv. Preparation of Bacterial Supernatants

[0202] For preparation of bacterial supernatants for use in the HL60-based leukotoxin TNA, S. aureus strains were grown overnight (ON) in tryptic soy broth (TSB. Sigma Biochemicals, St. Louis, MO) at 37°C with shaking at 230 rpm in 200 ml Erlenmeyer flasks. ON cultures were diluted 1:100 and were expanded for 4 hours at 37°C and 230 rpm until mid-exponential phase (approximately 0.7 OD at 600 nm). Bacteria were pelleted by centrifugation at 14,000RPM for 4 minutes and supernatants were snap-frozen on dry ice and stored at -80C until use. v. Enzyme-linked immunosorbent assay

[0203] Antibody responses were assessed by ELISA. For analysis of antibodies specific for LukGH, wells of microtiter plates (Immulon 2, Thermo Labsystems, Franklin MA) were coated overnight at 4°C with 1 ug / well of recombinant LukGH (LukA-LukB, #0510-001, IBT Inc. Rockville, MD), in a 0.05 M carbonate buffer pH 9.5. Bound antibody was detected with secondary biotinylated antibody specific for rabbit IgG (Southern Biotechnology, Birmingham, AL) followed by streptavidin-alkaline phosphatase and 4-nitrophenylphosphate (Roche, Indianapolis, IN). Absorbance at 405 nm minus absorbance at 650 nm was determined using a microplate reader (Emax, Molecular Devices, Menlo Park, CA). Antibody titers were determined from serial two-fold dilutions of serum and represent the reciprocal dilution at the EC50 established using nonlinear regression to fit a variable slope sigmoidal equation to the serial dilution data using Prism vlO. The lower limit of quantitation for the ELISA was 16. vi. Toxin Neutralization Assay

[0204] The ability of antibody to block LukGH cytotoxicity in vitro was assessed using the HL60 cell line (CCL-240, ATCC. Manassas, VA). Briefly, HL60 cells were grown in culture in RPMI with 15% fetal bovine serum(FBS), penicillin-streptomycin and 50 pM 2ME (completeATTORNEY DOCKET NO. 37759.0626P1 medium) in a humidified 6.0 % CO2 incubator. For preparation of the cells for use in the leukotoxin TNA, HL60 cells were incubated for 2-3 days with complete medium containing DMSO at a final concentration of 1.5% volume: volume.

[0205] For each assay, rabbit sera in duplicate was serially diluted with complete medium containing 5% FBS in polypropylene round-bottom 96 well plates in a final volume of 50 microliters per well. Recombinant LukGH reagent (LukA-LukB, 0510-001, IBT. Rockville MD) was prepared at four times (4X) the final concentration, with the final concentration representing 3 to 4 multiples of the amount needed to kill 50% of the HL60 cells (Toxic dose 50% or TD50). Serially diluted rabbit antiserum was added to the LukGH toxin and the mixture was returned to the incubator for 30 minutes, after which time, 100,000 HL60 cells in 0. 1 ml of complete medium with 5% FBS were added to each well. Following a 2-hour incubation, 40 microliters of WST-8 reagent (Genscript USA Inc., Piscataway, NJ) containing a water-soluble tetrazolium salt and electron mediator was added, and the absorbance at 450 nm minus absorbance at 650 nm was determined for each plate approximately 18 hours later using a microplate reader.Neutralization ED50 (effective dilution at which 50% of cells are protected from cytotoxicity') titers were determined from serial two-fold dilutions of individual rabbit serum and represent the reciprocal dilution at the EC50 established using nonlinear regression to fit a variable slope sigmoidal equation to the serial dilution data using Prism 10. The standard TNA assay has a lower limit of quantification of 8. Samples with TNA titers below the lower limit of quantification were assigned a value of 4. Each TNA assay was validated by performance of a contemporaneous LukGH titration. For the analysis of peptide inhibition in the TNA, experimental serum samples were pre-incubated 1: 1 with 20 pM peptides for 30 minutes at room temperature prior to analysis in the TNA. vii. Statistical Analysis

[0206] For determination of ELISA and TNA ED50 titers, four-parameter nonlinear regression was used to fit variable slope sigmoidal equations to the serial dilution data. For all statistical analysis, ap value of < 0.05 was considered significant. All statistical analysis was performed using Prism 10.2. Results i. A neutralizing epitope in the N-terminal domain of LukH

[0207] Both AT and LukGH are PFTs and they share homolog)’ in the structure of their monomeric and assembled toxin forms (FIGS. 1 and 2). The monomeric polypeptides. LukGATTORNEY DOCKET NO. 37759.0626P1 and LukH, which together constitute the assembled heterodimer toxin, LukGH, could contain a neutralizing epitope in the N-terminal region akin to the NTD previously identified within the N-terminus of AT. Clustal analysis was performed which revealed alignment between two regions at the N-terminus of LukH and one region at the N-terminus of LukG, with the region in AT where the sequences comprising the NTD were delineated (FIG. 3). In an effort analogous to the one employed in identifying the NTD in AT, MAP constructs were synthesized spanning the sequences which could contain neutralizing epitopes in LukG and LukH (Table 1).

[0208] Table 1. Linear peptide sequences displayed in the respective MAP immunogens used to elicit humoral immunity specific for the LukGH monomers.

[0209] Each leukotoxin target sequence: LukH-Nterm, LukH*-Nterm and LukG-Nterm, was synthesized collinearly to the C-terminus of T*, a helper T cell epitope from Plasmodium falciparum as well as separately with the P30 helper T cell epitope from tetanus toxin, resulting in two separate MAPs for each B cell target sequence. Both of these helper T cell epitopes have been demonstrated, singularly and together, to be effective sources of linked T cell help in outbred rabbits for use in eliciting antibody responses towards putative B cell targets. Groups of rabbits (n=2) were immunized, using CFA for priming and IFA for boosting, with a mixture of two MAPs, each containing the same leukotoxin B cell target, but with the separate, heterologous helper T cell epitopes as described. After the fourth immunization, rabbit serum was analyzed by ELISA, and in the LukGH toxin neutralization assay (TNA) employed for these studies. As shown in FIG.4A, all rabbits developed Ab reactive with the immobilized LukGH heterodimer by ELISA, and one rabbit immunized with the LukH* MAP demonstrated neutralization in the TNA(FIG.4B). To confirm the specificity of the observed neutralization response in the sera of the rabbit immunized with the LukH* MAP, the TNA was repeated following pre-incubation of the sera w ith either the LukH-Nterm, LukH*-Nterm or the LukG- Nterm MAPs. As shown in FIG.5, both the LukH and LukH* MAPs completely inhibited the neutralization in the rabbit serum, while pre-incubation with the LukG-Nterm MAP had noATTORNEY DOCKET NO. 37759.0626P1 inhibitory effect. The results confirmed the specificity of the neutralizing antibody response and further, localized the likely neutralizing epitope to the sequence:TNKDKNSTAPDDIGKNGK (SEQ ID NO: 3), since this peptide sequence is shared by both LukH-Nterm and LukH*-Nterm MAPs (Table 1). ii. Mapping of the neutralizing epitope within the N-terminus of LukH.

[0210] To specifically delineate the neutralizing epitope identified through immunization of rabbits with the LukH*-Nterm MAP, a series of overlapping peptides were synthesized spanning the regions likely to contain the neutralizing epitope elicited by the LukH*-Nterm MAP sequence, while also extending to sequences N-terminal to this MAP sequence in the areas originally interrogated using the LukH-Nterm MAP (FIG.6). Like the original immunization strategy to elicit Ab against the N-terminal regions of LukH and LukG, each new B cell target sequence was synthesized collinearly with the T* helper T cell epitope in the MAP, and in a separate MAP, was synthesized collinear with the P30 helper T cell epitope. Groups of NZW rabbits (n=2) were then immunized 4 times at 2-week intervals with each MAP mixture, and sera was obtained from all rabbits at w eek 8, two-weeks after the fourth immunization, for analysis by ELISA and in the LukGH TNA. As shown in FIG.7A, all rabbits generated antibody which was immunoreactive with immobilized LukGH by ELISA, and immunization of rabbits with five of the MAP sequences elicited neutralizing antibody in the TNA (figure 7B). While the EH41 MAP elicited the highest levels of neutralization, it shares only 3 amino acids w ith the LukH*MAP, indicating there may be two separate neutralizing epitopes in the regions examined at the N-terminus of LukH. Indeed, using the wide-array of MAPs synthesized for this study, inhibition TNAs were performed to more precisely define the critical amino acids comprising the neutralizing epitope(s). The results of the inhibition TNAs revealed two distinct, nonoverlapping epitopes at the N-terminus of LukH: KEKR and STAPD (FIG.8). iii. A VLP displaying the STAPD epitope elicits neutralizing antibody

[0211] Though the use of MAPs as immunogens can be effective for eliciting antisera to identity' and delineate toxin neutralizing epitopes, they do not to-date have utility' for use in humans since MAPs typically require potent adjuvants like CFA for priming immunizations, and are not typically immunogenic when formulated with human-use adjuvants. Virus-like-particles (VLPs) were molecularly engineered, using the Woodchuck Hepatitis capsid (WHc) protein, displaying two iterations of the STAPD-containing peptide sequences. WHc VLPs display 240 copies of the target sequence at the surface of the VLP nanoparticle and have been show n to be a highly immunogenic platform for eliciting antibody responses against target sequences. Tw oATTORNEY DOCKET NO. 37759.0626P1 distinct VLPs, referred to as NP123 and NP125, each display distinct peptide sequences from LukH comprising the STAPD (SEQ ID NO: 1) neutralizing determinant, but the two VLPs vary in their amino acid margins outside the STAPD (SEQ ID NO: 1) motif (FIG.9). Each VLP was molecularly cloned and purified, and the results of the purifications are shown in FIG. 10. Following the purification of the two VLPs. groups of rabbits (n=4) were immunized 2 times at week 0 and week 5 with either the NP123 or the NP125 formulated in IFA for priming and boosting. Rabbit antisera was procured 3 weeks after the booster immunization (week 8) and was analyzed by ELISA for reactivity with immobilized LukGH, and for neutralization of LukGH in the TNA. As shown in FIG. 11 A, all rabbits immunized with either the NP123 or NP125 developed antibody reactive with immobilized LukGH, and all sera demonstrated the capacity to neutralize recombinant LukGH in the TNA(FIG. 1 IB). One rabbit immunized with the NP123, which had the highest antibody titer by ELISA, demonstrated a neutralization titer of over 40,000, which is ten times higher than the neutralization titer of a commercially-available, affinity-purified neutralizing pooled IgG positive control which was derived from immunizing multiple rabbits with full-length LukGH, thus highlighting the potential for an epitope-specific vaccine specific for LukH. Next it was evaluated whether the antisera raised to the NP123 could also neutralize native LukGH in the supernatant of S. aureus Newman, a historically important MSSA strain which produces large amounts of LukGH. and which was used in seminal studies to define the identity and role of this leukotoxin in S. aureus infections. As shown in FIG. 12, antisera from rabbits immunized w ith the NP123 were capable of neutralizing LukGH in the supernatant of S', aureus Newman. The results highlight the potential for targeting the STAPD neutralizing epitope displayed on the WHc VLP for the generation of high-titer neutralizing Ab specific for LukGH, a critical virulence factor for S. aureus infections in humans. iv. A VLP displaying the KEKR epitope elicits LukGH-specific antibody in rabbits

[0212] Since the results with MAPs enumerated two distinct neutralizing epitope motifs at the N-terminus of the LukH polypeptide: STAPD (SEQ ID NO: 1) and KEKR (SEQ ID NO:6), an initial VLP was molecularly constructed displaying the peptide sequence KEKR (SEQ ID NO:6) neutralizing motif, shown diagrammatically in FIG. 13. Following molecular cloning and purification of the KEKR-displaying VLP, a group of NZW rabbits (n=3) was immunized two times at weeks zero and five with the VLP 128 using IFA for priming and boosting. Week 8 antisera, obtained 3 weeks after the second immunization, was analyzed for reactivity with LukGH by ELISA. As shown in FIG.14A, all rabbits immunized with the VLP128 generated high-titer antibody reactive with immobilized LukGH.

[0213] As show n in FIG14B, when evaluated in the LukGH TNA, one rabbit demonstratedATTORNEY DOCKET NO. 37759.0626P1LukGH neutralization at levels equivalent to the commercially-available, affinity-purified neutralizing pooled IgG positive control which was derived from immunizing multiple rabbits with full-length LukGH. As with the results from the LukGH TNA with the STAPD VLPs NP123 and NP125, the results with the VLP128 highlight the potential for the KEKR-targeted epitope-specific VLPs for use in neutralizing LukGH in the treatment and prevention of S. aureus infections.3. Discussion

[0214] In the current study, whether a linear neutralizing epitope existed in the N-terminal regions of the LukG and LukH polypeptides which together comprise the leukotoxin heterodimer, LukGH, was evaluated. LukGH is a critical virulence factor for S. aureus infections in humans, that primarily acts through cytotoxicity of host innate and adaptive immune effector cells.

[0215] Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the method and compositions described herein. Such equivalents are intended to be encompassed by the following claims.

Claims

ATTORNEY DOCKET NO. 37759.0626P1CLAIMSWe claim:

1. A peptide, wherein the peptide comprises 5 to 30 amino acids, wherein the amino acid sequence comprises the amino acid sequence STAPD (SEQ ID NO: 1).

2. A peptide, wherein the peptide compnses 6 to 30 amino acids, wherein the amino acid sequence comprises the amino acid sequence STAPDD (SEQ ID NO:2).

3. The peptide of any one of claims 1-2, wherein the amino acid sequence is 90% identical to a portion of LukH.

4. The peptide of claim 3, wherein the amino acid sequence is 90% identical to amino acids 33-77 of LukH (SEQ ID NO. 11).

5. The peptide of any one of claims 1-4, wherein the amino acid sequence comprises an amino acid sequence at least 90% identical to the amino acid sequence TNKDKNSTAPDDIGKNGK (SEQ ID NO:3).

6. The peptide of any one of claims 1-4, wherein the amino acid sequence comprises an amino acid sequence at least 90% identical to the amino acid sequence KDKNSTAPDDIGKNGKI (SEQ ID NO:4).

7. The peptide of any one of claims 1-4, wherein the amino acid sequence comprises an amino acid sequence at least 90% identical to the amino acid sequence STAPDDIGKNGKITKRT (SEQ ID NO: 5).

8. The peptide of any one of claims 1-7, wherein the peptide further comprises the amino acid sequence KEKR (SEQ ID NO:6).

9. The peptide of claim 8. wherein the amino acid sequence comprises the amino acid sequence KEKRNVTNKDKNSTAPD (SEQ ID NO: 10).

10. A peptide, wherein the peptide comprises 4 to 30 amino acids, wherein the amino acid sequence comprises the amino acid sequence KEKR (SEQ ID NO: 6).

11. The peptide of claim 10, wherein the amino acid sequence is 90% identical to a portion of LukH (SEQ ID NO: 11).ATTORNEY DOCKET NO. 37759.0626P112. The peptide of claims 10-11, wherein the amino acid sequence is 90% identical to ammo acids 33-70 of LukH (SEQ ID NO: 11).

13. The peptide of any one of claims 10-12, wherein the amino acid sequence comprises an amino acid sequence at least 90% identical to the amino acid sequence EHVDKSQQKEKRNVTNK (SEQ ID NO: 8).

14. The peptide of any one of claims 10-13, wherein the amino acid sequence comprises an amino acid sequence at least 90% identical to the amino acid sequence KSQQKEKRNTNKDKNS (SEQ ID NO: 9).

15. The peptide of any one of claims 10-14, wherein the amino acid sequence further comprises the amino acid sequence STAPD (SEQ ID NO: 1) or STAPDD (SEQ ID NOT).

16. The peptide of claim 15, wherein the amino acid sequence comprises an amino acid sequence having the amino acid sequence KEKRNVTNKDKNSTAPD (SEQ ID NOTO).

17. A polypeptide comprising a first peptide and a second peptide, wherein the first peptide is any one of the peptides of claims 1-16, wherein the second peptide is helper T cell epitope.

18. A peptide consisting of the amino acid sequence STAPD (SEQ ID NOT), STAPDD (SEQ ID NOT), TNKDKNSTAPDDIGKNGK (SEQ ID NOT).KDKNSTAPDDIGKNGKI (SEQ ID NOT). STAPDDIGKNGKITKRT (SEQ ID NOT), KEKRNVTNKDKNSTAPD (SEQ ID NOTO), KEKR (SEQ ID NOT), EHVDKSQQKEKRNVTNK (SEQ ID NOT), or KSQQKEKRNTNKDKNS (SEQ ID NOT).

19. A peptide comprising or consisting essentially of an amino acid sequence at least 90% identical to the amino acid sequence STAPD (SEQ ID NOT), STAPDD (SEQ ID NOT), TNKDKNSTAPDDIGKNGK (SEQ ID NOT), KDKNSTAPDDIGKNGKI (SEQ ID NO:4), STAPDDIGKNGKITKRT (SEQ ID NOT), KEKRNVTNKDKNSTAPD (SEQ ID NOTO), KEKR (SEQ ID NO: 6).EHVDKSQQKEKRNVTNK (SEQ ID NOT), or KSQQKEKRNTNKDKNS (SEQ ID NOT).ATTORNEY DOCKET NO. 37759.0626P120. A modified viral capsid protein comprising a portion of LukH, wherein the portion of LukH comprises the amino acid sequence STAPD (SEQ ID NO: 1), KEKR (SEQ ID NO:6), or both.

21. The modified viral capsid protein of claim 20, wherein the modified viral capsid protein is a modified woodchuck heptatis virus capsid protein.

22. The modified viral capsid protein of any one of claims 20-21, wherein the portion of LukH is inserted within the modified viral capsid protein.

23. A virus-like particle (VLP) comprising 240 modified viral capsid proteins of claims 19-21.

24. An expression vector comprising one or more of the peptides of claims 1-19.

25. The expression vector of claim 24, wherein the one or more peptides of claims 1-19 are present in a viral capsid protein.

26. The expression vector of claim 25, wherein the modified viral capsid protein is one of claims 19-21.

27. A nucleic acid sequence encoding a LukH peptide.

28. The nucleic acid sequence of claim 27, wherein the LukH peptide comprises the amino acid sequence STAPD (SEQ ID NO:1) or STAPDD (SEQ ID NO:2).

29. The nucleic acid sequence of claim 27 or claim 28, wherein the LukH peptide is selected from the group consisting of the amino acid sequences of STAPD (SEQ ID NO: 1), STAPDD (SEQ ID NO:2), TNKDKNSTAPDDIGKNGK (SEQ ID NO:3), KDKNSTAPDD1GKNGKI (SEQ ID NO:4). STAPDD1GKNGKITKRT (SEQ ID NO:5), KEKRNVTNKDKNSTAPD (SEQ ID NOTO), KEKR (SEQ ID NO:6), EHVDKSQQKEKRNVTNK (SEQ ID NO: 8), or KSQQKEKRNTNKDKNS (SEQ ID NO:9).

30. The nucleic acid sequence of any of claims 27-29, wherein the nucleic acid sequence encodes the amino acid sequence of a polypeptide capable of forming a virus like particle.ATTORNEY DOCKET NO. 37759.0626P131. The nucleic acid sequence of claim 30. wherein the polypeptide capable of forming a virus like particle is a woodchuck hepatitis DNA virus core antigen.

32. The nucleic acid sequence of claim 31, wherein amino acid sequence of the woodchuck hepatitis DNA virus core antigen comprises an amino acid sequence having at least 70% identity to the amino acid sequence MDIDPYKEFGSSYQLLNFLPLDFFPDLNALVDTATALYEEELTGREHCSPHHT AIRQALVCWDELTKLIAWMSSNITSEQVRTIIVNHVNDTWGLKVRQSLWFHL SCLTFGQHTVQEFLVSFGVWIRTPAPYRPPNAPILSTLPEHTVIRRRGGARASR SPRRRTPSPRRRRSQSPRRRRSQSPSANC (SEQ ID NO: 12).

33. An expression vector comprising the nucleic acid of any of claims 27-32 in combination with a promoter.

34. A composition comprising any one of the peptides of claims 1-19, modified viral capsid proteins of claims 20-22, vims-like particle of claim 23, expression vectors of claims 24-26, or nucleic acid sequences of claims 27-32.

35. A composition comprising a peptide comprising or consisting essentially of the amino acid sequence TNKDKNSTAPDDIGKNGK (SEQ ID NO: 3), KDKNSTAPDDIGKNGKI (SEQ ID NO: 4), STAPDDIGKNGKITKRT (SEQ ID NO:5) or a peptide equivalent thereof, wherein the peptide equivalent thereof is a peptide comprising or consisting essentially of an amino acid sequence at least 90% identical to the amino acid sequence TNKDKNSTAPDDIGKNGK (SEQ ID NO:3), DKNSTAPDDIGKNGKI (SEQ ID NO: 4), STAPDDIGKNGKITKRT (SEQ ID NO:5).

36. The composition of claim 35, comprising a peptide consisting essentially of the amino acid sequence STAPD (SEQ ID NO:1), TNKDKNSTAPDDIGKNGK (SEQ ID NO:3), KDKNSTAPDDIGKNGKI (SEQ ID NO:4), STAPDDIGKNGKITKRT (SEQ ID NO:5)or an immunogenic equivalent thereof, wherein the immunogenic equivalent thereof is a peptide comprising / consi sting essentially of an amino acid sequence at least 90% identical to the amino acid sequence TNKDKNSTAPDDIGKNGK (SEQ ID NO: 3), KDKNSTAPDDIGKNGKI (SEQ ID NO: 4), STAPDDIGKNGKITKRT (SEQ ID NO:5).ATTORNEY DOCKET NO. 37759.0626P137. The composition of any of claims 35-36, further comprising an adjuvant.

38. The composition of any of claims 35-37, further comprising one or more additional STAPD (SEQ ID NO: 1) peptides.

39. The composition of any of claims 35-38, further comprising one or more peptides comprising the amino acid sequence KEKR (SEQ ID NO:6).

40. The composition of claim 39. wherein the one or more of the peptides comprising the ammo acid sequence KEKR (SEQ ID NO: 6) is EHVDKSQQKEKRNVTNK (SEQ ID NO:8), KSQQKEKRNTNKDKNS (SEQ ID NO:9) or an immunogenic equivalent thereof, wherein the immunogenic equivalent thereof is a peptide comprising / consisting essentially of an amino acid sequence at least 90% identical to the ammo acid sequence EHVDKSQQKEKRNVTNK (SEQ ID NO: 8) or KSQQKEKRNTNKDKNS (SEQ ID NO: 9).

41. A composition comprising a peptide comprising or consisting essentially of a peptide comprising the amino acid sequence EHVDKSQQKEKRNVTNK (SEQ ID NO: 8). KSQQKEKRNTNKDKNS (SEQ ID NO:9) or a peptide equivalent thereof, wherein the peptide equivalent thereof is a peptide comprising or consisting essentially of an amino acid sequence at least 90% identical to the amino acid sequence EHVDKSQQKEKRNVTNK (SEQ ID NO: 8) or KSQQKEKRNTNKDKNS (SEQ ID NO:9).

42. The composition of claim 41. comprising a peptide consisting essentially of a peptide comprising the amino acid sequence EHVDKSQQKEKRNVTNK (SEQ ID NO: 8), KSQQKEKRNTNKDKNS (SEQ ID NO:9) or a peptide equivalent thereof, wherein the peptide equivalent thereof is a peptide comprising / consisting essentially of an amino acid sequence at least 90% identical to the amino acid sequence EHVDKSQQKEKRNVTNK (SEQ ID NO: 8) or KSQQKEKRNTNKDKNS (SEQ ID NO:9).

43. The composition of any one of claims 41-42, further comprising an adjuvant.

44. The composition of any one of claims 41-43. further comprising one or more additional peptides comprising the amino acid sequence KEKR (SEQ ID NO:6).ATTORNEY DOCKET NO. 37759.0626P145. The composition of any one of claims 41-44. further comprising one or more peptides comprising the amino acid sequence STAPD (SEQ ID NO:1).

46. A method of neutralizing the cytotoxicity of leukotoxin LukH in a subject infected with Staphylococcus aureus comprising administering to the subj ect any one of the peptides of claims 1-19, virus-like particle of claim 23, expression vectors of claims 24-26, nucleic acid sequences of claims 27-32, or compositions of claims 33-45, wherein the subject generates neutralizing antibodies (nAbs) against the LukH.

47. A method of treating a subject infected with Staphylococcus aureus comprising administering to the subject any one of the peptides of claims 1-19, virus-like particle of claim 23, expression vectors of claims 24-26, nucleic acid sequences of claims 27- 32, or compositions of claims 33-45.

48. The method of claim 47, wherein the subject generates neutralizing antibodies (nAbs) against Staphylococcus aureus leukotoxin LukH.

49. The method of claim 48, wherein the nAbs neutralize the cytotoxicity of the LukH.

50. The method of any one of claims 47-49, further comprising administering one or more antibiotics.

51. The method of claim 50, wherein the antibiotic is tetracycline, doxicycline, minocycline, trimethoprim-sulfamethoxazole, rifampin, clindamycin, vancomycin, linezolid, daptomycin, tigecycline, telavancin, dalbavancin, oritavancin. ceftobiprole, mupirocin or iclaprim.

52. The method of any one of claims 47-51, further comprising administering one or more antibodies that are specific for any Staphylococcus aureus epitope or VLP expressing any Staphylococcus aureus epitope.

53. The method of claim 52, wherein the Staphylococcus aureus epitope is an alpha toxin epitope.

54. The method of claim 53, wherein the alpha toxin epitope comprises the amino acid sequence GNVTGDDTGKIGGLIG (SEQ ID NO: 17).ATTORNEY DOCKET NO. 37759.0626P155. A method of inducing an immune response against Staphylococcus aureus in a subject, comprising administering to the subject any one of peptides of claims 1-19, virus-like particle of claim 23, expression vectors of claims 24-26, nucleic acid sequences of claims 27-32, or compositions of claims 33-45, wherein the immune response targets Staphylococcus aureus leukotoxin LukH.

56. The method of claim 55, wherein the immune response is LukH specific antibodies.

57. The method of claim 56, wherein the LukH specific antibodies are neutralizing antibodies.

58. A method of immunizing a subject against Staphylococcus aureus comprising administering to the subject any one of the peptides of claims 1-19, virus-like particle of claim 23. expression vectors of claims 24-26, nucleic acid sequences of claims 27- 32, or compositions of claims 33-45, wherein the subject generates neutralizing antibodies to LukH.

59. The method of any one of claims 55-58, further comprising administering one or more antibodies that are specific for any Staphylococcus aureus epitope or VLP expressing any Staphylococcus aureus epitope.

60. The method of claim 59, wherein the Staphylococcus aureus epitope is an alpha toxin epitope.

61. The method of claim 60, wherein the alpha toxin epitope comprises the amino acid sequence GNVTGDDTGKIGGLIG (SEQ ID NO: 17).

62. A method of preventing a Staphylococcus aureus infection in a subject comprising administering to the subject any one of the peptides of claims 1-19, virus-like particle of claim 23. expression vectors of claims 24-26, nucleic acid sequences of claims 27- 32, or compositions of claims 33-45, wherein the subject generates an immune response to LukH63. The method of claim 62, wherein the immune response comprises neutralizing antibodies to LukH.ATTORNEY DOCKET NO. 37759.0626P164. The method of any one of claims 62-63, further comprising administering one or more antibodies that are specific for any Staphylococcus aureus epitope or VLP expressing any Staphylococcus aureus epitope.

65. The method of claim 64, wherein the Staphylococcus aureus epitope is an alpha toxin epitope.

66. The method of claim 65, wherein the alpha toxin epitope comprises the amino acid sequence GNVTGDDTGKIGGLIG (SEQ ID NO: 17).

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