Allergy vaccine platform based on supramolecular materials

Abstract

Embodiments are directed to a conjugate peptide including a self-assembling peptide and at least one allergen epitope. Allergen epitopes may include peanut allergens. The conjugate peptide may self-assemble into a nanofiber or fibril. Compositions including the conjugate peptide may be used to treat allergies.

Description

Docket No.028193-0048-WO01 / 8355 ALLERGY VACCINE PLATFORM BASED ON SUPRAMOLECULAR MATERIALS CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisional Patent Application No. 63 / 612,664, filed December 20, 2023, and U.S. Provisional Patent Application No. 63 / 655,650, filed June 4, 2024, each of which is incorporated herein by reference in its entirety. STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH

[0002] This invention was made with government support under grant 5R21AI164740 awarded by the National Institutes of Health (NIH) National Institute of Allergy and Infectious Diseases (NIAID). The government has certain rights in the invention. FIELD

[0003] Embodiments of this invention are directed generally to biology, medicine, and immunology. Certain aspects are directed to peptide conjugates and nanofibers and their use in inducing an allergen-specific IgG response with a reduced systemic allergic response. INTRODUCTION

[0004] Allergic disease incidence is steadily rising, yet allergen-desensitizing treatment options remain limited. Allergen immunotherapies (AIT) are the most clinically advanced treatment modalities but can require life-long dosing and suffer from frequent adverse events including instances of systemic anaphylaxis, leading to poor patient compliance and high cost. Although the therapeutic mechanism(s) of AIT remain incompletely understood, it is generally appreciated that an increase in allergen specific IgG (sIgG) antibody is a major mechanistic component. Indeed, recent development of therapeutic sIgG monoclonal antibody therapies suggest that sIgG can achieve significant allergen desensitization.

[0005] Type I hypersensitivity reactions are primarily driven by mast cells and basophils,which capture allergen-specific IgE antibody (sIgE) via the high affinity IgE receptor (Fc RI).Allergen exposure triggers Fc RI crosslinking and rapid cellular activation, culminating in therelease of effector molecules that drive allergic symptoms. sIgG facilitates desensitization through two pathways, both of which act to limit activation of allergic effector cells: (1) sIgG directly neutralizes allergen by competing with sIgE for access to allergen epitopes and (2) once incorporated into allergen immune complexes, sIgG engages the inhibitory Fc receptor(Fc RIIb) on effector cells thereby inhibiting cellular activation through Fc RI.Docket No.028193-0048-WO01 / 8355

[0006] Given the desensitizing nature of sIgG and the efficacy of sIgG monoclonal antibody therapeutics, allergen B cell vaccines are being studied. These approaches aim to exploit vaccine technologies to generate robust and long-lived sIgG responses with only a few doses, thus addressing major disadvantages of AIT. Both AIT and allergen vaccines work, in part, by activating allergen-specific B cells to produce immunoglobulin G (IgG) antibodies that can neutralize allergen and prevent IgE-mediated recognition of allergen by mast cells. However, like AIT, allergen vaccines are often highly reactogenic in allergic patients, hampering their use in therapeutic settings. Hypoallergenic allergen vaccine immunogens that do not activate mast cells when used to immunize allergic patients have been considered. Typically, these strategies involve the removal, disruption, or masking of IgE-reactive B cell epitopes from the allergen immunogen to avoid Type I hypersensitivity reactions. However, these epitopes are targets for vaccine-elicited IgG, and immunogenicity and desensitization can be negatively impacted by their omission. Overcoming these barriers for the clinical translation of allergen vaccines is a major challenge. SUMMARY

[0007] In an aspect, provided herein is an allergen conjugate peptide. The allergen conjugate peptide may include (i) a self-assembling peptide comprising a polypeptide having the amino acid sequence of bXXXb (SEQ ID NO: 1, wherein X is independently any amino acid and b is independently any positively charged amino acid), QQKFQFQFEQQ (SEQ ID NO: 12), FKFEFKFE (SEQ ID NO: 14), KFQFQFE (SEQ ID NO: 15), QQRFQFQFEQQ (SEQ ID NO: 16), QQRFQWQFEQQ (SEQ ID NO: 17), FEFEFKFKFEFEFKFK (SEQ ID NO: 18), QQRFEWEFEQQ (SEQ ID NO: 19), QQXFXWXFQQQ (SEQ ID NO: 20, where X is ornithine), FKFEFKFEFKFE (SEQ ID NO: 21), FKFQFKFQFKFQ (SEQ ID NO: 22), AEAKAEAKAEAKAEAK (SEQ ID NO: 23), AEAEAKAKAEAEAKAK (SEQ ID NO: 24), AEAEAEAEAKAKAKAK (SEQ ID NO: 25), RADARADARADARADA (SEQ ID NO: 26), RARADADARARADADA (SEQ ID NO: 27), SGRGYBLGGQGAGAAAAAGGAGQGGYGGLGSQG (SEQ ID NO: 28), EWEXEXEXEX (SEQ ID NO: 29, where X is Val, Ala, Ser, or Pro), WKXKXKXKXK (SEQ ID NO: 30, where X is Val, Ala, Ser, or Pro), KWKVKVKVKVKVKVK (SEQ ID NO: 31, where X is Val, A, Ser, or Pro), LLLLKKKKKKKKLLLL (SEQ ID NO: 32), VKVKVKVKVDPPTKVKVKVKV (SEQ ID NO: 33), VKVKVKVKVDPPTKVKTKVKV (SEQ ID NO: 34), KVKVKVKVKDPPSVKVKVKVK (SEQ ID NO: 35), VKVKVKVKVDPPSKVKVKVKV (SEQ ID NO: 36), VKVKVKTKVDPPTKVKTKVKV (SEQ ID NO: 37), QQKFxFQFEQQ (SEQ ID NO: 38, wherein x is Glu, Asp, or Asn), QQKFQxQFEQQ (SEQ ID NO: 39, wherein x is Trp or Tyr), or QQKFQFxFEQQ (SEQ ID NO: 40, wherein x is Glu, Asp, or Asn); and (ii) at least oneDocket No.028193-0048-WO01 / 8355 allergen epitope conjugated to a terminus of the self-assembling peptide. In some embodiments, the allergen epitope is a B cell allergen epitope. In some embodiments, the allergen epitope is IgE-reactive. In some embodiments, the allergen epitope comprises a peanut allergen epitope. In some embodiments, the peanut allergen epitope is from peanut allergen protein Ara h 1, Ara h 3, or Ara h 7, or Ara h 2. In some embodiments, the peanut allergen epitope comprises AH1b, AH3a, AH7a, AH1a, AH2a, or AH2b, or a combination thereof. In some embodiments, the peanut allergen epitope comprises AH1b, AH3a, or AH7a, or a combination thereof. In some embodiments, each self-assembling peptide forms a beta sheet and comprises a polypeptide having an amino acid sequence selected from QQKFQFQFEQQ (SEQ ID NO: 12), FKFEFKFE (SEQ ID NO: 14), KFQFQFE (SEQ ID NO: 15), QQRFQFQFEQQ (SEQ ID NO: 16), QQRFQWQFEQQ (SEQ ID NO: 17), FEFEFKFKFEFEFKFK (SEQ ID NO: 18), QQRFEWEFEQQ (SEQ ID NO: 19), QQXFXWXFQQQ (SEQ ID NO: 20, where X is ornithine), FKFEFKFEFKFE (SEQ ID NO: 21), FKFQFKFQFKFQ (SEQ ID NO: 22), AEAKAEAKAEAKAEAK (SEQ ID NO: 23), AEAEAKAKAEAEAKAK (SEQ ID NO: 24), AEAEAEAEAKAKAKAK (SEQ ID NO: 25), RADARADARADARADA (SEQ ID NO: 26), RARADADARARADADA (SEQ ID NO: 27), SGRGYBLGGQGAGAAAAAGGAGQGGYGGLGSQG (SEQ ID NO: 28), EWEXEXEXEX (SEQ ID NO: 29, where X is Val, Ala, Ser, or Pro), WKXKXKXKXK (SEQ ID NO: 30, where X is Val, Ala, Ser, or Pro), KWKVKVKVKVKVKVK (SEQ ID NO: 31, where X is Val, A, Ser, or Pro), LLLLKKKKKKKKLLLL (SEQ ID NO: 32), VKVKVKVKVDPPTKVKVKVKV (SEQ ID NO: 33), VKVKVKVKVDPPTKVKTKVKV (SEQ ID NO: 34), KVKVKVKVKDPPSVKVKVKVK (SEQ ID NO: 35), VKVKVKVKVDPPSKVKVKVKV (SEQ ID NO: 36), VKVKVKTKVDPPTKVKTKVKV (SEQ ID NO: 37), QQKFxFQFEQQ (SEQ ID NO: 38, wherein x is Glu, Asp, or Asn), QQKFQxQFEQQ (SEQ ID NO: 39, wherein x is Trp or Tyr), and QQKFQFxFEQQ (SEQ ID NO: 40, wherein x is Glu, Asp, or Asn). In some embodiments, each self-assembling peptide forms an alpha-helix and comprises a polypeptide having an amino acid sequence of bXXXb (SEQ ID NO: 1), wherein X is independently any amino acid and b is independently any positively charged amino acid. In some embodiments, the self-assembling peptide comprises the sequence QQKFQFQFEQQ(SEQ ID NO: 12) or Ac-QQKFQFQFEQQ-NH2(SEQ ID NO: 13) or bXXXb (SEQ ID NO: 1,wherein X is independently any amino acid and b is independently any positively charged amino acid). In some embodiments, b is independently selected from Arg and Lys. In some embodiments, bXXXb (SEQ ID NO: 1) is RAYAR (SEQ ID NO: 2) or KAYAK (SEQ ID NO: 3). In some embodiments, the self-assembling peptide comprises an amino acid sequenceof ZnbXXXbZm(SEQ ID NO: 5), wherein b is independently any positively charged aminoacid, Z is independently any amino acid, X is independently any amino acid, n is an integerDocket No.028193-0048-WO01 / 8355 from 0 to 20, and m is an integer from 0 to 20. In some embodiments, the self-assembling peptide comprises an amino acid sequence selected from QARILEADAEILRAYARILEAHAEILRAQ (Coil29, SEQ ID NO: 6), or QAKILEADAEILKAYAKILEAHAEILKAQ (SEQ ID NO: 7), or ADAEILRAYARILEAHAEILRAQ(SEQ ID NO: 8), or Ac-QARILEADAEILRAYARILEAHAEILRAQ-NH2(SEQ ID NO: 9), or Ac-QAKILEADAEILKAYAKILEAHAEILKAQ-NH2(SEQ ID NO: 10), or Ac-ADAEILRAYARILEAHAEILRAQ-NH2(SEQ ID NO: 11), or QAEILRAYARILEAQ (SEQ IDNO: 101), or QAEILRAYARILEAHAEILKAQ (SEQ ID NO: 102), or QAEILRAYARILEADAKILEAHAEILKAQ (SEQ ID NO: 103), or QAEILRAYARILEADAEILKAQAKILEAHAEILKAQ (SEQ ID NO: 104). In some embodiments, the self-assembling peptide forms a beta sheet and the at least one allergen epitope is attached to the C-terminus or the N-terminus of the self-assembling peptide or a combination thereof, or the self-assembling peptide forms an alpha-helix and the at least one allergen epitope is attached to the N-terminus of the self-assembling peptide. In some embodiments, 1 to 10 allergen epitopes are attached to the C-terminus or the N-terminus of the self-assembling peptide. In some embodiments, the self-assembling peptide forms a beta sheet and the allergen conjugate peptide further includes (iii) a PEG molecule or a PAS peptide conjugated to the self-assembling peptide. In some embodiments, the PAS peptide comprises a sequence of Pro-Ala-Ser or comprises the sequence of ASPAAPAPASPAAPAPSAPA (SEQ ID NO: 97), or a peptide having at least 80%, 85%, 90%, or 95% identity thereto. In some embodiments, the PEG molecule comprises PEG- 2000. In some embodiments, the PEG molecule or the PAS peptide is conjugated to the self-assembling peptide at the same or the opposite terminus from where the allergen epitope is attached. In some embodiments, the allergen conjugate peptide further includes (iv) at least one linker. In some embodiments, the at least one linker is between the at least one allergen epitope and the self-assembling peptide. In some embodiments, the at least one linker comprises a first linker between the at least one allergen epitope and the self- assembling peptide, and a second linker between the PEG molecule or the PAS peptide and the self-assembling peptide. In some embodiments, the at least one linker comprises SEQID NO: 83 (SGSG), SEQ ID NO: 84 ((Ser-Gly)2), SEQ ID NO: 85 (CCCCSGSG), SEQ IDNO: 86 (Gnwherein n is an integer from 1 to 10), SEQ ID NO: 87 (GSGS), SEQ ID NO: 88 (SSSS), SEQ ID NO: 89 (GGGS), SEQ ID NO: 90 (GGC), SEQ ID NO: 91 ((GGC)8), SEQID NO: 92 ((G4S)3), SEQ ID NO: 93 (KSGSG), SEQ ID NO: 94 (KKSGSG), SEQ ID NO: 95(EAAAK)2, or SEQ ID NO: 96 (GGAAY).Docket No.028193-0048-WO01 / 8355

[0008] In another aspect, provided herein is a nanofiber comprising a plurality of the allergen conjugate peptide as detailed herein, wherein the plurality of allergen conjugate peptides self-assemble into the nanofiber. In another aspect, provided herein is a nanofiber comprising: (i) at least one allergen conjugate peptide as detailed herein; and at least one of a peptide selected from a (ii) T-cell epitope-conjugate peptide, a (iii-a) PEG conjugate peptide, a (iii-b) PAS conjugate peptide, and a (iv) plain self-assembling peptide, or a combination thereof, (ii) wherein the T-cell epitope-conjugate peptide comprises: a self- assembling peptide comprising a polypeptide having the amino acid sequence of QQKFQFQFEQQ (SEQ ID NO: 12), bXXXb (SEQ ID NO: 1, wherein X is independently any amino acid and b is independently any positively charged amino acid), FKFEFKFE (SEQ ID NO: 14), KFQFQFE (SEQ ID NO: 15), QQRFQFQFEQQ (SEQ ID NO: 16), QQRFQWQFEQQ (SEQ ID NO: 17), FEFEFKFKFEFEFKFK (SEQ ID NO: 18), QQRFEWEFEQQ (SEQ ID NO: 19), QQXFXWXFQQQ (SEQ ID NO: 20, where X is ornithine), FKFEFKFEFKFE (SEQ ID NO: 21), FKFQFKFQFKFQ (SEQ ID NO: 22), AEAKAEAKAEAKAEAK (SEQ ID NO: 23), AEAEAKAKAEAEAKAK (SEQ ID NO: 24), AEAEAEAEAKAKAKAK (SEQ ID NO: 25), RADARADARADARADA (SEQ ID NO: 26), RARADADARARADADA (SEQ ID NO: 27), SGRGYBLGGQGAGAAAAAGGAGQGGYGGLGSQG (SEQ ID NO: 28), EWEXEXEXEX (SEQ ID NO: 29, where X is Val, Ala, Ser, or Pro), WKXKXKXKXK (SEQ ID NO: 30, where X is Val, Ala, Ser, or Pro), KWKVKVKVKVKVKVK (SEQ ID NO: 31, where X is Val, A, Ser, or Pro), LLLLKKKKKKKKLLLL (SEQ ID NO: 32), VKVKVKVKVDPPTKVKVKVKV (SEQ ID NO: 33), VKVKVKVKVDPPTKVKTKVKV (SEQ ID NO: 34), KVKVKVKVKDPPSVKVKVKVK (SEQ ID NO: 35), VKVKVKVKVDPPSKVKVKVKV (SEQ ID NO: 36), VKVKVKTKVDPPTKVKTKVKV (SEQ ID NO: 37), QQKFxFQFEQQ (SEQ ID NO: 38, wherein x is Glu, Asp, or Asn), QQKFQxQFEQQ (SEQ ID NO: 39, wherein x is Trp or Tyr), and QQKFQFxFEQQ (SEQ ID NO: 40, wherein x is Glu, Asp, or Asn); and at least one T- cell epitope conjugated to a terminus of the self-assembling peptide, wherein the at least one T-cell epitope is selected from PADRE and VAC, and wherein PADRE comprises a polypeptide having the amino acid sequence of aKXVAAWTLKAa (SEQ ID NO: 99, wherein “X” comprises cyclohexylalanine and “a” comprises D-alanine), and wherein VAC comprises a polypeptide having the amino acid sequence of QLVFNSISARALKAY (SEQ ID NO: 100), (iii-a) wherein the PEG conjugate peptide comprises: a self-assembling peptide comprising a polypeptide having the amino acid sequence of QQKFQFQFEQQ (SEQ ID NO: 12), bXXXb (SEQ ID NO: 1, wherein X is independently any amino acid and b is independently any positively charged amino acid), FKFEFKFE (SEQ ID NO: 14), KFQFQFE (SEQ ID NO: 15), QQRFQFQFEQQ (SEQ ID NO: 16), QQRFQWQFEQQ (SEQ ID NO: 17), FEFEFKFKFEFEFKFK (SEQ ID NO: 18), QQRFEWEFEQQ (SEQ ID NO: 19),Docket No.028193-0048-WO01 / 8355 QQXFXWXFQQQ (SEQ ID NO: 20, where X is ornithine), FKFEFKFEFKFE (SEQ ID NO: 21), FKFQFKFQFKFQ (SEQ ID NO: 22), AEAKAEAKAEAKAEAK (SEQ ID NO: 23), AEAEAKAKAEAEAKAK (SEQ ID NO: 24), AEAEAEAEAKAKAKAK (SEQ ID NO: 25), RADARADARADARADA (SEQ ID NO: 26), RARADADARARADADA (SEQ ID NO: 27), SGRGYBLGGQGAGAAAAAGGAGQGGYGGLGSQG (SEQ ID NO: 28), EWEXEXEXEX (SEQ ID NO: 29, where X is Val, Ala, Ser, or Pro), WKXKXKXKXK (SEQ ID NO: 30, where X is Val, Ala, Ser, or Pro), KWKVKVKVKVKVKVK (SEQ ID NO: 31, where X is Val, A, Ser, or Pro), LLLLKKKKKKKKLLLL (SEQ ID NO: 32), VKVKVKVKVDPPTKVKVKVKV (SEQ ID NO: 33), VKVKVKVKVDPPTKVKTKVKV (SEQ ID NO: 34), KVKVKVKVKDPPSVKVKVKVK (SEQ ID NO: 35), VKVKVKVKVDPPSKVKVKVKV (SEQ ID NO: 36), VKVKVKTKVDPPTKVKTKVKV (SEQ ID NO: 37), QQKFxFQFEQQ (SEQ ID NO: 38, wherein x is Glu, Asp, or Asn), QQKFQxQFEQQ (SEQ ID NO: 39, wherein x is Trp or Tyr), and QQKFQFxFEQQ (SEQ ID NO: 40, wherein x is Glu, Asp, or Asn), and at least one PEG molecule conjugated to a terminus of the self-assembling peptide, (iii-b) wherein the PAS conjugate peptide comprises: a self-assembling peptide comprising a polypeptide having the amino acid sequence of QQKFQFQFEQQ (SEQ ID NO: 12), bXXXb (SEQ ID NO: 1, wherein X is independently any amino acid and b is independently any positively charged amino acid), FKFEFKFE (SEQ ID NO: 14), KFQFQFE (SEQ ID NO: 15), QQRFQFQFEQQ (SEQ ID NO: 16), QQRFQWQFEQQ (SEQ ID NO: 17), FEFEFKFKFEFEFKFK (SEQ ID NO: 18), QQRFEWEFEQQ (SEQ ID NO: 19), QQXFXWXFQQQ (SEQ ID NO: 20, where X is ornithine), FKFEFKFEFKFE (SEQ ID NO: 21), FKFQFKFQFKFQ (SEQ ID NO: 22), AEAKAEAKAEAKAEAK (SEQ ID NO: 23), AEAEAKAKAEAEAKAK (SEQ ID NO: 24), AEAEAEAEAKAKAKAK (SEQ ID NO: 25), RADARADARADARADA (SEQ ID NO: 26), RARADADARARADADA (SEQ ID NO: 27), SGRGYBLGGQGAGAAAAAGGAGQGGYGGLGSQG (SEQ ID NO: 28), EWEXEXEXEX (SEQ ID NO: 29, where X is Val, Ala, Ser, or Pro), WKXKXKXKXK (SEQ ID NO: 30, where X is Val, Ala, Ser, or Pro), KWKVKVKVKVKVKVK (SEQ ID NO: 31, where X is Val, A, Ser, or Pro), LLLLKKKKKKKKLLLL (SEQ ID NO: 32), VKVKVKVKVDPPTKVKVKVKV (SEQ ID NO: 33), VKVKVKVKVDPPTKVKTKVKV (SEQ ID NO: 34), KVKVKVKVKDPPSVKVKVKVK (SEQ ID NO: 35), VKVKVKVKVDPPSKVKVKVKV (SEQ ID NO: 36), VKVKVKTKVDPPTKVKTKVKV (SEQ ID NO: 37), QQKFxFQFEQQ (SEQ ID NO: 38, wherein x is Glu, Asp, or Asn), QKFQxQFEQQ (SEQ ID NO: 39, wherein x is Trp or Tyr), and QQKFQFxFEQQ (SEQ ID NO: 40, wherein x is Glu, Asp, or Asn); and at least one PAS peptide conjugated to a terminus of the self-assembling peptide, wherein the PAS peptide comprises a sequence of Pro-Ala-Ser or comprises the sequence of ASPAAPAPASPAAPAPSAPA (SEQ ID NO: 97), or a peptide having at least 80%, 85%, 90%, or 95% identity thereto, and (iv) wherein the plain self-assembling peptide comprises aDocket No.028193-0048-WO01 / 8355 polypeptide having the amino acid sequence of bXXXb (SEQ ID NO: 1, wherein X is independently any amino acid and b is independently any positively charged amino acid), QQKFQFQFEQQ (SEQ ID NO: 12), FKFEFKFE (SEQ ID NO: 14), KFQFQFE (SEQ ID NO: 15), QQRFQFQFEQQ (SEQ ID NO: 16), QQRFQWQFEQQ (SEQ ID NO: 17), FEFEFKFKFEFEFKFK (SEQ ID NO: 18), QQRFEWEFEQQ (SEQ ID NO: 19), QQXFXWXFQQQ (SEQ ID NO: 20, where X is ornithine), FKFEFKFEFKFE (SEQ ID NO: 21), FKFQFKFQFKFQ (SEQ ID NO: 22), AEAKAEAKAEAKAEAK (SEQ ID NO: 23), AEAEAKAKAEAEAKAK (SEQ ID NO: 24), AEAEAEAEAKAKAKAK (SEQ ID NO: 25), RADARADARADARADA (SEQ ID NO: 26), RARADADARARADADA (SEQ ID NO: 27), SGRGYBLGGQGAGAAAAAGGAGQGGYGGLGSQG (SEQ ID NO: 28), EWEXEXEXEX (SEQ ID NO: 29, where X is Val, Ala, Ser, or Pro), WKXKXKXKXK (SEQ ID NO: 30, where X is Val, Ala, Ser, or Pro), KWKVKVKVKVKVKVK (SEQ ID NO: 31, where X is Val, A, Ser, or Pro), LLLLKKKKKKKKLLLL (SEQ ID NO: 32), VKVKVKVKVDPPTKVKVKVKV (SEQ ID NO: 33), VKVKVKVKVDPPTKVKTKVKV (SEQ ID NO: 34), KVKVKVKVKDPPSVKVKVKVK (SEQ ID NO: 35), VKVKVKVKVDPPSKVKVKVKV (SEQ ID NO: 36), VKVKVKTKVDPPTKVKTKVKV (SEQ ID NO: 37), QQKFxFQFEQQ (SEQ ID NO: 38, wherein x is Glu, Asp, or Asn), QQKFQxQFEQQ (SEQ ID NO: 39, wherein x is Trp or Tyr), or QQKFQFxFEQQ (SEQ ID NO: 40, wherein x is Glu, Asp, or Asn). In some embodiments, the cyclohexylalanine comprises D-alanine. In some embodiments, the T-cell epitope- conjugate peptide further comprises a linker between the T-cell epitope and the self- assembling peptide. In some embodiments, the linker comprises an amino acid sequenceselected from SEQ ID NO: 83 (SGSG), SEQ ID NO: 84 ((Ser-Gly)2), SEQ ID NO: 85(CCCCSGSG), SEQ ID NO: 86 (Gnwherein n is an integer from 1 to 10), SEQ ID NO: 87(GSGS), SEQ ID NO: 88 (SSSS), SEQ ID NO: 89 (GGGS), SEQ ID NO: 90 (GGC), SEQ IDNO: 91 ((GGC)8), SEQ ID NO: 92 ((G4S)3), SEQ ID NO: 93 (KSGSG), SEQ ID NO: 94(KKSGSG), SEQ ID NO: 95 (EAAAK)2, and SEQ ID NO: 96 (GGAAY). In someembodiments, the nanofiber comprises allergen conjugate peptides and plain self- assembling peptides. In some embodiments, adjacent allergen epitopes in the nanofiber have more than about 0.22 nm, more than about 0.88 nm, about 0.9 nm to about 22 nm, about 10 nm to about 20 nm, more than about 22 nm, less than about 440 nm, more than about 0.22 nm to less than about 440 nm, more than about 0.88 nm to less than about 440 nm, more than about 22 nm to less than about 440 nm, about 25 nm to about 30 nm, or about 28 nm between them. In some embodiments, more than about 0.05%, less than about 1%, less than about 25%, less than about 20%, at least about 0.05%, at least about 0.1%, at least about 0.2%, at least about 0.3%, at least about 0.4%, at least about 0.5%, at least about 0.6%, at least about 0.7%, at least about 0.8%, at least about 0.9%, at leastDocket No.028193-0048-WO01 / 8355 about 1%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% of the peptides in the nanofiber are allergen conjugate peptides. In some embodiments, the nanofiber comprises about 0.2-1% allergen conjugate peptides and 99-99.8% plain self-assembling peptides, or about 0.5% allergen conjugate peptides and 99.5% plain self-assembling peptides. In some embodiments, the nanofiber comprises a plurality of the same or different allergen epitopes. In some embodiments, the nanofiber comprises a combination of 10 to 10,000, or 100 to 10,000 peptides comprising allergen conjugate peptides and plain self-assembling peptides. In some embodiments, the nanofiber comprises a combination of 10 to 10,000, or 100 to 10,000 peptides comprising allergen conjugate peptides, T-cell epitope-conjugate peptides, and plain self-assembling peptides. In some embodiments, the nanofiber comprises a combination of 10 to 10,000, or 100 to 10,000 peptides comprising allergen conjugate peptides, T-cell epitope-conjugate peptides, PAS conjugate peptides or PEG conjugate peptides, and plain self-assembling peptides. In some embodiments, the nanofiber comprises a combination of 10 to 10,000, or 100 to 10,000 peptides comprising allergen conjugate peptides, PAS conjugate peptides or PEG conjugate peptides, and plain self- assembling peptides. In some embodiments, at least about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, or 97.5% of the peptides in the nanofiber are allergen conjugate peptides. In some embodiments, at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, or 15% of the peptides in the nanofiber are T-cell epitope-conjugate peptides. In some embodiments, the allergen conjugate peptide and the T-cell epitope-conjugate peptide are present in the nanofiber at a ratio of about 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 12:1, 14:1, 16:1, 18:1, 20:1, 22:1, 24:1, 26:1, 28:1, 30:1, 32:1, 34:1, 36:1, 38:1, or 40:1. In some embodiments, the self- assembling peptide forms a fibril including beta-sheet structures or a fibril having a coiled coil structure. In some embodiments, the self-assembling peptide forms a fibril having a structure of a helical filament formed around a central axis. In some embodiments, the N- terminus of each self-assembling peptide is positioned at the exterior of the helical filament. In some embodiments, the allergen epitopes are exposed on the exterior surface of the nanofiber. In some embodiments, the nanofiber is about 5-30 nm in width. In some embodiments, the nanofiber is about 100 nm to 1 μm, 100 nm to 2 μm, 100 nm to 3 μm, 100 nm to 4 μm, or 100 nm to 5 μm in length.Docket No.028193-0048-WO01 / 8355

[0009] In another aspect, provided herein is a pharmaceutical composition comprising (a) an allergen conjugate peptide as detailed herein or a nanofiber as detailed herein; and (b) a pharmaceutically acceptable carrier, diluent, and / or excipient. In some embodiments, the pharmaceutical composition further includes (c) an adjuvant selected from cyclic-di-AMP, CpG, cyclic GMP-AMP (cGAMP), cholera toxin B subunit (CTB), retinoic acid, heat labile toxin B subunit, alum, MF59, 3M-052, iscomatrix, squalene-based adjuvants, AS01, AS03, or AS04, or a combination thereof.

[0010] In another aspect, provided herein is a method of treating an allergy. The method may include administering to a subject a therapeutically effective amount of an allergen conjugate peptide as detailed herein, or a nanofiber as detailed herein, or a pharmaceutical composition as detailed herein. In another aspect, provided herein is a method of reducing inflammation in a subject. The method may include administering to a subject a therapeutically effective amount of an allergen conjugate peptide as detailed herein, or a nanofiber as detailed herein, or a pharmaceutical composition as detailed herein. In some embodiments, the allergen conjugate peptide, the nanofiber, or the pharmaceutical composition as administered subcutaneously or sublingually or orally. In some embodiments, the allergen does not leak into the vascular compartment or bloodstream. In some embodiments, the subject forms allergen-specific IgG antibodies. In some embodiments, the allergen-specific IgG antibodies are detected in the subject for at least 20 weeks, at least 21 weeks, at least 22 weeks, at least 23 weeks, at least 24 weeks, at least 25 weeks, at least 30 weeks, at least 35 weeks, at least 40 weeks, at least 45 weeks, at least 50 weeks, at least 51 weeks, or at least 52 weeks. In some embodiments, the subject becomes desensitized to the allergen. In some embodiments, the subject has a reduced Type I hypersensitivity reaction to the allergen. In some embodiments, activation of bone marrow-derived mast cells (BMMC) in the subject is reduced.

[0011] The disclosure provides for other aspects and embodiments that will be apparent in light of the following detailed description and accompanying figures. BRIEF DESCRIPTION OF THE DRAWINGS

[0001] FIG.1 is a diagram showing a peptide-based self-assembling nanofiber platform for use as an allergen vaccine against allergen B cell epitopes that do not elicit systemic anaphylaxis when administered subcutaneously to allergic subjects. In contrast to protein vaccines, the nanofiber vaccines prevent leakage of allergen material into the vascular compartment – a feature that likely underpins their reduced systemic reactogenicity.Docket No.028193-0048-WO01 / 8355 Further, the allergen vaccine platform elicits therapeutic IgG antibody responses capable of desensitizing allergic subjects to allergen-induced Type I hypersensitivity reactions.

[0002] FIG.2 is representative images of a peritoneal mast cell (PMC) gating strategy. PMC were defined as DAPI-CD45+FCER1+CD117+ cells. Activated PMC were defined as CD63+ and reported as a percentage of total PMC.

[0003] FIG.3 is representative images of a BMMC gating strategy. BMMC were defined as DAPI-CD45+FCER1+CD117+ cells. BMMC activation was quantified by CD63 MFI.

[0004] FIGS.4A-4F show that nanofiber allergen vaccines do not elicit systemic anaphylaxis after subcutaneous injection in a mouse model of passive systemic anaphylaxis. FIG.4A is a schematic of Q11 nanofiber allergen vaccine (Nanofiber Vax). FIG.4B is TEM images of (OVA61-68)Q11 nanofibers. Nanofiber formulation: 12.5% (OVA61-68)Q11, 87.5% Q11. FIG.4C is a schematic of model protein-based allergen vaccines (Protein Vax) formed by binding biotin-OVA61-68to streptavidin. FIG.4D is a diagram of a timeline for allergen vaccine challenge experiment. Mice were passively sensitized to the OVA61-68allergen via intravenous injection of 50 g E-C1 IgE mAb and challenged via subcutaneous injection of nanofiber and protein vaccines 24 hours later. Nanofiber formulation: 12.5% (OVA61-68)Q11, 87.5% Q11. 2 mM nanofibers, 100 L per mouse. Allergen tetramer dose: 100 g in 100 L per mouse. FIG.4E is a graph showing body temperature of mice after vaccine challenge. Mixed-effects analysis with Sidak’s multiple comparisons test, * p < 0.05, n = 3 mice. FIG. 4F is a graph showing MCPT1 serum levels before (Pre) and 1 hour after (Post) vaccine challenge. Two-way RM ANOVA with Sidak’s multiple comparisons test, * p < 0.05, n = 3 mice.

[0005] FIGS.5A-5L show that nanofiber and protein vaccines both exhibit allergenicity and activate allergen sensitized mast cells. FIG.5A is a graph showing E-C1 IgE binding curves to nanofiber allergen vaccines (Nanofiber Vax) or Q11 nanofibers alone (Empty Nanofiber). FIG.5B is a graph showing quantification of binding curve AUC for FIG.5A. Unpaired t test, **** p < 0.0001, n = 3 technical replicates. FIG.5C is a graph showing E-C1 IgE binding curves to OVA61-68tetrameric proteins (Protein Vax) and empty tetramers lacking the OVA61-68peptide (Empty Protein). FIG.5D is a graph showing quantification of binding curve AUC for FIG.5C. Unpaired t test, **** p < 0.0001, n = 3 technical replicates. FIG.5E is a graph showing BMMC activation triggered by the nanofiber allergen vaccine as measured by the surface expression of CD63. FIG.5F is a graph showing quantification of maximum BMMC activation for FIG.5E. Unpaired t test, **** p < 0.0001, n = 3 technical replicates. FIG.5G is a graph showing BMMC activation triggered by the protein allergenDocket No.028193-0048-WO01 / 8355 vaccine as measured by the surface expression of CD63. FIG.5H is a graph showing quantification of maximum BMMC activation for FIG.5G. Unpaired t test, **** p < 0.0001, n = 3 technical replicates. FIG.5I is a diagram showing a timeline for peritoneal mast cell activation experiment. To assess peritoneal mast cell activation in response to nanofiber and protein allergen vaccines, mice were passively sensitized to the OVA61-68allergen via intraperitoneal injection of 25 g E-C1 IgE mAb and challenged via intraperitoneal injection of nanofiber and protein vaccines 24 hours later. Mice were sacrificed 1 hour later, and a lavage of the intraperitoneal space was performed. FIG.5J is histograms depicting intraperitoneal mast cell activation quantified by CD63 surface expression. FIG.5K is a graph showing quantification of activated peritoneal mast cell as a percentage of all peritoneal mast cells for nanofiber vaccine. FIG.5L is a graph showing quantification of activated peritoneal mast cell as a percentage of all peritoneal mast cells for protein vaccine. Unpaired t test, * p < 0.05, *** p < 0.001, n = 3 mice.

[0006] FIGS.6A-6I show that nanofiber allergen vaccines prevent leakage of vaccine material into the vascular compartment. FIG.6A is a diagram showing a timeline for systemic vaccine exposure experiment in naïve mice. Fluorescence-matched vaccines labeled with Alexa Fluor®647 (AF647) dye were injected subcutaneously into mice and blood samples were collected over a 24-hour period. FIG.6B is a graph showing serum fluorescence over time. FIG.6C is a graph showing AUC quantification of serum fluorescence over time. Unpaired t test, **** p < 0.0001. FIG.6D is a diagram showing a timeline for systemic vaccine exposure experiment in passively sensitized mice. Mice were passively sensitized to the OVA61-68allergen via intravenous injection of 50 g E-C1 IgE mAb and challenged via subcutaneous injection of fluorescence-matched AF647-labeled nanofiber and protein vaccines 24 hours later. Nanofiber formulation: 6.25% (OVA61-68)Q11, 10% AF647-Q11 and 83.75% Q11. Tetrameric allergen protein (172 g per mouse) contained equimolar OVA61-68dose to the nanofiber vaccine. FIG.6E is a graph showing body temperature of mice after vaccine challenge. Mixed-effects analysis with Sidak’s multiple comparisons test, * p < 0.05, ** p < 0.001. FIG.6F is a graph showing MCPT1 serum levels before (Pre) and 1 hour after (Post) vaccine challenge. Two-way RM ANOVA with Sidak’s multiple comparisons test, **** p < 0.0001. FIG.6G is a graph showing survival curves of mice after vaccine challenge. Endpoint criteria was set at a body temperature of 32°C or lower. Mantel Cox test, * p < 0.05. FIG.6H is a graph showing serum fluorescence over time. FIG.6I is a graph showing AUC quantification of serum fluorescence over time. Unpaired t test, analysis performed on surviving mice only, **** p < 0.0001. n = 5 mice for all experiments.Docket No.028193-0048-WO01 / 8355

[0007] FIGS.7A-7G show that nanofiber allergen vaccines are immunogenic and raise therapeutic sIgG responses that desensitize against allergen challenge in a mouse model of passive systemic anaphylaxis. FIG.7A is a diagram of a timeline of nanofiber allergen vaccine immunization. Mice were primed on Week 0 and boosted on Week 3. Blood samples were collected to assess antibody response. After 6 weeks, mice were sensitized and challenged (see FIGS.7F-G). FIG.7B is graph showing anti-OVA61-68IgG titers over time. FIG.7C is a graph showing anti-OVA61-68IgG titers at Week 4. One-way ANOVA with Dunnett’s multiple comparisons test, **** p < 0.0001, ns p > 0.05, n = 5 mice. FIG.7D is a diagram showing BMMC activation assay workflow. BMMC were sensitized via incubation with E-C1 IgE mAb. 24 hours later, BMMC were exposed to tetrameric OVA61-68model allergen in the presence or absence of purified IgG. FIG.7E is a graph showing BMMC activation that was quantified by CD63 surface expression. One-way ANOVA with Bonferroni multiple comparisons test against the Naïve group, ns p > 0.05, **** p < 0.0001, n = 5 biological replicates. FIG.7F is a diagram showing a timeline for allergen challenge experiment. Mice were passively sensitized to the OVA61-68allergen via intravenous injection of 55 g E-C1 IgE mAb and challenged via intraperitoneal injection of 5 μg tetrameric OVA61-68 model allergen 24 hours later. FIG.7G is a graph showing MCPT1 serum levels before (Pre) and 1 hour after (Post) vaccine challenge. Two-way RM ANOVA with Sidak’s multiple comparisons test, **** p < 0.0001, n = 5 mice.

[0008] FIGS.8A-8I show that nanofiber vaccines against B cell epitope allergen candidates from major peanut allergens raise sIgG responses with therapeutic potential. FIG.8A is a diagram showing a humanized mast cell line (RBL-NFAT-DsRed) activation assay workflow. RBL-NFAT-DsRed were sensitized via incubation with 5% heat-inactivated serum from 7 different peanut allergic human donors. 24 hours later, the cells were exposed to nanofiber vaccines displaying six different peanut allergen epitopes. 16 hours later, mast cell activation was assessed via quantification of DsRed expression. FIG.8B is a graph showing DsRed expression of RBL-NFAT-DsRed cells after stimulation with peanut epitope nanofibers. Crude peanut extract (CE) was included as a positive control to illustrate maximal cell activation. Data is displayed as logarithmic MFI values which, for each donor, are normalized to a Q11-only control. One-way ANOVA with Holm-Sidak multiple comparisons test against the Q11 control group, p-values shown on plot. FIGS.8C-E show immunogenicity of peanut epitope nanofiber vaccines. FIG.8C is graphs showing sIgG titers over time. Mice were primed on Week 0 and boosted on Weeks 3 and 5. sIgG titers assessed against crude peanut extract. FIG.8D is a graph showing week 8 sIgG binding to peanut epitopes. Titers assessed against peanut epitope nanofibers. FIG.8E is a graph showing week 8 sIgG binding to peanut allergen protein. Titers assessed against naturalDocket No.028193-0048-WO01 / 8355 purified or recombinant allergen. FIG.8F is a diagram showing BMMC activation assay workflow. BMMC were sensitized via incubation with peanut allergic serum. 24 hours later, BMMC were exposed to protein peanut allergens in the presence or absence of purified IgG. BMMC activation was quantified by CD63 surface expression. FIGS.8G-8H show that BMMC is sensitized with serum from mice sensitized against crude peanut extract. FIG.8G is a graph showing the effect of AH1a and AH1b sIgG on Ara h 1-induced BMMC activation. AH1b sIgG reduced BMMC activation. FIG.8H is a graph showing the effect of AH3a sIgG on Ara h 3-induced BMMC activation. AH3a sIgG reduced BMMC activation. FIG.8I is a graph showing the effect of AH7a sIgG on Ara h 7-induced BMMC activation. BMMC were sensitized with serum from mice sensitized against recombinant Ara h 7. AH7a sIgG reduced BMMC activation. For FIGS.8G-8I: one-way ANOVA with Dunnett’s multiple comparisons test against the No IgG control group, ns p > 0.05, *** p < 0.001, **** p < 0.0001, n = 3 biological replicates.

[0009] FIGS.9A-9D show nonresponsive BMMC assay results. FIG.9A is a diagram showing an experimental outline of BMMC activation assay. FIGS.9B-9C show the effect of sIgG on BMMC sensitized with serum from crude extract-sensitized mice. FIG.9B is a graph showing that no BMMC activation was detected using Ara h 2. FIG.9C is a graph showing that no BMMC activation was detected using Ara h 7. FIGS.9B-9C indicate inadequate sensitization of mice against these two allergens. FIG.9D is a graph showing the effect of sIgG on BMMC sensitized with serum from Ara h 2-sensitized mice. No BMMC activation was detected using Ara h 2 indicating inadequate sensitization of mice against the allergen.

[0010] FIGS.10A-10B show characterization of IgE responses from mice sensitized to crude peanut extract. FIG.10A is a graph showing IgE responses against protein allergens. FIG.10B is a graph showing IgE responses against peptide epitopes displayed on Q11 nanofibers.

[0011] FIG.11 is a diagram showing a mode of hypoallergenic immunogen design that enables immunization against IgE-reactive peptide B cell epitopes by optimizing the distance between epitopes. Using DNA-based model immunogens, mast cells and B cells exhibit idiosyncratic sensitivity to inter-epitope spacing, with mast cell activation being dampened by high inter-epitope spacing while B cells remain responsive to identical immunogen configurations.

[0012] FIGS.12A-12G show an overview of a hypothesized strategy for hypoallergenic immunogen design project workflow schematic. FIG.12A is a schematic showing theDocket No.028193-0048-WO01 / 8355 differential impact of inter-epitope spacing on cellular activation in mast cells and B cells based on reports from the literature. Recent evidence suggests that mast cell activation is decreased in response to increasing space between IgE-reactive epitopes whereas B cell activation through the BCR may remain sensitive to similar immunogen configurations. FIG. 12B is a diagram showing a description of the general project workflow. First, the existence of the hypothesized hypoallergenic zone (shown in FIG.12A) was validated using DNA SST model immunogens with precise control over nanoscale spatial parameters to elucidate the impact of immunogen valency and inter-epitope spacing on mast cell and B cell activation. Motivated by these findings, hypoallergenic supramolecular peptide nanofiber immunogens were designed that displayed IgE-reactive peptide epitopes at ultra-low densities and their use as allergen vaccine immunogens was explored in a mouse model of allergy. FIG.12C is schematics of 6-helix-bundle SST structure. Left: Cross-section of 6-helix-bundle as viewed in oxView.60. Center: Strand diagram showing a short section of six-helix-bundle. A single- stranded overhang is incorporated for binding to DNP modified DNA oligo. Right: 3D models illustrating the formulation of a 4xDNP structure. FIG.12D is AFM images of 2-streptavidin within 28 nm spacings on 6-helix-bundle SST structures. FIG.12E is AFM images of 3- streptavidin within 28 nm spacings on 6-helix-bundle SST structures. FIG.12F is AFM images of 4-streptavidin within 28 nm spacings on 6-helix-bundle SST structures. The scale bars for the insets are 200 nm. FIG.12G is an agarose gel image of 6-helix bundle without or with 4x DNP spaced apart by 7, 14, 21, 28 nm. M: DNA marker.

[0013] FIGS.13A13-C show an SST model immunogen design. FIG.13A is diagram showing CaDNAno design of the 6-helix-bundle DNA SST structure. FIG.13B is a diagram showing models of the 6-helix-bundle as viewed in oxView (cross-section and side view). FIG.13C is schematics of 6-helix-bundle structures with four DNP placed at different spacing of 7 nm, 14 nm, 21 nm, and 28 nm.

[0014] FIGS.14A-14L show that valency and inter-epitope spacing are majordeterminants of mast cell activation through IgE-Fc RI and multivalent model immunogenswith high inter-epitope spacing attenuate mast cell activation in vitro and in vivo. FIG.14A is a diagram showing a BMMC assay workflow schematic. BMMC were sensitized with monoclonal mouse anti-DNP IgE (SPE-7) and stimulated with DNP SST model immunogens. BMMC degranulation was quantified via CD63 surface expression. FIG.14B is a graph showing average BMMC activation quantification for varying DNP concentrations for bivalent 2xDNP SST configurations using Mean Fluorescent Intensity (MFI). FIG.14C is a graph showing maximum activation for the data of FIG.14B. FIG.14D is a graph showing average BMMC activation quantification for varying DNP concentrations for trivalent 3xDNPDocket No.028193-0048-WO01 / 8355 SST configurations using MFI. FIG.14E is a graph showing maximum activation for the data of FIG.14D. FIG.14F is a graph showing average BMMC activation quantification for varying DNP concentrations for tetravalent 4xDNP SST configurations using MFI. FIG.14G is a graph showing maximum activation for the data of FIG.14F. For FIGS.14B-14G, all data is normalized to a DNP-BSA positive control (set to 1.0) and a 6HB (no DNP SST) negative control (set to 0.0). FIG.14H is a graph showing percentage of cells activated by 4xDNP SST structures with various inter-epitope spacings. FIG.14I is a graph showing CD63 expression of cells activated by 4xDNP SST structures with various inter-epitope spacings. FIG.14J is a diagram showing a PMC activation assay workflow schematic. Mice were sensitized with monoclonal mouse anti-DNP IgE (SPE-7) and subsequently challenged with 4xDNP SST model immunogens intraperitoneally. PMC were retrieved via peritoneal lavage and activation was quantified via CD63 surface expression. FIG.14K is a representative diagram showing CD63 surface expression. FIG.14L is a graph showing PMC activation quantification varying DNP concentrations using MFI. For FIGS.14B-14I, n=3 technical replicates. Statistical significance was determined using a one-way ANOVA with Tukey’s multiple comparisons test (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns = not significant). For FIGS.14J-14L, n=3 biological replicates. Statistical significance determined using a one-way ANOVA with Dunnett’s multiple comparisons test to the 6HB group (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns = not significant). All p values are provided in TABLE 5. All error bars represent SD; dotted lines represent means.

[0015] FIGS.15A-15B show 4xDNP SST model immunogen IgE binding ELISA and correlation with BMMC activation. FIG.15A is a graph showing SPE-7 IgE binding. 4xDNP SST model immunogens were coated onto ELISA plate wells and SPE-7 IgE binding was assessed. FIG.15B is a graph showing a correlation for IgE binding curve AUC values with BMMC activation. Pearson correlation coefficient, r, is shown; ns, p > 0.05.

[0016] FIGS.16A-16D show a RAMOSDNPcell design overview and validation. FIG. 16A is a schematic of the chimeric DNP-specific BCR constructs. The heavy chain variable region of SPE-7 IgE (SPE-7 VH) was attached to the constant region of human membrane- bound IgM (mIGHM CH). The light chain variable region of SPE-7 IgE (SPE-7 VL) was attached to the constant region of human lambda light chain (IGL1 CL). BCR negative cells were sorted out of the parental RAMOS cell line and the chimeric BCR genes were inserted using a PiggyBac transposase system. BCR expressing cells were sorted out and expanded to generate the RAMOSDNPcell line. FIG.16B is a graph showing IgM BCR expression for BCR negative RAMOS cells (dotted line, left) and after anti-DNP BCR gene insertion (solid gray, right). FIGS.16C-16D show functional validation of RAMOSDNPcells using calcium fluxDocket No.028193-0048-WO01 / 8355 assay. FIG.16C is a graph showing calcium flux for BCR negative RAMOS cells before BCR gene insertion that were stimulated with the calcium ionophore ionomycin as a positive control and DNP-BSA at 10 ng / mL. FIG.16D is a graph showing calcium flux for RAMOSDNP cells that were stimulated with the calcium ionophore ionomycin as a positive control and DNP-BSA at 10 ng / mL. As expected, only RAMOSDNPcells were responsive to DNP-BSA stimulation.

[0017] FIGS.17A-17K show that B cell activation through the BCR remains responsive to multivalent immunogens with high inter-epitope spacing. FIG.17A is a diagram showing B cell activation (pERK) assay workflow schematic. RAMOSDNPcells were stimulated with DNP-BSA or DNP SST immunogens with various inter-epitope spacings at 20 nM DNP. B cell activation was quantified via pERK expression measured by flow cytometry. FIG.17B is a graph showing pERK expression in unstimulated cells or cells stimulated with immunogens for 1 and 5 minutes. FIG.17C is representative histograms from the 5-minute timepoint of FIG.17B. FIG.17D is a graph showing a comparison of mast cell activation and B cell activation elicited by 4xDNP SST model immunogens. B cell activation data corresponds to the 5-minute timepoint of pERK expression. Mast cell activation data corresponds to the maximum CD63 expression elicited by each immunogen in BMMC. Black line indicates the ratio of B cell to mast cell activation as displayed on the right y-axis. FIG.17E is a diagram showing B cell calcium flux assay workflow schematic. RAMOSDNPcells were labeled with Fluo-4 intracellular calcium indicator and stimulated with DNP-BSA or SST immunogens with various inter-epitope spacings at 20 nM DNP. FIG.17F is a graph showing calcium flux measured via flow cytometry for bivalent 2xDNP SST structures. FIG.17G is a graph showing calcium flux measured via flow cytometry for trivalent 3xDNP SST structures. FIG. 17H is a graph showing calcium flux measured via flow cytometry for tetravalent 4xDNP SST structures. Area under curve (AUC) for each is displayed on the bottom. FIG.17I is a graph showing distribution of Fluo-4 calcium probe signal in cells at baseline (grey) and during the peak of the flux curve after stimulation (colored). FIG.17J is a graph showing the percentage of cells that exhibit calcium signal above baseline. FIG.17K is a graph showing the Fluo-4 MFI of those activated cells. For both the pERK and calcium flux assays, data is normalized to the DNP-BSA positive control (set to 1.0) and the 6HB (no DNP SST) negative control (set to 0.0). For FIG.17B, statistical significance was determined using a two-way ANOVA with Tukey’s multiple comparisons test (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns = not significant). n = 5 for unstimulated and 5-minute timepoint (two experimental repeats with n = 2 and n = 3). n = 3 for 1-minute timepoint. For all other analyses, statistical significance was determined using a one-way ANOVA with Tukey’s multiple comparisons test (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns = notDocket No.028193-0048-WO01 / 8355 significant). n = 3 technical replicates. All p values are provided in TABLE 6. All error bars represent SD; dotted lines represent mean.

[0018] FIGS.18A-18H show that the Coil29 supramolecular vaccine platform enables tunable stoichiometric control over immunogen epitope density. FIG.18A is a schematic of the approach to control epitope density on C29 nanofibers by varying the stoichiometric ratio of (Epitope)C29 to C29 peptide within the nanofiber formulation. Examples of high and low density formulation with low and high inter-epitope spacing, respectively, are shown. FIGS. 18B-18G are images and graphs showing confirmation of the ability to co-assemble multiple peptide species into uniform nanofibers, (TAMRA-OVA)C29 and (FITC-OVA)C29 were co- assembled into C29 nanofibers alone (FIGS.18B-18C) or together (FIG.18D), each at 10% of the total peptide for all formulations. FIG.18B is images showing co-assembly of (TAMRA-OVA)C29. FIG.18C is images showing co-assembly of (FITC-OVA)C29. FIG. 18D is images showing co-assembly of (TAMRA-OVA FITC-OVA)C29. The large images display merged signal from FITC, TAMRA, and FRET channels (FRET channel = excitation of FITC and emission of TAMRA). Each image channel is adjusted for brightness and contrast equally for all formulations. FIG.18E is a graph showing quantification of average pixel intensity of individual fibers for (TAMRA-OVA)C29. FIG.18F is a graph showing quantification of average pixel intensity of individual fibers for (FITC-OVA)C29. FIG.18G is a graph showing quantification of average pixel intensity of individual fibers for (TAMRA- OVA FITC-OVA)C29. OVA indicates the OVA323-339peptide. Subscript denotes percentage of peptide species. A decrease in the donor (FITC) signal and an increase in the FRET signal was observed in nanofibers with co-assembled fluorophores, indicating co-assembly of peptide species. Pixel intensity values were compensated by subtracting the crosstalk of each fluorophore into the two other channels using the single-fluorophore formulations. FIG. 18H is a graph showing frequency of C29 nanofiber immunogens co-assembled with varying ratios of TAMRA-labeled (OVA)C29 imaged on a confocal microscope. TAMRA signal of nanofibers co-assembled with 10%, 2.5%, and 1.25% TAMRA-OVA-C29 were quantified and binned based on MPI. The relative frequency of fibers within each bin as a percentage of total fibers is shown. MPI, Mean Pixel Intensity. For FIGS.18E-18G, statistical significance was determined using a one-way ANOVA with Tukey’s multiple comparisons test (****p < 0.0001). All p values are provided in TABLE 7. Dotted lines represent mean.

[0019] FIG.19 is TAMRA-OVA)C29 nanofiber titration confocal images and ROI masks to identify individual nanofibers as shown in FIG.18H.

[0020] FIGS.20A-20B show a theoretical relationship between epitope stoichiometry and inter-epitope spacing along the coil29 nanofiber axis. FIG.20A is a conceptualDocket No.028193-0048-WO01 / 8355 schematic showing how inter-epitope spacing depends on epitope stoichiometry. FIG.20B is a graph showing a relationship between epitope stichometry and the theoretical distance between epitopes assuming perfectly uniform distribution into the Coil29 nanofiber scaffold. Inter-epitope spacing is calculated as the distance between epitopes along the fiber axis and does not account for epitope orientation.

[0021] FIGS.21A-21I show that Coil29 allergen vaccine immunogens with ultra-low epitope density activate B cells while avoiding mast cell activation. FIG.21A is a diagram showing a BMMC assay workflow schematic. BMMC were sensitized with monoclonal mouse anti-allergen IgE and later stimulated with nanofiber immunogens at various epitope densities. FIG.21B is a graph showing BMMC activation via MFI that was assessed at different nanofiber concentrations for BMMC that were sensitized with anti-DNP IgE (SPE-7) and stimulated with (DNP)C29 nanofibers. FIG.21C is a graph showing maximum BMMC activation for the data in FIG.21B. FIG.21D is a graph showing BMMC activation via MFI that was assessed at different nanofiber concentrations for BMMC that were sensitized with anti-OVA61-68IgE and stimulated with (OVA61-68)C29 nanofibers. FIG.21E is a graph showing maximum BMMC activation for the data in FIG.21D. For FIGS.21B-21E, the percentage refers to the ratio of (Epitope)C29 relative to C29. FIG.21F is a diagram showing a B cell activation (pERK) assay workflow schematic. RAMOSDNPcells were stimulated with high and ultra-low density (DNP)C29 nanofibers for 5 minutes at 100 nM DNP. Downstream ERK phosphorylation was quantified via flow cytometry (FIGS.21G-21I). FIG.21G is a representative histogram of pERK expression. FIG.21H is a representative histogram of activated B cells as a percent of total cells. FIG.21I is a representative histogram of the pERK MFI of activated cells. The dotted line in FIG.21I indicates the pERK MFI of unstimulated cells. For FIGS.21G-21I, the subscript denotes the percentage of (Epitope)C29 relative to C29. n=3 technical replicates. Statistical significance was determined using a one-way ANOVA with Tukey’s multiple comparisons test (****p < 0.0001, ns = not significant). All p values are provided in TABLE 8. All error bars represent SD; dotted lines represent mean.

[0022] FIGS.22A-22J show that ultra-low density hypoallergenic Coil29 immunogens elicit therapeutic allergen-specific IgG responses that neutralize allergen and protect mice from allergen challenge in a model of passive sensitization and anaphylaxis. FIG.22A is a diagram showing a nanofiber allergen vaccine immunogenicity and allergen challenge experimental workflow schematic. First, mice were sensitized to the allergen OVA61-68via weekly intraperitoneal injections of KLH-OVA61-68 and AlHydrogel. Splenocytes from OVA61-68sensitized mice were adoptively transferred to recipient mice which were then immunizedDocket No.028193-0048-WO01 / 8355 with nanofiber vaccines at weeks 0, 3, and 5. On week 6, mice were passively sensitized via intravenous injection of anti-OVA61-68IgE (E-C1) and then challenged with a model allergen tetramer formed by loading streptavidin with biotinylated OVA61-68 peptide. Body temperature and spikes in serum MCPT-1 levels were monitored as indicators of systemic anaphylaxis. FIG.22B is graphs showing anti-OVA61-68IgG responses in serum of mice immunized with the indicated nanofiber vaccine formulation. FIG.22C is a graph showing titers at week 5 post-prime. n = 5 biological replicates. Statistical significance was determined using a one-way ANOVA with Dunnett’s multiple comparisons test against the Naïve group (**p < 0.01, ***p < 0.001, ****p < 0.0001, ns = not significant). FIG.22D is a graph showing results from a BMMC activation assay of IgG purified from the pooled serum of immunized mice was tested for allergen neutralization. n = 3 technical replicates using IgG purified from pooled serum from each group. Statistical significance was determined using a one-way ANOVA with Tukey’s multiple comparisons test (***p < 0.001, ****p < 0.0001, ns = not significant). FIG.22E is a graph showing body temperature of passively sensitized mice after allergen challenge. FIG.22F is a graph showing the change in body temperature from baseline. n = 5 biological replicates. Statistical significance was determined using a one-way ANOVA with Dunnett’s multiple comparisons test against the Naïve group (****p < 0.0001). FIG.22G is a graph showing MCPT-1 levels in the serum of mice immunized with the 25% nanofiber vaccine formulation before (Pre) and 1 hour after (Post) allergen challenge. FIG.22H is a graph showing MCPT-1 levels in the serum of mice immunized with the 0.5% nanofiber vaccine formulation before (Pre) and 1 hour after (Post) allergen challenge. FIG.22I is a graph showing MCPT-1 levels in the serum of naïve mice before (Pre) and 1 hour after (Post) allergen challenge. FIG.22J is a graph showing the change in MCPT-1 levels from baseline. n = 5 biological replicates. Statistical significance was determined using a one-way ANOVA with Dunnett’s multiple comparisons test against the Naïve group (**p < 0.01, ***p < 0.001). All p values are provided in TABLE 9. All error bars represent SD; dotted lines represent mean.

[0023] FIG.23 is AFM images of (OVA61-68)C29 nanofibers used for immunization.

[0024] FIG.24 is a graph showing week 3 anti-Coil29 scaffold IgG titers from (OVA61-68)C29 immunized mice. Serum was collected from the immunizations outlined in FIGS. 22A-22J at week 3 post-prime and analyzed for anti-C29 IgG via ELISA. Statistical significance was determined using a one-way ANOVA with Dunnett’s multiple comparisons test against the (OVA61-68)25C29 immunization group (*p < 0.05, ***p < 0.001).Docket No.028193-0048-WO01 / 8355 DETAILED DESCRIPTION

[0012] Provided herein are novel peptide conjugates that self-assemble into peptide nanofibers or fibrils, with an allergen epitope conjugated thereto. These peptide conjugates may be used to treat allergies.

[0013] As detailed herein, the inventors used a supramolecular nanofiber vaccine system as an allergen vaccine platform. Because of the safety concerns associated with allergen vaccination, the allergic responses to the vaccine were studied. It was demonstrated that in contrast to protein-based allergen vaccines, the nanofiber platform does not induce systemic allergic responses after subcutaneous injection. Mechanistically, unlike protein vaccines, nanofiber vaccines may prevent leakage of allergen material into the vascular compartment, a feature that likely underpins their reduced systemic reactogenicity. Further, the allergen vaccine platform elicits therapeutic IgG antibody responses capable of desensitizing allergic mice to allergen-induced Type I hypersensitivity reactions, and the platform can raise therapeutic sIgG responses to broadly reactive epitopes from, for example, major peanut allergens. A nanofiber allergen vaccine platform as detailed herein may be effective in achieving allergen desensitization while limiting vaccine-related adverse events.

[0014] As further detailed herein, the new mode of hypoallergenic allergen vaccine immunogen design allows for immunization against IgE-reactive B cell epitopes. Mast cells and B cells respond to multivalent antigens differently (FIG.4A). Allergen-induced mast cellsignaling through Fc RI-IgE is affected by the distance that separates IgE-reactive epitopes,with more distant inter-epitope spacing yielding lower mast cell activation. Like mast cellactivation though Fc RI, B cell activation through the BCR benefits from cross-linking bymultivalent antigen. Also similar to mast cells, B cell activation is enhanced by increasing antigen valency. However, despite the similarities between the activation mechanisms of the two cell types, in contrast to mast cells, B cell activation is maintained at distant inter-epitope spacings. Given this idiosyncratic sensitivity to epitope spacing between mast cells and B cells, the inventors discovered that immunogens bearing IgE-reactive B cell epitopes may be made hypoallergenic if the epitopes can be spaced far enough away from each other. Single-stranded tile (SST) DNA nanostructures were used to construct model immunogens and design a new class of hypoallergenic allergen vaccine immunogen based on supramolecular peptide nanofibers with ultra-low epitope density (FIG.4B). DNA-based structures enabled the arrangement and spacing of features on nanoscale objects for the studies detailed herein, however, their brief stability in vivo limits their use as vaccines. DNA SST structures were used to investigate the epitope spacing responses of mast cells and BDocket No.028193-0048-WO01 / 8355 cells in vitro, and these findings were applied to the design of supramolecular peptide immunogens possessing the in vivo stability for translation to clinical use. Mast cell activation can be dampened by high inter-epitope spacing while B cells remain responsive to identical immunogen configurations. Controlling the distances between IgE-reactive epitopes may enable allergen vaccination against IgE-reactive epitope targets in the absence of allergic reactogenicity. The supramolecular peptide nanofibers with ultra-low epitope density can thereby be used as an allergen vaccine and raise protective allergen- neutralizing IgG antibody responses. 1. Definitions

[0015] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In case of conflict, the present document, including definitions, will control. Preferred methods and materials are described below, although methods and materials similar or equivalent to those described herein can be used in practice or testing of the present invention. All publications, patent applications, patents and other references mentioned herein are incorporated by reference in their entirety. The materials, methods, and examples disclosed herein are illustrative only and not intended to be limiting.

[0016] The terms “comprise(s),” “include(s),” “having,” “has,” “can,” “contain(s),” and variants thereof, as used herein, are intended to be open-ended transitional phrases, terms, or words that do not preclude the possibility of additional acts or structures. The singular forms “a,” “and” and “the” include plural references unless the context clearly dictates otherwise. The present disclosure also contemplates other embodiments “comprising,” “consisting of” and “consisting essentially of,” the embodiments or elements presented herein, whether explicitly set forth or not.

[0017] For the recitation of numeric ranges herein, each intervening number there between with the same degree of precision is explicitly contemplated. For example, for the range of 6-9, the numbers 7 and 8 are contemplated in addition to 6 and 9, and for the range 6.0-7.0, the number 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, and 7.0 are explicitly contemplated.

[0018] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In case of conflict, the present document, including definitions, will control. Preferred methods and materials are described below, although methods and materials similar or equivalent toDocket No.028193-0048-WO01 / 8355 those described herein can be used in practice or testing of the present invention. All publications, patent applications, patents and other references mentioned herein are incorporated by reference in their entirety. The materials, methods, and examples disclosed herein are illustrative only and not intended to be limiting.

[0019] The term “about” or “approximately” as used herein as applied to one or more values of interest, refers to a value that is similar to a stated reference value, or within an acceptable error range for the particular value as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined, such as the limitations of the measurement system. In certain aspects, the term “about” refers to a range of values that fall within 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in either direction (greater than or less than) of the stated reference value unless otherwise stated or otherwise evident from the context (except where such number would exceed 100% of a possible value). Alternatively, “about” can mean within 3 or more than 3 standard deviations, per the practice in the art. Alternatively, such as with respect to biological systems or processes, the term “about” can mean within an order of magnitude, preferably within 5-fold, and more preferably within 2- fold, of a value.

[0020] The term “adjuvant” refers to a compound or mixture that enhances the immune response to an antigen. Adjuvants may contain a substance to protect the antigen from rapid catabolism, such as aluminum hydroxide or a mineral oil, and also a protein derived from lipid A, Bortadella pertussis, or Mycobacterium tuberculosis. Suitable adjuvants may be commercially available and include, for example, complete or incomplete Freund's adjuvant; AS-2; aluminum salts such as aluminum hydroxide (as a gel, where appropriate) or aluminum phosphate; alum; MF59; calcium salts, iron salts, or zinc salts; an insoluble suspension of acylated tyrosine; acylated sugars; cationically or anionically derivatized polysaccharides; polyphosphazenes; biologically degradable microspheres; monophosphoryl lipid A, cytokines such as GM-CSF, Interleukin-2, Interleukin-7, Interleukin-12, CpG, cholera toxin B subunit (CTB), retinoic acid, heat labile toxin B subunit, 3M-052, iscomatrix, squalene-based adjuvants, AS01, AS03, AS04, STING agonists like cyclic dinucleotides, cyclic-di-AMP, CpG, cyclic GMP-AMP (cGAMP), or a combination thereof.

[0021] “Amino acid” as used herein refers to naturally occurring and non-natural synthetic amino acids, as well as amino acid analogs and amino acid mimetics that function in a manner similar to the naturally occurring amino acids. Naturally occurring amino acids are those encoded by the genetic code. Amino acids can be referred to herein by either their commonly known three-letter symbols or by the one-letter symbols recommended byDocket No.028193-0048-WO01 / 8355 the IUPAC-IUB Biochemical Nomenclature Commission. Amino acids include the side chain and polypeptide backbone portions.

[0022] As used herein, the term “antigen” is a molecule capable of being bound by an antibody or B-cell receptor or T-cell receptor. The term “antigen” encompasses B-cell epitopes. The term “antigen” may also encompass T-cell epitopes. An antigen also refers to a molecule against which a subject can initiate a humoral and / or cellular immune response leading to the activation of B-lymphocytes and / or T-lymphocytes. An antigen is capable of inducing a humoral immune response and / or cellular immune response leading to the production of B- and / or T-lymphocytes. The structural aspect of an antigen that gives rise to a biological response is referred to herein as an “antigenic determinant.” B-lymphocytes respond to foreign antigenic determinants via antibody production, whereas T-lymphocytes are the mediator of cellular immunity. Thus, antigenic determinants or epitopes are those parts of an antigen that are recognized by antibodies, or in the context of an MHC, by T-cell receptors. An antigenic determinant need not be a contiguous sequence or segment of protein and may include various sequences that are not immediately adjacent to one another. In some embodiments, the antigen contains or is linked to a Th cell epitope. An antigen can have one or more epitopes (B-epitopes and T-epitopes). Antigens may also be mixtures of several individual antigens. Antigens can be any type of biologic molecule including, for example, simple intermediary metabolites, sugars, lipids, and hormones as well as macromolecules such as complex carbohydrates, phospholipids, nucleic acids and proteins. Common categories of antigens include, but are not limited to, viral antigens, bacterial antigens, fungal antigens, protozoa and other parasitic antigens, tumor antigens, antigens involved in autoimmune disease, allergy and graft rejection, and other miscellaneous antigens. Antigens can be microbial antigens, such as viral, fungal, or bacterial; or therapeutic antigens such as antigens associated with cancerous cells or growths, or autoimmune disorders. In some embodiments, the antigen is selected from a small molecule, nucleotide, polynucleotide, peptide, polypeptide, protein, lipid, carbohydrate, other immunogenic molecules, and a combination thereof. In some embodiments, the antigen is an allergen. An allergen is a substance that can cause an allergic reaction in a subject.

[0023] The terms “control,” “reference level,” and “reference” are used herein interchangeably. The reference level may be a predetermined value or range, which is employed as a benchmark against which to assess the measured result. “Control group” as used herein refers to a group of control subjects. The predetermined level may be a cutoff value from a control group. The predetermined level may be an average from a controlDocket No.028193-0048-WO01 / 8355 group. Cutoff values (or predetermined cutoff values) may be determined by Adaptive Index Model (AIM) methodology. Cutoff values (or predetermined cutoff values) may be determined by a receiver operating curve (ROC) analysis from biological samples of the patient group. ROC analysis, as generally known in the biological arts, is a determination of the ability of a test to discriminate one condition from another, e.g., to determine the performance of each marker in identifying a patient having CRC. A description of ROC analysis is provided in P.J. Heagerty et al. (Biometrics 2000, 56, 337-44), the disclosure of which is hereby incorporated by reference in its entirety. Alternatively, cutoff values may be determined by a quartile analysis of biological samples of a patient group. For example, a cutoff value may be determined by selecting a value that corresponds to any value in the 25th-75th percentile range, preferably a value that corresponds to the 25th percentile, the 50th percentile or the 75th percentile, and more preferably the 75th percentile. Such statistical analyses may be performed using any method known in the art and can be implemented through any number of commercially available software packages (e.g., from Analyse-it Software Ltd., Leeds, UK; StataCorp LP, College Station, TX; SAS Institute Inc., Cary, NC.). The healthy or normal levels or ranges for a target or for a protein activity may be defined in accordance with standard practice. A control may be a subject or cell without a composition as detailed herein. A control may be a subject, or a sample therefrom, whose disease state or infection is known. The subject, or sample therefrom, may be healthy, diseased or infected, diseased or infected prior to treatment, diseased or infected during treatment, or diseased or infected after treatment, or a combination thereof.

[0024] “Identical” or “identity” as a percentage as used herein in the context of two or more polynucleotide or polypeptide sequences means that the sequences have a specified percentage of residues that are the same over a specified region. The percentage may be calculated by optimally aligning the two sequences, comparing the two sequences over the specified region, determining the number of positions at which the identical residue occurs in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the specified region, and multiplying the result by 100 to yield the percentage of sequence identity. In cases where the two sequences are of different lengths or the alignment produces one or more staggered ends and the specified region of comparison includes only a single sequence, the residues of single sequence are included in the denominator but not the numerator of the calculation. When comparing DNA and RNA, thymine (T) and uracil (U) may be considered equivalent. Identity may be performed manually or by using a computer sequence algorithm such as BLAST or BLAST 2.0.Docket No.028193-0048-WO01 / 8355

[0025] “Immunogenicity” refers to the ability of an antigen to induce an immune response and includes the intrinsic ability of an antigen to generate antibodies in a subject. In some embodiments, the self-assembling peptides described herein, or the nanofibers they form, are not immunogenic without an antigen appended thereto.

[0026] “Nucleic acid” or “oligonucleotide” or “polynucleotide” as used herein means at least two nucleotides covalently linked together. The depiction of a single strand also defines the sequence of the complementary strand. Thus, a polynucleotide also encompasses the complementary strand of a depicted single strand. Many variants of a polynucleotide may be used for the same purpose as a given polynucleotide. Thus, a polynucleotide also encompasses substantially identical polynucleotides and complements thereof. A single strand provides a probe that may hybridize to a target sequence under stringent hybridization conditions. Thus, a polynucleotide also encompasses a probe that hybridizes under stringent hybridization conditions. Polynucleotides may be single stranded or double stranded or may contain portions of both double stranded and single stranded sequence. The polynucleotide can be nucleic acid, natural or synthetic, DNA, genomic DNA, cDNA, RNA, mRNA, or a hybrid, where the polynucleotide can contain combinations of deoxyribo- and ribo-nucleotides, and combinations of bases including, for example, uracil, adenine, thymine, cytosine, guanine, inosine, xanthine hypoxanthine, isocytosine, and isoguanine. Polynucleotides can be obtained by chemical synthesis methods or by recombinant methods.

[0027] A “peptide” or “polypeptide” is a linked sequence of two or more amino acids linked by peptide bonds. The polypeptide can be natural, synthetic, or a modification or combination of natural and synthetic. Peptides and polypeptides include proteins such as binding proteins, receptors, and antibodies. The terms “polypeptide”, “protein,” and “peptide” are used interchangeably herein. “Primary structure” refers to the amino acid sequence of a particular peptide. “Secondary structure” refers to locally ordered, three dimensional structures within a polypeptide. Secondary structure may include beta-sheet and alpha- helices. These structures are commonly known as domains, e.g., enzymatic domains, extracellular domains, transmembrane domains, pore domains, and cytoplasmic tail domains. Domains are portions of a polypeptide that form a compact unit of the polypeptide and are typically 15 to 350 amino acids long. Exemplary domains include domains with enzymatic activity or ligand binding activity. Typical domains are made up of sections of lesser organization such as stretches of beta-sheet and alpha-helices. “Tertiary structure” refers to the complete three dimensional structure of a polypeptide monomer. “Quaternary structure” refers to the three dimensional structure formed by the noncovalent association ofDocket No.028193-0048-WO01 / 8355 independent tertiary units. A “motif” is a portion of a polypeptide sequence and includes at least two amino acids. A motif may be 2 to 20, 2 to 15, or 2 to 10 amino acids in length. In some embodiments, a motif includes 3, 4, 5, 6, or 7 sequential amino acids. A domain may be comprised of a series of the same type of motif.

[0028] The phrases “pharmaceutically acceptable” or “pharmacologically acceptable” refer to molecular entities and compositions that do not produce an adverse, allergic, or other untoward reaction when administered to an animal, or human. As used herein, “pharmaceutically acceptable carrier” includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like. In some embodiments, a carrier includes a solution at neutral pH. In some embodiments, a carrier includes a salt. In some embodiments, a carrier includes a buffered solution. In some embodiments, a carrier includes phosphate buffered saline solution.

[0029] “Sample” or “test sample” as used herein can mean any sample in which the presence and / or level of a target is to be detected or determined or a portion from a subject or portion of a composition as detailed herein. Samples may include liquids, solutions, emulsions, or suspensions. Samples may include a medical sample. Samples may include any biological fluid or tissue, such as blood, whole blood, fractions of blood such as plasma and serum, muscle, interstitial fluid, sweat, saliva, urine, tears, synovial fluid, bone marrow, cerebrospinal fluid, nasal secretions, sputum, amniotic fluid, bronchoalveolar lavage fluid, gastric lavage, emesis, fecal matter, lung tissue, peripheral blood mononuclear cells, total white blood cells, lymph node cells, spleen cells, tonsil cells, cancer cells, tumor cells, bile, digestive fluid, skin, or combinations thereof. In some embodiments, the sample comprises an aliquot. In other embodiments, the sample comprises a biological fluid. Samples can be obtained by any means known in the art. The sample can be used directly as obtained from a patient or can be pre-treated, such as by filtration, distillation, extraction, concentration, centrifugation, inactivation of interfering components, addition of reagents, and the like, to modify the character of the sample in some manner as discussed herein or otherwise as is known in the art.

[0030] “Subject” and “patient” as used herein interchangeably refers to any vertebrate, including, but not limited to, a mammal that wants or is in need of the herein described compositions or methods. The subject may be a human or a non-human. The subject may be a vertebrate. The subject may be a mammal. The mammal may be a primate or a non- primate. The mammal can be a non-primate such as, for example, cow, pig, camel, llama, hedgehog, anteater, platypus, elephant, alpaca, horse, goat, rabbit, sheep, hamster, guinea pig, cat, dog, rat, and mouse. The mammal can be a primate such as a human. TheDocket No.028193-0048-WO01 / 8355 mammal can be a non-human primate such as, for example, monkey, cynomolgous monkey, rhesus monkey, chimpanzee, gorilla, orangutan, and gibbon. The subject may be of any age or stage of development, such as, for example, an adult, an adolescent, a child, such as age 0-2, 2-4, 2-6, or 6-12 years, or an infant, such as age 0-1 years. The subject may be male. The subject may be female. In some embodiments, the subject has a specific genetic marker. The subject may be undergoing other forms of treatment.

[0031] “Substantially identical” can mean that a first and second amino acid or polynucleotide sequence are at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%, or less than 100% over a region of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1100 amino acids or nucleotides, respectively.

[0032] “Treatment” or “treating” or “therapy” when referring to protection of a subject from a disease, means suppressing, repressing, reversing, alleviating, ameliorating, or inhibiting the progress of disease, or completely eliminating a disease. A treatment may be either performed in an acute or chronic way. The term also refers to reducing the severity of a disease or symptoms associated with such disease prior to affliction with the disease. Treatment may result in a reduction in the incidence, frequency, severity, and / or duration of symptoms of the disease. Preventing the disease involves administering a composition of the present invention to a subject prior to onset of the disease. Suppressing the disease involves administering a composition of the present invention to a subject after induction of the disease but before its clinical appearance. Repressing or ameliorating the disease involves administering a composition of the present invention to a subject after clinical appearance of the disease. A disease may include a bacterial infection. Bacterial infections may occur when bacteria enter the body of a subject, increase in number, and cause a reaction in the body. Bacteria can enter the body through an opening in the skin of a subject, such as a cut or surgical wound, or through orifices in the body of a subject, such as the airway (for example, nasal passages, mouth), urethra, ear canal, eye, and the like. A bacterial infection can be caused by either gram-negative or gram-positive bacteria. In some embodiments, the disease includes inflammation. In some embodiments, the disease includes sepsis. In some embodiments, the disease includes an allergy.

[0033] “Variant” used herein with respect to a polynucleotide means (i) a portion or fragment of a referenced nucleotide sequence; (ii) the complement of a referenced nucleotide sequence or portion thereof; (iii) a nucleic acid that is substantially identical to a referenced nucleic acid or the complement thereof; or (iv) a nucleic acid that hybridizesDocket No.028193-0048-WO01 / 8355 under stringent conditions to the referenced nucleic acid, complement thereof, or a sequence substantially identical thereto. A variant can be a polynucleotide sequence that is substantially identical over the full length of the full polynucleotide sequence or a fragment thereof. The polynucleotide sequence can be 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, or less than 100% identical over the full length of the polynucleotide sequence or a fragment thereof.

[0034] “Variant” with respect to a peptide or polypeptide that differs in amino acid sequence by the insertion, deletion, or conservative substitution of amino acids, but retain at least one biological activity. Variant may also mean a protein with an amino acid sequence that is substantially identical to a referenced protein with an amino acid sequence that retains at least one biological activity. Representative examples of “biological activity” include the ability to be bound by a specific antibody or polypeptide or to promote an immune response. Variant can mean a functional fragment thereof. Variant can also mean multiple copies of a polypeptide. The multiple copies can be in tandem or separated by a linker. A conservative substitution of an amino acid, for example, replacing an amino acid with a different amino acid of similar properties (for example, hydrophilicity, degree and distribution of charged regions) is recognized in the art as typically involving a minor change. These minor changes may be identified, in part, by considering the hydropathic index of amino acids, as understood in the art (Kyte et al., J. Mol. Biol.1982, 157, 105-132, incorporated herein by reference). The hydropathic index of an amino acid is based on a consideration of its hydrophobicity and charge. It is known in the art that amino acids of similar hydropathic indexes may be substituted and still retain protein function. In one aspect, amino acids having hydropathic indexes of ±2 are substituted. The hydrophilicity of amino acids may also be used to reveal substitutions that would result in proteins retaining biological function. A consideration of the hydrophilicity of amino acids in the context of a peptide permits calculation of the greatest local average hydrophilicity of that peptide. Substitutions may be performed with amino acids having hydrophilicity values within ±2 of each other. Both the hydrophobicity index and the hydrophilicity value of amino acids are influenced by the particular side chain of that amino acid. Consistent with that observation, amino acid substitutions that are compatible with biological function are understood to depend on the relative similarity of the amino acids, and particularly the side chains of those amino acids, as revealed by the hydrophobicity, hydrophilicity, charge, size, and other properties. A variant can be an amino acid sequence that is substantially identical over the full length of the amino acid sequence or fragment thereof. The amino acid sequence can be 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%,Docket No.028193-0048-WO01 / 8355 95%, 96%, 97%, 98%, or 99%, or less than 100% identical over the full length of the amino acid sequence or a fragment thereof.

[0035] Substitutional variants typically contain the exchange of one amino acid for another at one or more sites within the protein, and may be designed to modulate one or more properties of the polypeptide, with or without the loss of other functions or properties. Substitutions may be conservative, that is, one amino acid is replaced with one of similar shape and charge. Conservative substitutions are well known in the art and include, for example, the changes of: alanine to serine; arginine to lysine; asparagine to glutamine or histidine; aspartate to glutamate; cysteine to serine; glutamine to asparagine; glutamate to aspartate; glycine to proline; histidine to asparagine or glutamine; isoleucine to leucine or valine; leucine to valine or isoleucine; lysine to arginine; methionine to leucine or isoleucine; phenylalanine to tyrosine, leucine or methionine; serine to threonine; threonine to serine; tryptophan to tyrosine; tyrosine to tryptophan or phenylalanine; and valine to isoleucine or leucine. Alternatively, substitutions may be non-conservative such that a function or activity of the polypeptide is affected. Non-conservative changes typically involve substituting a residue with one that is chemically dissimilar, such as a polar or charged amino acid for a nonpolar or uncharged amino acid, and vice versa.

[0036] Unless otherwise defined herein, scientific and technical terms used in connection with the present disclosure shall have the meanings that are commonly understood by those of ordinary skill in the art. For example, any nomenclatures used in connection with, and techniques of, cell and tissue culture, molecular biology, immunology, microbiology, genetics, and protein and nucleic acid chemistry and hybridization described herein are those that are well known and commonly used in the art. The meaning and scope of the terms should be clear; in the event however of any latent ambiguity, definitions provided herein take precedent over any dictionary or extrinsic definition. Further, unless otherwise required by context, singular terms shall include pluralities and plural terms shall include the singular. 2. Allergen Conjugate Peptide

[0037] Provided herein is an allergen conjugate peptide. The allergen conjugate peptide may include a self-assembling peptide and at least one allergen epitope attached thereto. As detailed herein, the allergen conjugate peptides including at least one allergen epitope, or a nanofiber comprising the same, may be used to treat an allergy or inflammation.Docket No.028193-0048-WO01 / 8355

[0038] Upon administration of the allergen conjugate peptide, or a nanofiber comprising the same, to a cell or subject, a variety of effects may be elicited in the cell or subject. In some embodiments, the allergen does not leak into the vascular compartment or bloodstream. The subject may form allergen-specific IgG antibodies. The allergen-specific IgG antibodies may be long-lasting. The allergen-specific IgG antibodies may be sustained. For example, the allergen-specific IgG antibodies may be detected in the subject for at least 20 weeks, at least 21 weeks, at least 22 weeks, at least 23 weeks, at least 24 weeks, at least 25 weeks, at least 26 weeks, at least 27 weeks, at least 28 weeks, at least 29 weeks, at least 30 weeks, at least 31 weeks, at least 32 weeks, at least 33 weeks, at least 34 weeks, at least 35 weeks, at least 36 weeks, at least 37 weeks, at least 38 weeks, at least 39 weeks, at least 40 weeks, at least 41 weeks, at least 42 weeks, at least 43 weeks, at least 44 weeks, at least 45 weeks, at least 46 weeks, at least 47 weeks, at least 48 weeks, at least 49 weeks, at least 50 weeks, at least 51 weeks, at least 52 weeks, at least 53 weeks, at least 54 weeks, or at least 55 weeks. The subject may become desensitized to the allergen. The subject may have a reduced Type I hypersensitivity reaction to the allergen. Activation of bone marrow-derived mast cells (BMMC) in the subject may be reduced. a. Self-Assembling Peptide

[0039] The compositions and methods detailed herein include self-assembling peptides. As used herein, the term “self-assembling peptide” refers to peptides that are able to spontaneously associate and form stable structures. Each self-assembling peptide may comprise or form an alpha helix. In other embodiments, each self-assembling peptide may comprise or form a beta-sheet. Examples of self-assembling peptides are detailed in, for example, U.S. Patent No.9,241,987; U.S. Patent No.9,849,174; U.S. Patent No. 10,596,238; U.S. Patent No.11,246,924; International Patent Application Publication No. WO 2023 / 044163; Lee, S. et al. Int. J. Mol. Sci.2019, 20, 5850; Hernandez, A. et al. Front. Bioeng. Biotechnol.2023, 11, 1139782; and Lopez-Silva et al. ACS Biomater. Sci. Eng. 2019, 5, 977-985, each of which is incorporated herein by reference in its entirety.

[0040] In some embodiments, the self-assembling peptide comprises a polypeptide having an amino acid sequence selected from bXXXb (SEQ ID NO: 1, wherein X is independently any amino acid and b is independently any positively charged amino acid), QQKFQFQFEQQ (SEQ ID NO: 12), FKFEFKFE (SEQ ID NO: 14), KFQFQFE (SEQ ID NO: 15), QQRFQFQFEQQ (SEQ ID NO: 16), QQRFQWQFEQQ (SEQ ID NO: 17), FEFEFKFKFEFEFKFK (SEQ ID NO: 18), QQRFEWEFEQQ (SEQ ID NO: 19), QQXFXWXFQQQ (SEQ ID NO: 20, where X is ornithine), FKFEFKFEFKFE (SEQ ID NO:Docket No.028193-0048-WO01 / 8355 21), FKFQFKFQFKFQ (SEQ ID NO: 22), AEAKAEAKAEAKAEAK (SEQ ID NO: 23), AEAEAKAKAEAEAKAK (SEQ ID NO: 24), AEAEAEAEAKAKAKAK (SEQ ID NO: 25), RADARADARADARADA (SEQ ID NO: 26), RARADADARARADADA (SEQ ID NO: 27), SGRGYBLGGQGAGAAAAAGGAGQGGYGGLGSQG (SEQ ID NO: 28), EWEXEXEXEX (SEQ ID NO: 29, where X is Val, Ala, Ser, or Pro), WKXKXKXKXK (SEQ ID NO: 30, where X is Val, Ala, Ser, or Pro), KWKVKVKVKVKVKVK (SEQ ID NO: 31, where X is Val, A, Ser, or Pro), LLLLKKKKKKKKLLLL (SEQ ID NO: 32), VKVKVKVKVDPPTKVKVKVKV (SEQ ID NO: 33), VKVKVKVKVDPPTKVKTKVKV (SEQ ID NO: 34), KVKVKVKVKDPPSVKVKVKVK (SEQ ID NO: 35), VKVKVKVKVDPPSKVKVKVKV (SEQ ID NO: 36), VKVKVKTKVDPPTKVKTKVKV (SEQ ID NO: 37), QQKFxFQFEQQ (SEQ ID NO: 38, wherein x is Glu, Asp, or Asn), QQKFQxQFEQQ (SEQ ID NO: 39, wherein x is Trp or Tyr), and QQKFQFxFEQQ (SEQ ID NO: 40, wherein x is Glu, Asp, or Asn), or a polypeptide with at least 75%, 80%, 85%, or 90% identity thereto. Some examples of self-assembling peptide are detailed in, for example, U.S. Patent Nos.9,241,987; 9,849,174; and 10,596,238, each of which is incorporated herein by reference in its entirety. In some embodiments, the self-assembling peptide comprises the amino acid sequence of QQKFQFQFEQQ (Q11, SEQ ID NO: 12), or a polypeptide with at least 75%, 80%, 85%, or 90% identity thereto. In some embodiments, the self-assembling peptide comprises thesequence Ac-QQKFQFQFEQQ-NH2(Q11, SEQ ID NO: 13), or a polypeptide with at least75%, 80%, 85%, or 90% identity thereto.

[0041] The self-assembling peptide may comprise an amino acid sequence of bXXXb (SEQ ID NO: 1), wherein X is independently any amino acid, and b is independently any positively charged amino acid. In such embodiments, each self-assembling peptide may form an alpha helix. In some embodiments, b is independently selected from Arg and Lys. In some embodiments, b is Arg. In some embodiments, b is Lys. In some embodiments, bXXXb (SEQ ID NO: 1) is RAYAR (SEQ ID NO: 2). In some embodiments, bXXXb (SEQ ID NO: 1) is KAYAK (SEQ ID NO: 3). In some embodiments, the self-assembling peptide comprises the sequence of RXXXR (SEQ ID NO: 4), wherein X is any amino acid. The self-assembling peptide may comprise an amino acid sequence of ZnbXXXbZm(SEQ ID NO: 5),wherein b is independently any positively charged amino acid, Z is independently any amino acid, X is independently any amino acid, n is an integer from 0 to 20, and m is an integer from 0 to 20. In some embodiments, n is an integer from 5 to 15, and m is an integer from 5 to 15. Some examples of self-assembling peptide are detailed in, for example, U.S. Patent No.11,246,924, which is incorporated herein by reference in its entirety. In such embodiments, a plurality of the conjugate peptides may assemble into a nanofiber.Docket No.028193-0048-WO01 / 8355

[0042] In some embodiments, the self-assembling peptide comprises a glutamine at the C-terminus. In some embodiments, the self-assembling peptide comprises a glutamine at the N-terminus. The self-assembling peptide may include at least, at most, or exactly 5, 10, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 40 amino acids. In some embodiments, the self-assembling peptide comprises 5 to 40 amino acids in length.

[0043] In some embodiments, the self-assembling peptide comprises an amino acid sequence of QARILEADAEILRAYARILEAHAEILRAQ (SEQ ID NO: 6) or QAKILEADAEILKAYAKILEAHAEILKAQ (SEQ ID NO: 7) or ADAEILRAYARILEAHAEILRAQ(SEQ ID NO: 8) or Ac-QARILEADAEILRAYARILEAHAEILRAQ-NH2(SEQ ID NO: 9) or Ac-QAKILEADAEILKAYAKILEAHAEILKAQ-NH2(SEQ ID NO: 10) or Ac-ADAEILRAYARILEAHAEILRAQ-NH2(SEQ ID NO: 11), or QAEILRAYARILEAQ (SEQ IDNO: 101), or QAEILRAYARILEAHAEILKAQ (SEQ ID NO: 102), or QAEILRAYARILEADAKILEAHAEILKAQ (SEQ ID NO: 103), or QAEILRAYARILEADAEILKAQAKILEAHAEILKAQ (SEQ ID NO: 104), or a polypeptide with at least 75%, 80%, 85%, 90%, or 95% identity thereto. In some embodiments, the self- assembling peptide comprises an amino acid sequence of QARILEADAEILRAYARILEAHAEILRAQ (SEQ ID NO: 6) or QAKILEADAEILKAYAKILEAHAEILKAQ (SEQ ID NO: 7) or ADAEILRAYARILEAHAEILRAQ (SEQ ID NO: 8) or a variant thereof. In some embodiments, the self-assembling peptide comprises an amino acid sequence of QARILEADAEILRAYARILEAHAEILRAQ (SEQ ID NO: 6). In some embodiments, the self-assembling peptide comprises an amino acid sequence of QAKILEADAEILKAYAKILEAHAEILKAQ (SEQ ID NO: 7). In some embodiments, the self- assembling peptide comprises an amino acid sequence of ADAEILRAYARILEAHAEILRAQ (SEQ ID NO: 8).

[0044] In some embodiments, each self-assembling peptide forms a beta sheet and comprises a polypeptide having an amino acid sequence selected from QQKFQFQFEQQ (SEQ ID NO: 12), FKFEFKFE (SEQ ID NO: 14), KFQFQFE (SEQ ID NO: 15), QQRFQFQFEQQ (SEQ ID NO: 16), QQRFQWQFEQQ (SEQ ID NO: 17), FEFEFKFKFEFEFKFK (SEQ ID NO: 18), QQRFEWEFEQQ (SEQ ID NO: 19), QQXFXWXFQQQ (SEQ ID NO: 20, where X is ornithine), FKFEFKFEFKFE (SEQ ID NO: 21), FKFQFKFQFKFQ (SEQ ID NO: 22), AEAKAEAKAEAKAEAK (SEQ ID NO: 23), AEAEAKAKAEAEAKAK (SEQ ID NO: 24), AEAEAEAEAKAKAKAK (SEQ ID NO: 25), RADARADARADARADA (SEQ ID NO: 26), RARADADARARADADA (SEQ ID NO: 27), SGRGYBLGGQGAGAAAAAGGAGQGGYGGLGSQG (SEQ ID NO: 28), EWEXEXEXEXDocket No.028193-0048-WO01 / 8355 (SEQ ID NO: 29, where X is Val, Ala, Ser, or Pro), WKXKXKXKXK (SEQ ID NO: 30, where X is Val, Ala, Ser, or Pro), KWKVKVKVKVKVKVK (SEQ ID NO: 31, where X is Val, A, Ser, or Pro), LLLLKKKKKKKKLLLL (SEQ ID NO: 32), VKVKVKVKVDPPTKVKVKVKV (SEQ ID NO: 33), VKVKVKVKVDPPTKVKTKVKV (SEQ ID NO: 34), KVKVKVKVKDPPSVKVKVKVK (SEQ ID NO: 35), VKVKVKVKVDPPSKVKVKVKV (SEQ ID NO: 36), VKVKVKTKVDPPTKVKTKVKV (SEQ ID NO: 37), QQKFxFQFEQQ (SEQ ID NO: 38, wherein x is Glu, Asp, or Asn), QQKFQxQFEQQ (SEQ ID NO: 39, wherein x is Trp or Tyr), and QQKFQFxFEQQ (SEQ ID NO: 40, wherein x is Glu, Asp, or Asn), or a polypeptide with at least 75%, 80%, 85%, or 90% identity thereto. In such embodiments, a plurality of the conjugate peptides may assemble into a nanofiber.

[0045] In some embodiments, the self-assembling peptide comprises a polypeptide having an amino acid sequence selected from VEVKVEVKV (SEQ ID NO: 41), VEVKVEVKVEVK (SEQ ID NO: 42), VVVAAAEEE (SEQ ID NO: 43), VEVEVEVEVEVEVEVEVEVE (SEQ ID NO: 44), CGNKRTRGC (SEQ ID NO: 45), VKVKVKVKVDPPTKVEVKVKV (SEQ ID NO: 46), LRKKLGKA (SEQ ID NO: 47), VVVVVVKK (SEQ ID NO: 48), AEAKAEAKAEAKAEAK (SEQ ID NO: 49), AEAKAEAK (SEQ ID NO: 50), AEAEAEAEAKAK (SEQ ID NO: 51), AEAEAKAK (SEQ ID NO: 52), AEAEAKAKAEAEAKAK (SEQ ID NO: 53), RADARADARADARADA (SEQ ID NO: 54), RADARGDARADARGDA (SEQ ID NO: 55), RADARADA (SEQ ID NO: 56), RARADADARARADADA (SEQ ID NO: 57), RARADADA (SEQ ID NO: 58), RARARARADADADADA (SEQ ID NO: 59), ADADADADARARARAR (SEQ ID NO: 60), DADADADARARARARA (SEQ ID NO: 61), RAEARAEARAEARAEA (SEQ ID NO: 62), RAEARAEA (SEQ ID NO: 63), KAKAKAKAEAEAEAEA (SEQ ID NO: 64), AEAEAEAEAKAKAKAK (SEQ ID NO: 65), KADAKADAKADAKADA (SEQ ID NO: 66), KADAKADA (SEQ ID NO: 67), AEAEAHAHAEAEAHAHA (SEQ ID NO: 68), AEAEAHAHA (SEQ ID NO: 69), HEHEHKHKHEHEHKHK (SEQ ID NO: 70), HEHEHKHK (SEQ ID NO: 71), FEFEFKFKFEFEFKFK (SEQ ID NO: 72), FEFKFEFK (SEQ ID NO: 73), LELELKLKLELELKLK (SEQ ID NO: 74), LELELKLK (SEQ ID NO: 75), KFDLKKDLKLDL (SEQ ID NO: 76), FKFEFKFF (SEQ ID NO: 77), FEFEFKFK (SEQ ID NO: 78), and RFRFRFRFRFRFRFRFRFRF (SEQ ID NO: 79), or a polypeptide with at least 75%, 80%, 85%, or 90% identity thereto. In such embodiments, a plurality of the PC-peptide conjugates may assemble into a nanofiber, nanotube, hydrogel, micelle, vesicle, nanoparticle, or suspension.

[0046] In some embodiments, the self-assembling polypeptide includes a modification to the C-terminus, to the N-terminus, or to both the C-terminus and N-terminus. N-terminalDocket No.028193-0048-WO01 / 8355 modifications may include, for example, biotin and acetyl. C-terminal modifications may include, for example, amide. In some embodiments, the N-terminus is acetylated (which may be indicated by “Ac” for example). In some embodiments, the C-terminus is amidated(which may be indicated by “NH2” for example, or “Am”).

[0047] The peptides described herein can be chemically synthesized using standard chemical synthesis techniques. In some embodiments the peptides are chemically synthesized by any of a number of fluid or solid phase peptide synthesis techniques known to those of skill in the art. Solid phase synthesis in which the C-terminal amino acid of the sequence is attached to an insoluble support followed by sequential addition of the remaining amino acids in the sequence is a preferred method for the chemical synthesis of the polypeptides described herein. Techniques for solid phase synthesis are well known to those of skill in the art and are described, for example, by Barany and Merrifield (1963) Solid-Phase Peptide Synthesis; pp.3-284 in The Peptides: Analysis, Synthesis, Biology. Vol. 2: Special Methods in Peptide Synthesis, Part A.; Merrifield et al. (1963) J. Am. Chem. Soc., 85: 2149-2156; and Stewart et al. (1984) Solid Phase Peptide Synthesis, 2nd ed. Pierce Chem. Co., Rockford, Ill; each of which is incorporated herein by reference. In some embodiments, the self-assembling peptide is synthesized by a solid phase peptide synthesis. b. Allergen Epitope

[0048] The allergen conjugate peptide includes at least one allergen epitope. The allergen epitope may be a B-cell epitope. The allergen epitope may be a T-cell epitope. The allergen epitope may be an epitope of a substance that a subject is allergic to. In some embodiments, the allergen epitope is IgE-reactive. In some embodiments, the allergen epitope comprises a peanut allergen epitope. The peanut allergen epitope may be from peanut allergen protein selected from Ara h 1, Ara h 3, or Ara h 7, or Ara h 2, or a combination thereof. The peanut allergen epitope may comprise the whole peanut allergen protein selected from Ara h 1, Ara h 3, or Ara h 7, or Ara h 2, or a portion thereof, or a combination thereof. The peanut allergen epitope may comprise AH1b (REREREEDWRQPREDWRRPS, SEQ ID NO: 106), AH3a (EDEYEYDEEDRRRGRGSRGR, SEQ ID NO: 107), AH7a (QEQDEYPYSRRGSRGRQPGE, SEQ ID NO: 108), AH1a (IDQIEKQAKDLAFPGSGE, SEQ ID NO: 109), AH2a (DPYSOSQDPYSOS, SEQ ID NO: 110, wherein O is hydroxyproline), or AH2b (DPYSOSDRRGAGSS, SEQ ID NO: 111, wherein O is hydroxyproline), or a combination thereof, or a peptide having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity thereto. TheDocket No.028193-0048-WO01 / 8355 peanut allergen epitope may be selected from AH1b (REREREEDWRQPREDWRRPS, SEQ ID NO: 106), AH3a (EDEYEYDEEDRRRGRGSRGR, SEQ ID NO: 107), AH7a (QEQDEYPYSRRGSRGRQPGE, SEQ ID NO: 108), AH1a (IDQIEKQAKDLAFPGSGE, SEQ ID NO: 109), AH2a (DPYSOSQDPYSOS, SEQ ID NO: 110, wherein O is hydroxyproline), or AH2b (DPYSOSDRRGAGSS, SEQ ID NO: 111, wherein O is hydroxyproline), or a combination thereof, or a peptide having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity thereto. In some embodiments, the peptide fibril is coupled to a plurality of allergen epitopes. The plurality of allergen epitopes coupled to a peptide fibril may be the same or different. Allergen epitope may be obtained commercially or synthesized and purified by any suitable method known in the art. In some embodiments, a peptide detailed herein includes C- terminal amidation. In some embodiments, a peptide detailed herein includes acetylation.

[0049] The allergen epitope may be conjugated or coupled to a self-assembling peptide by any means known in the art. The allergen epitope may be covalently coupled to a terminus of the self-assembling peptide. At least one allergen epitope may be attached to the C-terminus or the N-terminus of the self-assembling peptide. In some embodiments, the allergen epitope is covalently coupled to the N-terminus or N-terminal end of the self- assembling peptide. In some embodiments, the allergen epitope is covalently coupled to the C-terminus or C-terminal end of the self-assembling peptide. In some embodiments, the self-assembling peptide forms a beta sheet and the at least one allergen epitope is attached to the C-terminus or the N-terminus of the self-assembling peptide or a combination thereof. In some embodiments, the self-assembling peptide forms an alpha-helix and the at least one allergen epitope is attached to the N-terminus of the self-assembling peptide. The conjugation of the allergen epitope to the N-terminus or the N-terminal end of the self- assembling peptide may orient the allergen epitope towards the exterior of the helical peptide fibril once a plurality of allergen conjugate peptides assembles into a nanofiber. In some embodiments, the allergen epitopes are exposed on the exterior surface of the nanofiber. In some embodiments, the allergen epitopes are exposed on the exterior surface of the helical filament of the nanofiber.

[0050] The allergen conjugate peptide may include at least one allergen epitope. The allergen conjugate peptide may include 1 to 10 allergen epitopes attached to a self- assembling peptide. The allergen conjugate peptide may include 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 allergen epitopes attached to a self-assembling peptide. In embodiments including more than one allergen epitope attached to a self-assembling peptide, all allergen epitopes may be attached to the same end of the self-assembling peptide. For example, 1, 2, 3, or 4Docket No.028193-0048-WO01 / 8355 allergen epitopes together may be attached to the N-terminal end or to the C-terminal end of the self-assembling peptide. Once assembled into a nanofiber, the nanofiber may include n allergen epitopes, wherein n is an integer from 1 to 1,000, or 1 to 5,000, or 1 to 10,000, or 1 to 50,000, or 1 to 100,000, including all values and ranges there between. The allergen epitopes attached to a self-assembling peptide may be the same allergen epitope or different allergen epitopes.

[0051] In some embodiments, the antigens are exposed on the surface of the peptide fibril. In certain aspects, the ratio of antigen to self-assembling peptide is 1:1000, 1:100: 1:10, or 1:1, including all values and ranges there between. c. Linker

[0052] The allergen conjugate peptide may further comprise a linker. The linker may be between the allergen epitope and the self-assembling peptide. The linker may be between the at least one allergen epitope and the self-assembling peptide. In some embodiments, a linker is covalently attached to the self-assembling peptide between the allergen epitope and the self-assembling peptide. In some embodiments, the linker comprises at least one cysteine. The at least one cysteine may be at the N-terminus of the linker. In some embodiments, the linker comprises glycine and serine. In some embodiments, the linker comprises glycine and serine and cysteine. In some embodiments, the allergen epitope is attached to the self-assembling peptide through a thiol reactive group in the linker.

[0053] In some embodiments, the allergen conjugate peptide includes more than one linker. In such embodiments, the linkers may be the same or different from one another. The allergen conjugate peptide may include at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 linkers. The allergen conjugate peptide may include less than 20, less than 15, less than 10, or less than 5 linkers. The allergen conjugate peptide may include between 1 and 20, between 5 and 15, or between 1 and 5 linkers. The linker may be positioned at the C-terminus of the self- assembling peptide, at the N-terminus of the self-assembling peptide, or at both the N- and C-termini of the self-assembling peptide. In some embodiments, the linker is positioned at the N-terminus of the self-assembling peptide. Multiple linkers may be positioned adjacent to one another.

[0054] The linker may comprise, for example, an oligoethylene glycol, polyethylene glycol, or an amino acid sequence selected from SEQ ID NO: 83 (SGSG), SEQ ID NO: 84((Ser-Gly)2), SEQ ID NO: 85 (CCCCSGSG), SEQ ID NO: 86 (Gnwherein n is an integerDocket No.028193-0048-WO01 / 8355 from 1 to 10), SEQ ID NO: 87 (GSGS), SEQ ID NO: 88 (SSSS), SEQ ID NO: 89 (GGGS),SEQ ID NO: 90 (GGC), SEQ ID NO: 91 ((GGC)8), SEQ ID NO: 92 ((G4S)3), SEQ ID NO: 93(KSGSG), SEQ ID NO: 94 (KKSGSG), SEQ ID NO: 95 (EAAAK)2, SEQ ID NO: 96(GGAAY), and SGSG-C-SGSG (SEQ ID NO: 105). In some embodiments, the linkercomprises (Ser-Gly)2(SEQ ID NO: 85).d. PEG Molecule or PAS Peptide

[0055] The allergen conjugate peptide may comprise a polyethylene glycol (PEG) molecule. The self-assembling polypeptide may include a PEG molecule instead of an allergen epitope. A self-assembling polypeptide with a PEG molecule attached thereto may be referred to as a PEG conjugate peptide. Accordingly, provided herein is a PEG conjugate peptide, which may include a self-assembling polypeptide as detailed above with a PEG molecule attached thereto. The PEG molecule may facilitate mucus penetration. The PEG molecule may be muco-inert. The PEG molecule may enable or facilitate oral availability. The PEG molecule may be an average molecular weight from 1000 Da and up to and including 100,000 Da, or from 1000 Da and up to and including 5,000 Da. The PEG molecule may comprise PEG2000, PEG1000, or PEG3000. In some embodiments, thePEG molecule comprises PEG2000. The PEG molecule may comprise CH3O-(CH2CH2O)nwherein n is an integer from about 1 to about 2000, or from about 1 to about 100, or from about 20 to about 70. In some embodiments, the average n is at least about 20, about 45, or less than about 70. A plurality of conjugate peptides may have the same or different PEG molecule attached to each self-assembling peptide. The PEG molecule may be conjugated to a beta-sheet self-assembling peptide at the N-terminus or N-terminal end or at the C- terminus or C-terminal end or at both ends. The PEG molecule may be conjugated to a beta-sheet self-assembling peptide at the N-terminus or N-terminal end or at the C-terminus or C-terminal end, and include an allergen epitope conjugated at the other of the N-terminus or N-terminal end or the C-terminus or C-terminal end. The PEG molecule may be conjugated to the self-assembling peptide at the opposite terminus from wherein the allergen epitope is conjugated. The PEG molecule may be conjugated to an alpha-helical self- assembling peptide at the N-terminus or N-terminal end. The PEG molecule may be conjugated to the self-assembling peptide via a linker, as detailed above. The PEG molecule may be commercially obtained.

[0056] The allergen conjugate peptide may comprise a PAS peptide. The self- assembling polypeptide may include a PAS peptide instead of an allergen epitope. A self- assembling polypeptide with a PAS peptide attached thereto may be referred to as a PASDocket No.028193-0048-WO01 / 8355 conjugate peptide. Accordingly, provided herein is a PAS conjugate peptide, which may include a self-assembling polypeptide as detailed above with a PAS peptide attached thereto. The PAS peptide may be muco-inert. The PAS peptide may facilitate mucus penetration. The PAS peptide may enable or facilitate oral availability. The PAS peptide may comprise the amino acid sequence of Pro-Ala-Ser. The PAS peptide may comprise the amino acid sequence of ASPAAPAPASPAAPAPSAPA (SEQ ID NO: 97), or a peptide having at least 80%, 85%, 90%, or 95% identity thereto. In some embodiments, the PAS peptide comprises ASPAAPAPASPAAPAPSAPA (SEQ ID NO: 97). In some embodiments, the PASpeptide comprises H2N-ASPAAPAPASPAAPAPSAPA-NH2(SEQ ID NO: 98). The PASpeptide may be conjugated to a beta-sheet self-assembling peptide at the N-terminus or N- terminal end or at the C-terminus or C-terminal end or at both ends. The PAS peptide may be conjugated to a beta-sheet self-assembling peptide at the N-terminus or N-terminal end or at the C-terminus or C-terminal end, and include an allergen epitope conjugated at the other of the N-terminus or N-terminal end or the C-terminus or C-terminal end. The PAS peptide may be conjugated to the self-assembling peptide at the opposite terminus from wherein the allergen epitope is conjugated. The PAS peptide may be conjugated to an alpha-helical self-assembling peptide at the N-terminus or N-terminal end. The PAS peptide may be conjugated to the self-assembling peptide via a linker, as detailed above.

[0057] In some embodiments, the allergen conjugate peptide does not include a PEG molecule or a PAS peptide. In some such embodiments, the self-assembling peptide and / or the allergen epitope may have a hydrophobicity index such that the allergen conjugate peptide has sufficient activity or effectiveness without a PEG molecule or a PAS peptide. In some embodiments, a more hydrophobic epitope may need more of the allergen conjugate peptides in a nanofiber to be PASylated or PEGylated to have a desired activity or effectiveness. In some embodiments, a more hydrophillic epitope and / or self-assembling peptide may not need any or may need less of the allergen conjugate peptides in a nanofiber to be PASylated or PEGylated to have a desired activity or effectiveness.

[0058] The allergen conjugate peptide may comprise or consist of, for example, any one of SEQ ID NOs: 115-120 (as shown in TABLE 1), or a combination thereof. 3. T-cell Epitope Conjugate Peptides

[0059] Further provided herein is a T-cell epitope conjugate peptide. In some embodiments, a T-cell epitope may result in an IgG response against a target B-cell epitope. The T-cell epitope conjugate peptide may include a self-assembling peptide and at least one T-cell epitope attached thereto. The self-assembling peptide of the T-cell epitope conjugateDocket No.028193-0048-WO01 / 8355 peptide may be as detailed above for the allergen conjugate peptide. The T-cell epitope may be selected from PADRE and VAC. The PADRE molecule may comprise a polypeptide having the amino acid sequence of aKXVAAWTLKAa, wherein “X” comprises cyclohexylalanine and “a” comprises D-alanine (SEQ ID NO: 99). In some embodiments, the cyclohexylalanine comprises D-alanine. The VAC molecule may comprise a polypeptide having the amino acid sequence of QLVFNSISARALKAY (SEQ ID NO: 100).

[0060] In some embodiments, the T-cell epitope conjugate peptide further comprises a linker between the T-cell epitope and the self-assembling peptide. The linker of the T-cell epitope conjugate peptide may be as detailed above for the conjugate peptide. 4. Plain Self-Assembling Peptides

[0061] Some embodiments include plain self-assembling peptides, which refers to a self- assembling peptide as detailed above, but without any antigen such as an allergen epitope or a T-cell epitope (such as PADRE or VAC) or a PEG molecule or a PAS peptide attached thereto. 5. Nanofibers

[0062] Further described herein is a platform for vaccination or treatment based on peptides assembled into nanofibers. Provided herein is a nanofiber comprising a plurality of conjugate peptides, including allergen epitopes. The allergen conjugate peptides, with or without other conjugate peptides such as PEG conjugate peptides or PAS conjugate peptides or T-cell epitope conjugate peptides or plain self-assembling peptides, may self- assemble into a nanofiber. Self-assembly may refer to a process by which a plurality of conjugate peptides come together to form an organized structure such as a nanofiber or micelle or other larger form, based on interactions between conjugate peptides such as covalent, ionic, and Van der Waals interactions. For example, each type of conjugate peptide may be synthesized separately, and then multiple conjugate peptides may be mixed together in various ratios to form or assemble together into integrated nanofibers. The nanofiber may also be referred to as a peptide fibril. The nanofibers may be comprised of alpha-helical peptides. The nanofibers may be comprised of beta-sheet peptides. In this strategy, peptides fold into a complex beta-sheet-based or alpha-helix-based nanofiber where individual peptide coils run perpendicular to the axis of a long fibril. Each self- assembling peptide may comprise or form an alpha helix. The plurality of self-assembling peptides may form a peptide fibril in the form of a helical filament. The resultant nanostructure is composed of thousands of individual peptides or more. The self-Docket No.028193-0048-WO01 / 8355 assembling peptide may be extended N-terminally with a flexible spacer and an immune epitope such as an allergen epitope. In some embodiments, the nanofiber does not further comprise an adjuvant. In some embodiments, the nanofiber is an adjuvant.

[0063] Multiple epitope-bearing self-assembling peptides may be co-assembled into nanofibers composed of -sheets or -helices. Coiled coil folding requires more extensive design considerations compared to -sheet fibrillization, as both inter-helical interactions as well as those between the C-terminus and the main chain must be considered.

[0064] The nanofiber may be comprised of 10 to 10,000 peptides including all values and ranges there between. The nanofiber may include a combination of 10 to 10,000, or 100 to 10,000 conjugate peptides comprising allergen conjugate peptides. The nanofiber may comprise a combination of 10 to 10,000, or 100 to 10,000 peptides comprising allergen conjugate peptides, T-cell epitope conjugate peptides, and plain self-assembling peptides. The nanofiber may comprise a combination of 10 to 10,000, or 100 to 10,000 peptides comprising allergen conjugate peptides, PAS or PEG conjugate peptides, and plain self- assembling peptides. The nanofiber may comprise a combination of 10 to 10,000, or 100 to 10,000 peptides comprising allergen conjugate peptides, T-cell epitope conjugate peptides, PAS or PEG conjugate peptides, and plain self-assembling peptides.

[0065] The nanofiber may include at least one allergen conjugate peptide. The nanofiber may include a plurality of allergen conjugate peptides. The nanofiber may include at least 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250, 300, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1200, 1400, 1600, 1800, 2000, 2200, 2400, 2600, 2800, 3000, 3200, 3400, 3600, 3800, 4000, 4200, 4400, 4600, 4800, 5000, 5200, 5400, 5600, 5800, 6000, 6200, 6400, 6600, 6800, 7000, 7200, 7400, 7600, 7800, 8000, 8200, 8400, 8600, 8800, or 9000, or less than 10,000 of the same or different allergen conjugate peptides. The allergen conjugate peptides making up a single nanofiber may be the same or different. The nanofiber may further include at least one T-cell epitope conjugate peptide. The nanofiber may include a plurality of T-cell epitope conjugate peptides. The nanofiber may include at least 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250, 300, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1200, 1400, 1600, 1800, 2000, 2200, 2400, 2600, 2800, 3000, 3200, 3400, 3600, 3800, 4000, 4200, 4400, 4600, 4800, or 5000, or less than 5000 of the same or different T-cell epitope conjugate peptides. The T- cell epitope conjugate peptides making up a single nanofiber may be the same or different. The nanofiber may further include at least one T-cell epitope conjugate peptide. The nanofiber may include a plurality of PAS or PEG conjugate peptides. The nanofiber may include at least 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250, 300, 400, 450,Docket No.028193-0048-WO01 / 8355 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1200, 1400, 1600, 1800, 2000, 2200, 2400, 2600, 2800, 3000, 3200, 3400, 3600, 3800, 4000, 4200, 4400, 4600, 4800, or 5000, or less than 5000 of the same or different PAS or PEG conjugate peptides. The PAS or PEG conjugate peptides making up a single nanofiber may be the same or different. In some embodiments, the nanofiber includes self-assembling peptides without an allergen epitope conjugated thereto, which may be referred to as a plain self-assembling peptide. The nanofiber may further include at least one plain self-assembling peptide. The nanofiber may include a plurality of plain self-assembling peptides. The nanofiber may include at least 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250, 300, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1200, 1400, 1600, 1800, 2000, 2200, 2400, 2600, 2800, 3000, 3200, 3400, 3600, 3800, 4000, 4200, 4400, 4600, 4800, 5000, 5200, 5400, 5600, 5800, 6000, 6200, 6400, 6600, 6800, 7000, 7200, 7400, 7600, 7800, 8000, 8200, 8400, 8600, 8800, or 9000, or less than 10,000 of the same or different plain self-assembling peptides. The plain self-assembling peptides making up a single nanofiber may be the same or different.

[0066] A single nanofiber may include different peptides in a variety of ratios. The nanofiber may include a variety of ratios of different allergen epitopes. In some embodiments, at least about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 97.5% of the peptides in the nanofiber are allergen conjugate peptides. In some embodiments, at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, or 15% of the peptides in the nanofiber are T-cell epitope conjugate peptides. In some embodiments, at least about 1%, 10%, 12.5%, 15%, 17.5%, 20%, 22.5%, 25%, 27.5%, 30%, 32.5%, 35%, 37.5%, 40%, 42.5%, 45%, 47.5%, 50%, 52.5%, 55%, 57.5%, 60%, 62.5%, 65%, 67.5%, 70%, 72.5%, 75%, 77.5%, 80%, 82.5%, 85%, 87.5%, 90%, 92.5%, 95%, or 96.5%, or less than about 96.5% of the peptides in the nanofiber are PAS or PEG conjugate peptides. In some embodiments, at least about 1%, 10%, 12.5%, 15%, 17.5%, 20%, 22.5%, 25%, 27.5%, 30%, 32.5%, 35%, 37.5%, 40%, 42.5%, 45%, 47.5%, 50%, 52.5%, 55%, 57.5%, 60%, 62.5%, 65%, 67.5%, 70%, 72.5%, 75%, 77.5%, 80%, 82.5%, 85%, 87.5%, 90%, 92.5%, 95%, or 96.5%, or less than about 96.5% of the peptides in the nanofiber are plain self-assembling peptides. In some embodiments, the allergen peptide conjugate and the T-cell epitope peptide conjugate are present in the nanofiber at a ratio of about 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 12:1, 14:1, 16:1, 18:1, 20:1, 22:1, 24:1, 26:1, 28:1, 30:1, 32:1, 34:1, 36:1, 38:1, or 40:1.

[0067] The helical filament of the nanofiber may be formed around a central axis or core. The plurality of self-assembling peptides may form a peptide fibril in the form of a coiled coil.Docket No.028193-0048-WO01 / 8355 In some embodiments, the N-terminus of each self-assembling peptide is positioned at the exterior of the helical filament. The allergen epitopes may be exposed on the exterior surface of the nanofiber. An example of the self-assembling peptides formed into a peptide fibril is shown schematically in Egelman et al. (Structure 2015, 23, 280-289, incorporated herein by reference).

[0068] Nanofibers have been observed to be up to several microns long. The nanofiber can have a length of at least, at most, or exactly 100 nm, 200 nm, 300 nm, 400 nm, 500 nm, 600 nm, 700 nm, 800 nm, 900 nm, 1 μm, 1.2 μm, 1.4 μm, 1.6 μm, 1.8 μm, 2 μm, 2.2 μm, 2.4 μm, 2.6 μm, 2.8 μm, 3 μm, 3.2 μm, 3.4 μm, 3.6 μm, 3.8 μm, 4 μm, 4.2 μm, 4.4 μm, 4.6 μm, 4.8 μm, or 5 μm. The nanofiber may be about 100 nm to 1 μm, 100 nm to 2 μm, 100 nm to 3 μm, 100 nm to 4 μm, or 100 nm to 5 μm in length. The nanofiber can have a length of at least, at most, or exactly 0.01, 0.05, 0.1, 0.15, 0.20, 0.25, 0.5, 1, 5, 10, 20, 30, 40, 50, 75, 100, 125, 150, 175, 200, 225, 250, or 300 m, including all values and ranges there between. In some embodiments, the nanofiber is at least 100, 150, 200, 250, 300, or 350 nanometers in length. In some embodiments, the nanofiber is less than 10, 5, or 2 m in length. In some embodiments, the nanofiber is 50 nm to 600 nm in length. In certain aspects, the nanofiber has a molecular weight of at least 100, 500, 1,000, 5,000, 10,000,100,000 Da to 1 x 106, 1 x 107, 7 x 108Da, including all values and ranges there between.The nanofiber can have a diameter or width of at least, at most, or exactly 5, 10, 15, 20, 25, or 30 nm. In some embodiments, the nanofiber is 5-30 nm in diameter or width. a. Epitope Spacing in the Nanofiber

[0069] As detailed above, allergen-induced mast cell signaling through Fc RI-IgE isaffected by the distance that separates IgE-reactive epitopes, with more distant inter-epitope spacing yielding lower mast cell activation. B cell activation is enhanced by increasing antigen valency. However, in contrast to mast cells, B cell activation is maintained at distant inter-epitope spacings. Given this idiosyncratic sensitivity to epitope spacing between mast cells and B cells, the inventors discovered that immunogens bearing IgE-reactive B cell epitopes may be made hypoallergenic if the epitopes can be spaced far enough away from each other. The allergen epitopes in the nanofibers detailed herein may be spaced close together or far apart.

[0070] The spacing of the allergen epitopes in the nanofiber may be referred to as epitope density. The epitope density in the nanofiber may be high, low, or ultra low. For example, high epitope density may be a nanofiber being about 25% to about 100% epitope, or a nanofiber with about 25% to about 100% of the conjugate peptides having an epitopeDocket No.028193-0048-WO01 / 8355 attached thereto, or a nanofiber with about 25% to about 100% of the conjugate peptides being allergen conjugate peptides. High epitope density may be a nanofiber being about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 100% epitope, or a nanofiber with about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 100% of the conjugate peptides having an epitope attached thereto, or a nanofiber with about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 100% of the conjugate peptides being allergen conjugate peptides. Low epitope density may be a nanofiber being more than about 1% to less than about 25% epitope, or a nanofiber with more than about 1% to less than about 25% of the conjugate peptides having an epitope attached thereto, or a nanofiber with more than about 1% to less than about 25% of the conjugate peptides being allergen conjugate peptides. Low epitope density may be a nanofiber being more than about 1%, or about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 21%, about 22%, about 23%, about 24%, or less than about 25% epitope, or a nanofiber with more than about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 21%, about 22%, about 23%, about 24%, or less than about 25% of the conjugate peptides having an epitope attached thereto, or a nanofiber with more than about 1%, or about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 21%, about 22%, about 23%, about 24%, or less than about 25% of the conjugate peptides being allergen conjugate peptides. Ultra low epitope density may be a nanofiber being more than about 0% to about 1% epitope, or a nanofiber with more than about 0% to about 1% of the conjugate peptides having an epitope attached thereto, or a nanofiber with more than about 0% to about 1% of the conjugate peptides being allergen conjugate peptides. Ultra low epitope density may be a nanofiber being more than about 0%, or more than about 0.05%, or about 0.05%, about 0.06%, about 0.07%, about 0.08%, about 0.09%, about 0.1%, about 0.2%, about 0.3%, about 0.4%, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, or about 1% epitope, or a nanofiber with more than about 0%, or more than about 0.05%, or about 0.05%, about 0.06%, about 0.07%, about 0.08%, about 0.09%, about 0.1%, about 0.2%, about 0.3%, about 0.4%, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, orDocket No.028193-0048-WO01 / 8355 about 1% of the conjugate peptides having an epitope attached thereto, or a nanofiber with more than about 0%, or more than about 0.05%, or about 0.05%, about 0.06%, about 0.07%, about 0.08%, about 0.09%, about 0.1%, about 0.2%, about 0.3%, about 0.4%, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, or about 1% of the conjugate peptides being allergen conjugate peptides.

[0071] The spacing of the allergen epitopes in the nanofiber may be referred to as epitope spacing. The epitope spacing in the nanofiber may be high, low, or ultra low. Ultra low epitope spacing (high epitope density) may be about 0.22 nm to about 0.88 nm between adjacent epitopes. Ultra low epitope spacing may be about 0.22 nm, about 0.24 nm, about 0.26 nm, about 0.28 nm, about 0.30 nm, about 0.32 nm, about 0.34 nm, about 0.36 nm, about 0.38 nm, about 0.40 nm, about 0.42 nm, about 0.44 nm, about 0.46 nm, about 0.48 nm, about 0.50 nm, about 0.52 nm, about 0.54 nm, about 0.56 nm, about 0.58 nm, about 0.60 nm, about 0.62 nm, about 0.64 nm, about 0.66 nm, about 0.68 nm, about 0.70 nm, about 0.72 nm, about 0.74 nm, about 0.76 nm, about 0.78 nm, about 0.80 nm, about 0.82 nm, about 0.84 nm, about 0.86 nm, or about 0.88 nm between adjacent epitopes. Ultra low epitope spacing may be less than about 0.24 nm, less than about 0.26 nm, less than about 0.28 nm, less than about 0.30 nm, less than about 0.32 nm, less than about 0.34 nm, less than about 0.36 nm, less than about 0.38 nm, less than about 0.40 nm, less than about 0.42 nm, less than about 0.44 nm, less than about 0.46 nm, less than about 0.48 nm, less than about 0.50 nm, less than about 0.52 nm, less than about 0.54 nm, less than about 0.56 nm, less than about 0.58 nm, less than about 0.60 nm, less than about 0.62 nm, less than about 0.64 nm, less than about 0.66 nm, less than about 0.68 nm, less than about 0.70 nm, less than about 0.72 nm, less than about 0.74 nm, less than about 0.76 nm, less than about 0.78 nm, less than about 0.80 nm, less than about 0.82 nm, less than about 0.84 nm, less than about 0.86 nm, or less than about 0.88 nm between adjacent epitopes. Ultra low epitope spacing may be more than about 0.22 nm, more than about 0.24 nm, more than about 0.26 nm, more than about 0.28 nm, more than about 0.30 nm, more than about 0.32 nm, more than about 0.34 nm, more than about 0.36 nm, more than about 0.38 nm, more than about 0.40 nm, more than about 0.42 nm, more than about 0.44 nm, more than about 0.46 nm, more than about 0.48 nm, more than about 0.50 nm, more than about 0.52 nm, more than about 0.54 nm, more than about 0.56 nm, more than about 0.58 nm, more than about 0.60 nm, more than about 0.62 nm, more than about 0.64 nm, more than about 0.66 nm, more than about 0.68 nm, more than about 0.70 nm, more than about 0.72 nm, more than about 0.74 nm, more than about 0.76 nm, more than about 0.78 nm, more than about 0.80 nm, more than about 0.82 nm, more than about 0.84 nm, or more than about 0.86 nm between adjacent epitopes.Docket No.028193-0048-WO01 / 8355

[0072] Low epitope spacing (low epitope density) may be more than about 0.88 nm to less than about 22 nm between adjacent epitopes. Low epitope spacing may be more than about 0.88 nm, or about 0.9 nm, about 1 nm, about 2 nm, about 3 nm, about 4 nm, about 5 nm, about 6 nm, about 7 nm, about 8 nm, about 9 nm, about 10 nm, about 11 nm, about 12 nm, about 13 nm, about 14 nm, about 15 nm, about 16 nm, about 17 nm, about 18 nm, about 19 nm, about 20 nm, about 21 nm, or less than about 22 nm between adjacent epitopes. Low epitope spacing may be less than about 0.9 nm, less than about 1 nm, less than about 2 nm, less than about 3 nm, less than about 4 nm, less than about 5 nm, less than about 6 nm, less than about 7 nm, less than about 8 nm, less than about 9 nm, less than about 10 nm, less than about 11 nm, less than about 12 nm, less than about 13 nm, less than about 14 nm, less than about 15 nm, less than about 16 nm, less than about 17 nm, less than about 18 nm, less than about 19 nm, less than about 20 nm, less than about 21 nm, or less than about 22 nm between adjacent epitopes. Low epitope spacing may be more than about 0.88 nm, more than about 0.9 nm, more than about 1 nm, more than about 2 nm, more than about 3 nm, more than about 4 nm, more than about 5 nm, more than about 6 nm, more than about 7 nm, more than about 8 nm, more than about 9 nm, more than about 10 nm, more than about 11 nm, more than about 12 nm, more than about 13 nm, more than about 14 nm, more than about 15 nm, more than about 16 nm, more than about 17 nm, more than about 18 nm, more than about 19 nm, more than about 20 nm, or more than about 21 nm between adjacent epitopes.

[0073] High epitope spacing (ultra low epitope density) may be more than about 22 nm between adjacent epitopes. High epitope spacing may be more than about 22 nm to about 450 nm between adjacent epitopes. High epitope spacing may be more than about 22 nm, about 25 nm, about 30 nm, about 35 nm, about 40 nm, about 45 nm, about 50 nm, about 55 nm, about 60 nm, about 65 nm, about 70 nm, about 75 nm, about 80 nm, about 85 nm, about 90 nm, about 95 nm, about 100 nm, about 150 nm, about 200 nm, about 250 nm, about 300 nm, about 350 nm, about 400 nm, or about 450 nm between adjacent epitopes. High epitope spacing may be more than about 22 nm, or less than about 25 nm, less than about 30 nm, less than about 35 nm, less than about 40 nm, less than about 45 nm, less than about 50 nm, less than about 55 nm, less than about 60 nm, less than about 65 nm, less than about 70 nm, less than about 75 nm, less than about 80 nm, less than about 85 nm, less than about 90 nm, less than about 95 nm, less than about 100 nm, less than about 150 nm, less than about 200 nm, less than about 250 nm, less than about 300 nm, less than about 350 nm, less than about 400 nm, or less than about 450 nm between adjacent epitopes. High epitope spacing may be more than about 22 nm, more than about 25 nm, more than about 30 nm, more than about 35 nm, more than about 40 nm, more than aboutDocket No.028193-0048-WO01 / 8355 45 nm, more than about 50 nm, more than about 55 nm, more than about 60 nm, more than about 65 nm, more than about 70 nm, more than about 75 nm, more than about 80 nm, more than about 85 nm, more than about 90 nm, more than about 95 nm, more than about 100 nm, more than about 150 nm, more than about 200 nm, more than about 250 nm, more than about 300 nm, more than about 350 nm, more than about 400 nm, or more than about 450 nm between adjacent epitopes.

[0074] For example, adjacent allergen epitopes in the nanofiber may have more than about 0.22 nm, more than about 0.88 nm, about 0.9 nm to about 22 nm, about 10 nm to about 20 nm, more than about 22 nm, less than about 440 nm, more than about 0.22 nm to less than about 440 nm, more than about 0.88 nm to less than about 440 nm, more than about 22 nm to less than about 440 nm, about 25 nm to about 30 nm, or about 28 nm between them. Adjacent allergen epitopes in the nanofiber may have about 25 nm to about 30 nm, about 26 nm to 29 nm, or about 28 nm between them. The inter-epitope spacing may be about 25 nm to about 30 nm, about 26 nm to about 29 nm, or about 28 nm.

[0075] Epitope spacing in the nanofiber may be achieved by mixing allergen conjugate peptides with self-assembling peptides without an allergen epitope attached thereto. Epitope spacing in the nanofiber may be achieved by mixing allergen conjugate peptides with plain self-assembling peptides. Epitope spacing in the nanofiber may be achieved by mixing allergen conjugate peptides with T-cell epitope conjugate peptides. Epitope spacing in the nanofiber may be achieved by mixing allergen conjugate peptides with PAS or PEG conjugate peptides. Epitope spacing in the nanofiber may be achieved by mixing allergen conjugate peptides with plain self-assembling peptides, or with T-cell epitope conjugate peptides, or with PAS or PEG conjugate peptides, or with a combination thereof. For example, about 0.1%, about 0.2%, about 0.3%, about 0.4%, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1%, about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 100% of the peptides in the nanofiber may be allergen conjugate peptides. At least about 0.1%, at least about 0.2%, at least about 0.3%, at least about 0.4%, at least about 0.5%, at least about 0.6%, at least about 0.7%, at least about 0.8%, at least about 0.9%, at least about 1%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, or at least about 95% of the peptides in the nanofiber may be allergenDocket No.028193-0048-WO01 / 8355 conjugate peptides. Less than about 0.1%, less than about 0.2%, less than about 0.3%, less than about 0.4%, less than about 0.5%, less than about 0.6%, less than about 0.7%, less than about 0.8%, less than about 0.9%, less than about 1%, less than about 5%, less than about 10%, less than about 15%, less than about 20%, less than about 25%, less than about 30%, less than about 35%, less than about 40%, less than about 45%, less than about 50%, less than about 55%, less than about 60%, less than about 65%, less than about 70%, less than about 75%, less than about 80%, less than about 85%, less than about 90%, less than about 95%, or less than about 100% of the peptides in the nanofiber may be allergen conjugate peptides. More than 0%, more than about 0.1%, more than about 0.2%, more than about 0.3%, more than about 0.4%, more than about 0.5%, more than about 0.6%, more than about 0.7%, more than about 0.8%, more than about 0.9%, more than about 1%, more than about 5%, more than about 10%, more than about 15%, more than about 20%, more than about 25%, more than about 30%, more than about 35%, more than about 40%, more than about 45%, more than about 50%, more than about 55%, more than about 60%, more than about 65%, more than about 70%, more than about 75%, more than about 80%, more than about 85%, more than about 90%, or more than about 95% of the peptides in the nanofiber may be allergen conjugate peptides. In some embodiments, at least about 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, or 1% of the peptides in the nanofiber may be conjugate peptides. The nanofiber may comprise, for example, about 0.2-1% conjugate peptides and 99-99.8% plain self-assembling peptides, or about 0.5% conjugate peptides and 99.5% plain self-assembling peptides. For example, more than about 0.05%, less than about 1%, less than about 25%, less than about 20%, at least about 0.05%, at least about 0.1%, at least about 0.2%, at least about 0.3%, at least about 0.4%, at least about 0.5%, at least about 0.6%, at least about 0.7%, at least about 0.8%, at least about 0.9%, at least about 1%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% of the peptides in the nanofiber are allergen conjugate peptides. 6. Immune Response and Immunoassays

[0076] As discussed above, the compositions and methods provided herein include evoking or inducing an immune response in a subject against an antigen. The antigen may comprise an allergen epitope. In one embodiment, the immune response can protect against or treat a subject having, suspected of having, or at risk of developing a disease.Docket No.028193-0048-WO01 / 8355 One use of the immunogenic compositions is to provide effective vaccines and / or active immunotherapies. The compositions detailed herein may induce an immune response. The immune response may be an antigen-specific immune response. In some embodiments, the antigen-specific immune response is temporary or not life-long. In some embodiments, the antigen-specific immune response is long lasting. “Long lasting” may be an antibody response that lasts for at least about 2 weeks, at least about 3 weeks, at least about 4 weeks, at least about 6 weeks, at least about 10 weeks, at least about 12 weeks, at least about 14 weeks, at least about 16 weeks, at least about 18 weeks, at least about 20 weeks, at least about 22 weeks, at least about 24 weeks, at least about 26 weeks, at least about 28 weeks, at least about 30 weeks, at least about 32 weeks, at least about 34 weeks, at least about 36 weeks, at least about 38 weeks, at least about 40 weeks, at least about 42 weeks, at least about 44 weeks, at least about 46 weeks, at least about 48 weeks, at least about 50 weeks, at least about 52 weeks, at least about 1 year, at least about 1.5 years, at least about 2 years, at least about 2.5 years, at least about 3 years, at least about 3.5 years, at least about 4 years, at least about 4.5 years, at least about 5 years, at least about 5.5 years, at least about 6 years, at least about 7 years, at least about 8 years, at least about 9 years, at least about 10 years, at least about 15 years, at least about 20 years, at least about 25 years, at least about 30 years, at least about 35 years, at least about 40 years, at least about 45 years, at least about 50 years, at least about 55 years, at least about 60 years, at least about 65 years, at least about 70 years, at least about 75 years, at least about 80 years, at least about 85 years, at least about 90 years, or at least about 95 years, or a lifetime. “Long lasting” may be an antibody response that lasts for about 3 weeks to a lifetime, about 1 month to a lifetime, about 1 year to a lifetime, about 1 year to about 30 years, or about 5 years to about 20 years. The anti-allergen antibodies generated may be IgM, or IgG, or IgA, or a combination thereof. In some embodiments, the antibodies are IgM antibodies. In some embodiments, the antibodies are IgA antibodies. In some embodiments, the antibodies are IgG antibodies. The immune response may include IgG antibody isotypes and subclasses. In some embodiments, the immune response comprises IgG1, IgG2, IgG3, or IgG4 antibody isotypes, or a combination thereof. In some embodiments, the immune response comprises IgM antibody isotypes. In some embodiments, the immune response comprises IgG antibody isotypes. The immunogenic composition may have increased immunogenicity relative to a control. In some embodiments, the control comprises an allergen epitope or other antigen without a self-assembling peptide.

[0077] The compositions and methods detailed herein may increase the level of anti- allergen antibodies relative to a control. The level of anti-allergen antibodies may be increased by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%,Docket No.028193-0048-WO01 / 8355 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7- fold, 8-fold, 9-fold, or 10-fold, relative to a control. The level of anti-allergen antibodies may be increased by less than about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7- fold, 8-fold, 9-fold, or 10-fold, relative to a control. The level of anti-allergen antibodies may be increased by about 5-95%, 10-90%, 15-85%, 20-80%, or 1.5-fold to 10-fold, relative to a control. In some embodiments, IgG antibodies may be increased. The compositions and methods detailed herein may decrease the level of anti-allergen antibodies relative to a control. The level of anti-allergen antibodies may be decreased by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, or 10-fold, relative to a control. The level of anti-allergen antibodies may be decreased by less than about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, or 10-fold, relative to a control. The level of anti-allergen antibodies may be decreased by about 5-95%, 10-90%, 15-85%, 20-80%, or 1.5-fold to 10-fold, relative to a control. In some embodiments, IgE antibodies may be decreased.

[0078] Further provided herein is the implementation of serological assays to evaluate whether and to what extent an immune response is induced or evoked by compositions. There are many types of immunoassays that can be implemented. Immunoassays include, but are not limited to, those described in U.S. Patent No.4,367,110 (double monoclonal antibody sandwich assay) and U.S. Patent No.4,452,901 (western blot), which are incorporated herein by reference. Other assays include immunoprecipitation of labeled ligands and immunocytochemistry, both in vitro and in vivo.

[0079] Immunoassays generally are binding assays. Certain immunoassays are the various types of enzyme linked immunosorbent assays (ELISAs) and radioimmunoassays (RIA) known in the art. Immunohistochemical detection using tissue sections may also be useful. In one example, antibodies or antigens are immobilized on a selected surface, such as a well in a polystyrene microtiter plate, dipstick, or column support. Then, a test composition suspected of containing the desired antigen or antibody, such as a clinical sample, is added to the wells. After binding and washing to remove non-specifically bound immune complexes, the bound antigen or antibody may be detected. Detection is generally achieved by the addition of another antibody, specific for the desired antigen or antibody, that is linked to a detectable label. This type of ELISA is known as a “sandwich ELISA.” Detection also may be achieved by the addition of a second antibody specific for the desiredDocket No.028193-0048-WO01 / 8355 antigen, followed by the addition of a third antibody that has binding affinity for the second antibody, with the third antibody being linked to a detectable label.

[0080] Competition ELISAs are also possible implementations in which test samples compete for binding with known amounts of labeled antigens or antibodies. The amount of reactive species in the unknown sample is determined by mixing the sample with the known labeled species before or during incubation with coated wells. The presence of reactive species in the sample acts to reduce the amount of labeled species available for binding to the well and thus reduces the ultimate signal. Irrespective of the format employed, ELISAs have certain features in common, such as coating, incubating or binding, washing to remove non-specifically bound species, and detecting the bound immune complexes.

[0081] Antigen or antibodies may also be linked to a solid support, such as in the form of plate, beads, dipstick, membrane, or column matrix, and the sample to be analyzed is applied to the immobilized antigen or antibody. In coating a plate with either antigen or antibody, one will generally incubate the wells of the plate with a solution of the antigen or antibody, either overnight or for a specified period. The wells of the plate will then be washed to remove incompletely-adsorbed material. Any remaining available surfaces of the wells are then “coated” with a nonspecific protein that is antigenically neutral with regard to the test antisera. These include bovine serum albumin (BSA), casein, and solutions of milk powder. The coating allows for blocking of nonspecific adsorption sites on the immobilizing surface and thus reduces the background caused by nonspecific binding of antisera onto the surface. a. Protective Immunity

[0082] In some embodiments, proteinaceous compositions confer protective immunity to a subject. Protective immunity refers to a body’s ability to mount a specific immune response that protects the subject from developing a particular disease or condition that involves the agent against which there is an immune response. An immunogenically effective amount is capable of conferring protective immunity to the subject.

[0083] As used herein the phrase “immune response” or its equivalent “immunological response” may refer to the development of a humoral (antibody mediated), cellular (mediated by antigen-specific T cells or their secretion products) or both humoral and cellular response directed against a protein, peptide, carbohydrate, or polypeptide in a recipient patient. Such a response can be an active response induced by administration of immunogen or a passive response induced by administration of antibody, antibodyDocket No.028193-0048-WO01 / 8355 containing material, or primed T-cells. A cellular immune response is elicited by the presentation of polypeptide epitopes in association with Class I or Class II MHC molecules, to activate antigen-specific CD4 (+) T helper cells and / or CD8 (+) cytotoxic T cells. The response may also involve activation of monocytes, macrophages, NK cells, basophils, dendritic cells, astrocytes, microglia cells, eosinophils or other components of innate immunity. As used herein “active immunity” refers to any immunity conferred upon a subject by administration of an antigen.

[0084] As used herein “passive immunity” refers to any immunity conferred upon a subject without administration of an antigen to the subject. “Passive immunity” therefore includes, but is not limited to, administration of activated immune effectors including cellular mediators or protein mediators (e.g., monoclonal and / or polyclonal antibodies) of an immune response. A monoclonal or polyclonal antibody composition may be used in passive immunization for the prevention or treatment of infection by organisms that carry the antigen recognized by the antibody. An antibody composition may include antibodies that bind to a variety of antigens that may in turn be associated with various organisms. The antibody component can be a polyclonal antiserum. In certain aspects the antibody or antibodies are affinity purified from an animal or second subject that has been challenged with an antigen(s). Alternatively, an antibody mixture may be used, which is a mixture of monoclonal and / or polyclonal antibodies to antigens for the same, related, or different disease or disorder.

[0085] Passive immunity may be imparted to a patient or subject by administering to the patient immunoglobulins (Ig) and / or other immune factors obtained from a donor or other non-patient source having a known immunoreactivity. In other aspects, an antigenic composition as detailed herein can be administered to a subject who then acts as a source or donor for globulin, produced in response to challenge with the antigenic composition (“hyperimmune globulin”), that contains antibodies directed against an allergen epitope, for example. A subject thus treated would donate plasma from which hyperimmune globulin would then be obtained, via conventional plasma-fractionation methodology, and administered to another subject in order to impart resistance against or to treat inflammation, for example. Hyperimmune globulins are particularly useful for immune-compromised individuals, for individuals undergoing invasive procedures or where time does not permit the individual to produce their own antibodies in response to vaccination. See U.S. Patent Nos. 6,936,258, 6,770,278, 6,756,361, 5,548,066, 5,512,282, 4,338,298, and 4,748,018, each of which is incorporated herein by reference in its entirety, for exemplary methods and compositions related to passive immunity.Docket No.028193-0048-WO01 / 8355

[0086] For purposes of this specification and the accompanying claims the terms “epitope” and “antigenic determinant” are used interchangeably to refer to a site on an antigen to which B and / or T cells respond or recognize. B-cell epitopes can be formed from small molecules. For example, B-cell epitopes may be formed from an allergen epitope. B- cell epitopes can be formed both from contiguous amino acids or noncontiguous amino acids juxtaposed by tertiary folding of a protein. Epitopes formed from contiguous amino acids are typically retained on exposure to denaturing solvents whereas epitopes formed by tertiary folding are typically lost on treatment with denaturing solvents. An epitope typically includes at least 3, and more usually, at least 5 or 8-10 amino acids in a unique spatial conformation. Methods of determining spatial conformation of epitopes include, for example, x-ray crystallography and 2-dimensional nuclear magnetic resonance. See, e.g., Epitope Mapping Protocols (1996), incorporated herein by reference. Antibodies that recognize the same epitope can be identified in a simple immunoassay showing the ability of one antibody to block the binding of another antibody to a target antigen. T-cells recognize continuous epitopes of about nine amino acids for CD8 cells or about 13-15 amino acids for CD4 cells. T cells that recognize the epitope can be identified by in vitro assays that measure antigen-dependent proliferation, as determined by3H-thymidine incorporation by primed T cells inresponse to an epitope (Burke et al., 1994, incorporated herein by reference), by antigen- dependent killing (cytotoxic T lymphocyte assay, Tigges et al., 1996, incorporated herein by reference) or by cytokine secretion.

[0087] The presence of a cell-mediated immunological response can be determined by proliferation assays (CD4 (+) T cells) or CTL (cytotoxic T lymphocyte) assays. The relative contributions of humoral and cellular responses to the protective or therapeutic effect of an immunogen can be distinguished by separately isolating IgG and T-cells from an immunized syngeneic animal and measuring protective or therapeutic effect in a second subject.

[0088] As used herein and in the claims, the terms “antibody” or “immunoglobulin” are used interchangeably and refer to any of several classes of structurally related proteins that function as part of the immune response of an animal or recipient, which proteins include IgG, IgD, IgE, IgA, IgM, and related proteins.

[0089] Under normal physiological conditions antibodies are found in plasma and other body fluids and in the membrane of certain cells and are produced by lymphocytes of the type denoted B cells or their functional equivalent. Antibodies of the IgG class are made up of four polypeptide chains linked together by disulfide bonds. The four chains of intact IgGDocket No.028193-0048-WO01 / 8355 molecules are two identical heavy chains referred to as H-chains and two identical light chains referred to as L-chains.

[0090] In order to produce polyclonal antibodies, a host, such as a rabbit or goat, is immunized with the antigen or antigen fragment, generally with an adjuvant and, if necessary, coupled to a carrier. Antibodies to the antigen are subsequently collected from the sera of the host. The polyclonal antibody can be affinity purified against the antigen rendering it monospecific. Monoclonal antibodies can be produced by hyperimmunization of an appropriate donor with the antigen or ex-vivo by use of primary cultures of splenic cells or cell lines derived from spleen (Anavi, 1998; Huston et al., 1991; Johnson et al., 1991; Mernaugh et al., 1995, each incorporated herein by reference).

[0091] As used herein, the phrase “an immunological portion of an antibody” includes a Fab fragment of an antibody, a Fv fragment of an antibody, a heavy chain of an antibody, a light chain of an antibody, a heterodimer consisting of a heavy chain and a light chain of an antibody, a variable fragment of a light chain of an antibody, a variable fragment of a heavy chain of an antibody, and a single chain variant of an antibody, which is also known as scFv. In addition, the term includes chimeric immunoglobulins which are the expression products of fused genes derived from different species, one of the species can be a human, in which case a chimeric immunoglobulin is said to be humanized. Typically, an immunological portion of an antibody competes with the intact antibody from which it was derived for specific binding to an antigen.

[0092] Optionally, an antibody or preferably an immunological portion of an antibody, can be chemically conjugated to, or expressed as, a fusion protein with other proteins. For purposes of this specification and the accompanying claims, all such fused proteins are included in the definition of antibodies or an immunological portion of an antibody. 7. Pharmaceutical Compositions

[0093] Further provided herein are pharmaceutical compositions comprising the allergen peptide conjugate or nanofibers detailed herein. Compositions can include a peptide fibril coupled to a plurality of antigens such as an allergen epitope, and may be referred to as a “fibril complex.” In some embodiments, the composition does not further comprise an adjuvant. In some embodiments, the composition further comprises an adjuvant. In some embodiments, the peptide fibril is an adjuvant.

[0094] The preparation of pharmaceutical compositions such as vaccines that contain polypeptide or peptide sequence(s) as active ingredients is generally well understood in theDocket No.028193-0048-WO01 / 8355 art, as exemplified by U.S. Patent Nos.4,608,251; 4,601,903; 4,599,231; 4,599,230; 4,596,792; and 4,578,770, all of which are incorporated herein by reference. Typically, such pharmaceutical compositions are prepared as injectables either as liquid solutions or suspensions: solid forms suitable for solution in or suspension in liquid prior to injection may also be prepared. The preparation may also be emulsified. The active immunogenic ingredient is often mixed with excipients that are pharmaceutically acceptable and compatible with the active ingredient. Suitable excipients are, for example, water, saline, dextrose, glycerol, ethanol, or the like and combinations thereof. In addition, if desired, the vaccine may contain amounts of auxiliary substances such as wetting or emulsifying agents, pH buffering agents, or adjuvants that enhance the effectiveness of the vaccines. In specific embodiments, pharmaceutical compositions are formulated with a combination of substances, as described in U.S. Patent Nos.6,793,923 and 6,733,754, which are incorporated herein by reference.

[0095] Pharmaceutical compositions may be conventionally administered parenterally, by injection, for example, either subcutaneously or intramuscularly. Additional formulations which are suitable for other modes of administration include suppositories and, in some cases, oral formulations. For suppositories, traditional binders and carriers may include, for example, polyalkalene glycols or triglycerides: such suppositories may be formed from mixtures containing the active ingredient in the range of about 0.5% to about 10%, preferably about 1% to about 2%. Oral formulations include such normally employed excipients as, for example, pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose, magnesium carbonate and the like. These compositions take the form of solutions, suspensions, tablets, pills, capsules, sustained release formulations or powders and contain about 10% to about 95% of active ingredient, preferably about 25% to about 70%.

[0096] In some embodiments, the compositions described herein are formulated for sublingual administration. The compositions may be formulated into a tablet. For example, the compositions may be formulation into a dissolving tablet. The dissolving tablet may be placed under the tongue. The compositions may be formulated into a table by a variety of suitable means, for example, the compositions may be freeze dried and pressed into a tablet form.

[0097] The compositions described herein may be formulated into a pharmaceutical composition as neutral or salt forms. Pharmaceutically-acceptable salts include the acid addition salts (formed with the free amino groups of the peptide) and those that are formed with inorganic acids such as, for example, hydrochloric or phosphoric acids, or such organicDocket No.028193-0048-WO01 / 8355 acids as acetic, oxalic, tartaric, mandelic, and the like. Salts formed with the free carboxyl groups may also be derived from inorganic bases such as, for example, sodium, potassium, ammonium, calcium, or ferric hydroxides, and such organic bases as isopropylamine, trimethylamine, 2-ethylamino ethanol, histidine, procaine, and the like.

[0098] Typically, compositions are administered in a manner compatible with the dosage formulation, and in such amount as will be therapeutically effective and immunogenic. The quantity to be administered depends on the subject to be treated, including the capacity of the individual’s immune system to synthesize antibodies and the degree of protection desired. Precise amounts of active ingredient required to be administered depend on the judgment of the practitioner. However, suitable dosage ranges are of the order of several hundred micrograms active ingredient per vaccination. Suitable regimes for initial administration and booster shots are also variable, but are typified by an initial administration followed by subsequent inoculations or other administrations.

[0099] The manner of application may be varied widely. Any of the conventional methods for administration of a vaccine are applicable. These are believed to include oral application on a solid physiologically acceptable base or in a physiologically acceptable dispersion, parenterally, by injection and the like. The dosage of the vaccine will depend on the route of administration and will vary according to the size and health of the subject. [000100] The compositions and related methods, particularly administration of a peptide conjugate or nanofiber, may also be used in combination with the administration of traditional therapies. These include, but are not limited to, the administration of antibiotics such as streptomycin, ciprofloxacin, doxycycline, gentamycin, chloramphenicol, trimethoprim, sulfamethoxazole, ampicillin, tetracycline or various combinations of antibiotics. [000101] In one aspect, it is contemplated that a peptide conjugate or nanofiber or pharmaceutical composition is used in conjunction with an additional treatment described herein. Alternatively, the therapy may precede or follow the other agent treatment by intervals ranging from minutes to weeks. In embodiments where the other agents and / or a proteins is administered separately, one may generally ensure that a significant period of time did not expire between the time of each delivery, such that the agent and antigenic composition would still be able to exert an advantageously combined effect on the subject. In such instances, it is contemplated that one may administer both modalities within about 12-24 h of each other and, more preferably, within about 6-12 h of each other . In some situations, it may be desirable to extend the time period for administration significantly,Docket No.028193-0048-WO01 / 8355 however, where several days (2, 3, 4, 5, 6, or 7) to several weeks (1, 2, 3, 4, 5, 6, 7, or 8) lapse between the respective administrations. [000102] Various combinations may be employed, for example antibiotic therapy is “A” and the immunogenic composition is “B”: A / B / A B / A / B B / B / A A / A / B A / B / B B / A / A A / B / B / B B / A / B / B B / B / B / A B / B / A / B A / A / B / B A / B / A / B A / B / B / A B / B / A / A B / A / B / A B / A / A / B A / A / A / B B / A / A / A A / B / A / A A / A / B / A [000103] Administration of the pharmaceutical compositions to a patient / subject will follow general protocols for the administration of such compounds, taking into account the toxicity, if any. It is expected that the treatment cycles would be repeated as necessary. It also is contemplated that various standard therapies, such as hydration, may be applied in combination with the described therapy. [000104] In some embodiments, pharmaceutical compositions are administered to a subject. Different aspects involve administering an effective amount of a composition to a subject. In some embodiments, immunogenic compositions may be administered to the patient to protect against disease and / or inflammation. Additionally, such compounds can be administered in combination with an anti-inflammatory, a pain reliever, or other therapy. Such compositions will generally be dissolved or dispersed in a pharmaceutically acceptable carrier or aqueous medium. The use of such media and agents for pharmaceutical active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active ingredients, its use in immunogenic and therapeutic compositions is contemplated. Supplementary active ingredients can also be incorporated into the compositions. [000105] In addition to the compounds formulated for parenteral administration, such as those for intravenous or intramuscular injection, other pharmaceutically acceptable forms include, e.g., tablets or other solids for oral administration; time release capsules; and any other form currently used, including creams, lotions, mouthwashes, inhalants and the like. [000106] The active compounds can be formulated for parenteral administration, e.g., formulated for injection via the intravenous, intramuscular, sub-cutaneous, or even intraperitoneal routes. Typically, such compositions can be prepared as injectables, either as liquid solutions or suspensions; solid forms suitable for use to prepare solutions orDocket No.028193-0048-WO01 / 8355 suspensions upon the addition of a liquid prior to injection can also be prepared; and, the preparations can also be emulsified. [000107] Solutions of the active compounds as free base or pharmacologically acceptable salts can be prepared in water suitably mixed with a surfactant, such as hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms. [000108] The pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions; formulations including sesame oil, peanut oil, or aqueous propylene glycol; and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases the form must be sterile and must be fluid to the extent that it may be easily injected. It also should be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms, such as bacteria and fungi. [000109] The proteinaceous compositions may be formulated into a neutral or salt form. Pharmaceutically acceptable salts include the acid addition salts (formed with the free amino groups of the protein) and which are formed with inorganic acids such as, for example, hydrochloric or phosphoric acids, or such organic acids as acetic, oxalic, tartaric, mandelic, and the like. Salts formed with the free carboxyl groups can also be derived from inorganic bases such as, for example, sodium, potassium, ammonium, calcium, or ferric hydroxides, and such organic bases as isopropylamine, trimethylamine, histidine, procaine and the like. [000110] The carrier also can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils. The proper fluidity can be maintained, for example, by the use of a coating, such as lecithin, by the maintenance of the required particle size in the case of dispersion, and by the use of surfactants. The prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminum monostearate and gelatin.Docket No.028193-0048-WO01 / 8355 [000111] Administration of the compositions will typically be via any common route. This includes, but is not limited to oral, nasal, or buccal administration. Alternatively, administration may be by orthotopic, intradermal, subcutaneous, intramuscular, intraperitoneal, intranasal, or intravenous injection. In certain embodiments, a vaccine composition may be inhaled (e.g., U.S. Patent No.6,651,655, which is specifically incorporated by reference). Such compositions would normally be administered as pharmaceutically acceptable compositions that include physiologically acceptable carriers, buffers or other excipients. In some embodiments, the composition is administered to the subject intravenously, intraarterially, intraperitoneally, subcutaneously, intranasally, intramuscularly, or intratumorally. In some embodiments, the immunogenic composition is administered orally. In some embodiments, the immunogenic composition is administered sublingually. [000112] For parenteral administration in an aqueous solution, for example, the solution may be suitably buffered, if necessary, and the liquid diluent first rendered isotonic with sufficient saline or glucose. These particular aqueous solutions are especially suitable for intravenous, intramuscular, subcutaneous, and intraperitoneal administration. In this connection, sterile aqueous media which can be employed will be known to those of skill in the art in light of the present disclosure. For example, one dosage could be dissolved in isotonic NaCl solution and either added to hypodermoclysis fluid or injected at the proposed site of infusion, (see for example, Remington’s Pharmaceutical Sciences, 1990). Some variation in dosage will necessarily occur depending on the condition of the subject. The person responsible for administration will, in any event, determine the appropriate dose for the individual subject. [000113] An effective amount of therapeutic or prophylactic composition is determined based on the intended goal. The term “unit dose” or “dosage” refers to physically discrete units suitable for use in a subject, each unit containing a predetermined quantity of the composition calculated to produce the desired responses discussed above in association with its administration, i.e., the appropriate route and regimen. The quantity to be administered, both according to number of treatments and unit dose, depends on the protection desired. [000114] Precise amounts of the composition also depend on the judgment of the practitioner and are peculiar to each individual. Factors affecting dose include physical and clinical state of the subject, route of administration, intended goal of treatment (alleviation of symptoms versus cure), and potency, stability, and toxicity of the particular composition.Docket No.028193-0048-WO01 / 8355 [000115] Upon formulation, solutions may be administered in a manner compatible with the dosage formulation and in such amount as is therapeutically or prophylactically effective. The formulations may be easily administered in a variety of dosage forms, such as the type of injectable solutions described above. [000116] In some embodiments, the pharmaceutical compositions comprising the peptide conjugate or nanofibers detailed herein further include at least one additional therapeutic agent. The at least one additional therapeutic agent may comprise antibiotics, NSAIDS, anti-inflammatory compounds, hemoperfusion devices, quorum sensing inhibitors, lytic bacteriophage, polyclonal or monoclonal antibodies, non-immune tolerizing approaches, liposome-based cytotoxin inhibitors, or combinations thereof. The at least one additional therapeutic agent may comprise an adjuvant. Suitable adjuvants may be obtained commercially. In some embodiments, the adjuvant comprises CpG, cholera toxin B subunit (CTB), STING agonists like cyclic dinucleotides such as cyclic-di-AMP and / or cyclic GMP- AMP (cGAMP), retinoic acid, heat labile toxin B subunit, alum, MF59, 3M-052, iscomatrix, squalene-based adjuvants, AS01, AS03, or AS04, or a combination thereof. 8. Methods a. Methods of Treating an Allergy [000117] Further provided herein are methods of treating an allergy. Further provided herein are methods of reducing an allergy. The methods may include administering to a subject a conjugate peptide, or nanofiber, or pharmaceutical composition as detailed herein. The conjugate peptide, or nanofiber, or pharmaceutical composition may be administered intraperitoneally, orally, sublingually, intravenously, nasally, buccally, transdermally, intranasally, or intramuscularly. In some embodiments, the conjugate peptide, or nanofiber, or pharmaceutical composition is administered subcutaneously. [000118] Allergies may include food allergies. Food allergies may include allergies to peanuts or other legumes, eggs, nuts such as tree nuts, edible tree seeds, dairy products such as cow’s milk products, lactose, seafood, fish, shellfish, gluten, wheat, seeds such as sesame seeds and / or poppy seeds, soy, garlic, or nightshades, or a combination thereof. Tree nuts may include, for example, almonds, Brazil nuts, coconuts, cashews, chestnuts, filberts / hazelnuts, macadamia nuts, pecans, pistachios, shea nuts, pine nuts, and walnuts. Nightshades may include, for example, tomatoes, potatoes, eggplant, and peppers such as bell and chili peppers. Shellfish may include, for example, crustaceans and mollusks. Crustaceans may include, for example, shrimp, crab, and lobster. Mollusks may include, forDocket No.028193-0048-WO01 / 8355 example, mussel, oyster, scallop, squid, octopus, and snail. The compositions and methods detailed herein may reduce an allergy symptom. Allergy symptoms may include, for example, inflammation, itch, anaphylaxis, swelling such as swelling of the tongue, rash, vomiting, diarrhea, abdominal pain, hives, difficulty swallowing, hoarse voice, trouble breathing, asthma, weakened pulse, shortness of breath, trouble breathing, fainting, lightheadedness, nausea, runny nose, congested nose, or low blood pressure, or a combination thereof. [000119] In some embodiments, method further comprises administering at least one additional therapeutic agent. The at least one additional therapeutic agent may be administered prior to the conjugate peptide or the nanofiber or the pharmaceutical composition, or concurrently with the conjugate peptide or the nanofiber or the pharmaceutical composition, or after the conjugate peptide or the nanofiber or the pharmaceutical composition. The at least one additional therapeutic agent may comprise antibiotics, NSAIDS, anti-inflammatory compounds, hemoperfusion devices, quorum sensing inhibitors, lytic bacteriophage, polyclonal or monoclonal antibodies, non-immune tolerizing approaches, liposome-based cytotoxin inhibitors, or combinations thereof. The at least one additional therapeutic agent may comprise an adjuvant. In some embodiments, the adjuvant comprises CpG, cholera toxin B subunit (CTB), STING agonists like cyclic dinucleotides such as cyclic-di-AMP and / or cyclic GMP-AMP (cGAMP), retinoic acid, or heat labile toxin B subunit, or a combination thereof. [000120] The intensity of the allergy may be reduced. The duration of the allergy may be reduced. The frequency of the allergy may be reduced. A “reduced allergy” may be a reduction in intensity of the allergy, a reduction in the duration of an allergy, a reduction in the frequency of an allergy, or a combination thereof. The allergy may be reduced by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, or 10-fold, relative to a control. The allergy may be reduced by less than about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, or 10-fold, relative to a control. The allergy be reduced by about 5-95%, 10-90%, 15-85%, 20-80%, or 1.5-fold to 10-fold, relative to a control. b. Methods of Treating an Inflammatory Condition [000121] Further provided herein are methods of treating an inflammatory condition. Further provided herein are methods of treating or reducing inflammation. The methods mayDocket No.028193-0048-WO01 / 8355 include administering to a subject a conjugate peptide, or nanofiber, or pharmaceutical composition as detailed herein. The conjugate peptide, or nanofiber, or pharmaceutical composition may be administered intraperitoneally, orally, sublingually, intravenously, nasally, buccally, transdermally, intranasally, or intramuscularly. In some embodiments, the conjugate peptide, or nanofiber, or pharmaceutical composition is administered subcutaneously. [000122] In some embodiments, the inflammatory condition results from an allergy. In some embodiments, the inflammatory condition comprises inflammatory bowel disease (IBD), sepsis, ulcerative colitis, Crohn’s disease, rheumatoid arthritis, psoriasis, cirrhosis, atherosclerosis, cardiovascular inflammation, lupus, multiple sclerosis, Alzheimer’s disease, pulmonary fibrosis, immunosenescence, or a combination thereof. The inflammation may be chronic or acute. [000123] In some embodiments, method further comprises administering at least one additional therapeutic agent. The at least one additional therapeutic agent may be administered prior to the conjugate peptide or the nanofiber or the pharmaceutical composition, or concurrently with the conjugate peptide or the nanofiber or the pharmaceutical composition, or after the conjugate peptide or the nanofiber or the pharmaceutical composition. The at least one additional therapeutic agent may comprise antibiotics, NSAIDS, anti-inflammatory compounds, hemoperfusion devices, quorum sensing inhibitors, lytic bacteriophage, polyclonal or monoclonal antibodies, non-immune tolerizing approaches, liposome-based cytotoxin inhibitors, or combinations thereof. The at least one additional therapeutic agent may comprise an adjuvant. In some embodiments, the adjuvant comprises CpG, cholera toxin B subunit (CTB), STING agonists like cyclic dinucleotides such as cyclic-di-AMP and / or cyclic GMP-AMP (cGAMP), retinoic acid, or heat labile toxin B subunit, or a combination thereof. [000124] Upon or after administration, inflammation may be treated or reduced. The inflammation may be reduced by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, or 10-fold, relative to a control. The inflammation may be reduced by less than about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8- fold, 9-fold, or 10-fold, relative to a control. The inflammation be reduced by about 5-95%, 10-90%, 15-85%, 20-80%, or 1.5-fold to 10-fold, relative to a control.Docket No.028193-0048-WO01 / 8355 [000125] In some embodiments, the subject forms an antibody to the allergen epitope. Further provided herein is an antibody produced in the immune response. 9. Examples [000126] The following examples are given for the purpose of illustrating various embodiments of the invention and are not meant to limit the present invention in any fashion. One skilled in the art will appreciate readily that the present invention is well adapted to carry out the objects and obtain the ends and advantages mentioned, as well as those objects, ends and advantages inherent herein. The present examples, along with the methods described herein are presently representative of preferred embodiments, are exemplary, and are not intended as limitations on the scope of the invention. Changes therein and other uses which are encompassed within the spirit of the invention as defined by the scope of the claims will occur to those skilled in the art. Example 1 Materials and Methods 1. Examples

[0025] The foregoing may be better understood by reference to the following examples, which are presented for purposes of illustration and are not intended to limit the scope of the invention. The present disclosure has multiple aspects and embodiments, illustrated by the appended non-limiting examples.

[0026] Peptide synthesis. All peptides were synthesized using Fmoc solid-phase chemistry on a CEM Liberty Blue microwave-assisted synthesizer and purified with high- performance liquid chromatography (HPLC) and matrix-assisted laser desorption / ionization– mass spectrometry (MALDI-MS). Biotinylated peptides were synthesized on-resin by reacting Biotin-ONp (33755-53-2, Novabiochem®) with amine-terminated peptides in a threefold excess overnight in dimethyl sulfoxide (DMSO). Alexa Fluor®-647-labeled peptides were synthesized on-resin by reacting 5 mg Alexa Fluor®647 NHS Ester (Succinimidyl Ester) (ThermoFisher, Waltham, MA; A20006) with 0.05 mmol amine-terminated peptide overnight in DMSO. Peptides were cleaved for 2 hours at room temperature (i.e., from about 18°C to about 22°C) in a 95 / 2.5 / 2.5 trifluoroacetic acid / triisopropylsilane / water cocktail, followed by washing with cold diethyl ether. Peptides were purified by reversed-phase high- performance liquid chromatography using a C18 column and lyophilized. TABLE 1 shows a complete list of peptides that were used.Docket No.028193-0048-WO01 / 8355* Peanut peptide epitopes from the major peanut allergens Ara h 1, 2, 3, and 7 were selected from the literature for both broad-reactivity and immunodominance. Sources from which each peptide epitope was selected are provided. Ac: Acetyl; Am: Amide; X: cyclohexylalanine; a: D-alanine.

[0027] Nanofiber vaccine formulation. Q11 nanofibers were formed as previously described (Shores et al. Frontiers in Immunology 2020, 11, 1-13; Solano, et al. (2018)Docket No.028193-0048-WO01 / 8355 Practical Considerations in the Design and Use of Immunologically Active Fibrillar Peptide Assemblies. In: Nilsson, B., Doran, T. (eds) Peptide Self-Assembly. Methods in Molecular Biology, vol 1777. Humana Press, New York, NY; Mora-Solano et al. Biomaterials 2017, 149, 1-11; each of which is incorporated herein by reference). Briefly, lyophilized peptides were dissolved at 8 mM in sterile water and incubated at 4°C overnight. The solutions were then brought to a final peptide concentration of 2mM in 1× phosphate-buffered saline (PBS) by addition of sterile water and sterile 10× PBS and incubated at room temperature for 3 hours before use to allow for fibrillization. Unless otherwise specified, (OVA61-68)Q11 nanofibers were formulated with 25% (OVA61-68)Q11 peptide content. All peanut epitope nanofiber formulations were formulated with 25% (Epitope)Q11 peptide content. Nanofiber formulations used in immunization experiments included 2.5% PADRE-Q11 peptide to provide a helper T cell epitope. For adjuvanted formulations, CpG (InvivoGen, San Diego, CA; tlrl-1826) was added just before fibrillization corresponding to a concentration of 100 μg / mL, or a dose of 10 μg of CpG per mouse.

[0028] Transmission electron microscopy (TEM). To visualize nanofiber morphology by transmission electron microscopy, nanofiber solutions were diluted to 0.2 mM in 1× PBS and deposited onto Formvar® / carbon-coated 400 mesh copper grids (Electron Microscopy Sciences, Hatfield, PA; EMS400-Cu) for 1 min, rinsed with ultrapure water, and negatively stained for 1 min with 1% (w / v) uranyl acetate (EMS, 22400-1) before wicking away with filter paper. Samples were imaged on an FEI Tecnai G2 Twin®electron microscope at 120 kV.

[0029] Model protein allergen formulation. To formulate model protein allergen, equal masses of biotinylated OVA61-68peptide and purified streptavidin (Biolegend, San Diego, CA; 405151) were incubated at a 1 mg / mL streptavidin concentration in PBS. Excess unbound peptide was removed via centrifugal filtration using 10 kDa cutoff Amicon™ Ultra-0.5 Centrifugal Filter Units (MilliporeSigma, Burlington, MA; UFC501096) according to the manufacturer’s instructions.

[0030] Animal experiments. Animal experiments were performed using 8-12 week old age- and sex-matched BALB / c (Strain cAnNHsd) female mice purchased from Envigo (Indianapolis, IN) and housed at the animal facility of Duke University (Durham, NC). All animal procedures were performed in accordance with and approved by the Institutional Animal Care and Use Committee of Duke University under protocol #A199-21-09.

[0031] Immunizations. Mice were anesthetized under isoflurane and immunized subcutaneously at the tail base with indicated solutions with 50 L at each side of the tail base (for nanofiber vaccines, 200 nmol each peptide epitope per mouse). At indicated timeDocket No.028193-0048-WO01 / 8355 points, blood samples were collected from submandibular vein to analyze for allergen- specific sera antibodies via ELISA. To purify IgG for subsequent analysis in BMMC activation assays, serum was pooled, and 0.2 mL NAb™ Protein G Spin columns were used according to the manufacturer’s instructions. After IgG purification, the resulting purified IgG was buffer-exchanged to a balanced salt solution consisting of 135 mM NaCl, 12.5 mM MgCl2, 1.8 mM CaCl2, 20 mM HEPES, and 5.6 mM Glucose using 10 kDa cutoff Amicon®Ultra-0.5 Centrifugal Filter Units (MilliporeSigma, Burlington, MA; UFC501096) according to the manufacturer’s instructions.

[0032] Systemic vaccine exposure. Mice were immunized as described above with the indicated Alexa Fluor®647-labeled vaccines. Small blood samples (~10 μL) were collected via tail snip into heparinized capillary tubes. Blood was removed from capillary tubes and diluted 1:1.5 in 1x PBS. Samples were centrifuged at 600 rcf for 5 minutes to pellet cells. Supernatant was removed and transferred to a new tube. Fluorescence was measured on a Nanodrop 3300 (ThermoFisher, Waltham, MA) at an emission of 670 nm, values reported as relative fluorescence units (RFU).

[0033] Passive sensitization and allergen challenge. Mice were administered the indicated amount (either 50 or 55 g) of E-C1 IgE mAb (Chondrex, Woodinville, WA; 3006) in 100 μL of sterile PBS via tail vein injection. The next day, mice were challenged with 100 μL of the indicated allergen solutions via the indicated route (subcutaneous at the tail base or intraperitoneally). To monitor systemic allergic responses, body temperature was measured via rectal thermometer and serum was collected immediately before and 1 hour after challenge for analysis of mast cell protease 1 (MCPT-1) levels. Significant hypothermia (rectal temperatures below 32 °C) was considered a humane endpoint. Serum levels of MCPT-1 were analyzed using a MCPT-1 (mMCP-1) Mouse Uncoated ELISA Kit (ThermoFisher, Waltham, MA; 88-7503-22) according to manufacturer’s instructions.

[0034] Active allergen sensitization. For sensitization against crude peanut extract, mice were injected intraperitoneally with 100 g of crude peanut extract (Stallergenes Greer, Baar, Switzerland; F171 peanut source material, XPF171D3A2.5, Lot# 352557) mixed with 200 μL of AlHydrogel®adjuvant (InvivoGen, San Diego, CA; vac-alu-250) once a week for a total of 3 weeks. Serum was collected, pooled, and depleted of IgG using 1 mL NAb™ Protein G Spin columns (ThermoFisher, Waltham, MA; 89979). The IgG-depleted serum was then used to sensitize BMMC (see BMMC activation assays section below).

[0035] Peritoneal mast cell activation assay. Mice were sensitized to OVA61-68via intraperitoneal injection of 25 μg E-C1 IgE mAb (Chondrex, Woodinville, WA; 3006) in 250Docket No.028193-0048-WO01 / 8355 μL of sterile PBS. The next day, 500 μL of a 0.2 mM (OVA61-68)Q11 nanofiber solution or an 11 μg / mL OVA61-68tetramer solution was injected intraperitoneally. One hour later, mice were sacrificed, and a lavage of the peritoneal cavity was performed with 500 μL of sterile PBS + 1% bovine serum albumin. Lavage fluid was filtered through a 70 μm cell strainer and centrifuged to pellet cells. Peritoneal mast cell activation (CD63 surface expression) was analyzed via flow cytometry. See FIG.2 for flow cytometry gating strategy.

[0036] Enzyme-linked immunosorbent assays (ELISA). For analysis of peptide epitope- specific antibody by ELISA, plates were coated with 20 g / mL streptavidin (MilliporeSigma, Burlington, MA; 189730) overnight at 4°C. Plates were washed with Tween 20 (0.5 g / liter) in PBS (1x PBST). Biotinylated peptides (20 g / mL) were captured for 30 minutes. For analysis of protein-specific antibody, ELISA plates were coated with 5 μg / mL crude peanut extract (Stallergenes Greer, Baar, Switzerland; F171 peanut source material, XPF171D3A2.5, Lot# 352557), natural Ara h 1 (InBio, Charlottesville, VA; NA-AH1-1), natural Ara h 2 (InBio, Charlottesville, VA; NA-AH2-1), natural Ara h 3 (InBio, Charlottesville, VA; NA-AH3-1), or recombinant Ara h 7 (RayBiotech, Peachtree Corners, GA; 230-00901- 50). Coated plates were blocked with Super Block Blocking Buffer (ThermoFisher Scientific, Waltham, MA; 37515). Sera diluted in 1x PBST + 1% bovine serum albumin were added to the plate for 1.5 hours and plates were washed in 1x PBST. To detect antigen-specific IgG,horseradish peroxidase (HRP)–conjugated Fc fragment–specific goat anti-mouse IgG(Jackson ImmunoResearch, West Grove, PA; 15-035-071) was added to the plates. To detect antigen-specific IgE, HRP-conjugated goat anti-mouse IgE (Southern Biotech, Birmingham, AL; 1110-05) was added to the plates. Plates were washed in 1x PBST and TMB substrate (Invitrogen, Waltham, MA; 00-4201-56) was added to the plates for 5 minutes. Development was stopped with 1M H3PO4and absorbance was read at 450 nm on a microplate reader. Results are reported as antibody titers calculated with an absorbance cutoff of 0.2 OD (optical density), or as background subtracted A450 (absorbance at 450 nm) values. Samples below the limit of detection were assigned a titer of zero.

[0037] BMMC culture. Bone marrow was isolated from mouse femurs and tibias and passed through a 70 μm cell strainer. Red blood cells were lysed using ACK cell lysis buffer (Gibco, Waltham, MA; A10492-01) and the remaining cells were cultured on tissue culture- treated cell culture flasks in BMMC culture media [RPMI 1640 with l-glutamine, 10 mM Hepes, 50 M -mercaptoethanol, 1 mM sodium pyruvate, 1 mM nonessential amino acids, and 10% fetal bovine serum (FBS) + Penicillin-Streptomycin] supplemented with 10 ng / mL recombinant mouse interleukin 3 (rmIL-3) (R&D Systems, Minneapolis, MN; 403-ML). Every 3-4 days, non-adherent cells were pelleted via centrifugation, resuspended in fresh BMMCDocket No.028193-0048-WO01 / 8355 culture media + 10 ng / mL rmIL-3, and moved to a new tissue culture flask. All BMMC experiments were performed with cells cultured for 4 to 8 weeks.

[0038] BMMC activation assays. For assays of BMMC activation in response to the OVA61-68model allergen epitope, BMMC were sensitized for 16 hours with 0.1 μg / mL E-C1 IgE mAb (Chondrex, Woodinville, WA; 3006) at a cell density of 500K / mL in BMMC culture media. BMMC were washed to remove excess IgE and plated at 500K / mL and stimulated with the indicated solutions. For assessment of sIgG-mediated allergen neutralization, 400 g / mL purified IgG was pre-mixed with allergen for 30 minutes prior to addition to BMMC. BMMC were stimulated for 45 minutes before analysis of activation via flow cytometry. See FIG.3 for flow cytometry gating strategy.

[0039] For assays of BMMC activation in response to peanut allergen proteins, BMMC were sensitized via 24-hour incubation in 10% IgG-depleted allergic serum (see active allergen sensitization section) in BMMC culture media supplemented with 10 ng / mL rmIL-3 (R&D Systems, Minneapolis, MN; 403-ML) at a cell density of 500K / mL. BMMC were then washed and plated in BMMC culture media overnight. BMMC were washed and plated at a cell density of 500K / mL with the indicated allergen solutions at a concentration of 10 ug / mL allergen. For assessment of sIgG-mediated allergen neutralization, 400 g / mL purified IgG was pre-mixed with allergen for 30 minutes prior to addition to BMMC. BMMC were stimulated for 45 minutes before analysis of activation via flow cytometry.

[0040] To quantify BMMC activation, cells were stained for CD45, Fc RI, c-kit, andCD63 in FACS buffer (1x PBS + 1% bovine serum albumin + 1 mM EDTA) using FITC anti-mouse CD45 (Biolegend, San Diego, CA; 103108), PE anti-mouse Fc RI (Biolegend, SanDiego, CA; 134308), Brilliant Violet 510™ anti-mouse CD117 (c-kit) (Biolegend, San Diego, CA; 105839), and APC anti-mouse CD63 (Biolegend, San Diego, CA; 143906). DAPI wasincluded as a viability dye at 1 μg / mL. BMMC were defined as viable CD45+Fc RI+cKIT+cells and activation was quantified via CD63 expression (MFI, mean fluorescent intensity). CD63 MFI of unstimulated cells was subtracted as background signal.

[0041] Human peanut allergic serum. Serum from peanut allergic human donors was obtained from PlasmaLab International (Everett, WA; Donor codes: 33599-KA, 32342-DG, 26730-AB, 25373-AD, 33454-PS, 34480-NV, 29927-MF, 20197-BH).

[0042] Humanized RBL-NFAT-DsRed culture. Parental RBL-NFAT-DsRed cells (Ali etal., Plos one 2019; 14 (8): e0221034) expressing the human Fc RI and Fc RI chains wereobtained from the University of Nottingham, Nottingham, England. Cells were maintained inDocket No.028193-0048-WO01 / 8355 RBL Culture Media (MEM with GlutaMAX™ Supplement + 10% FBS + Penicillin- Streptomycin). Every 3-4 passages, cells were treated with 1 mg / mL G418 sulfate, 20 μg / mL Blasticidin S.

[0043] Humanized RBL-NFAT-DsRed sorting. To improve the sensitivity of the assays herein, functional reporter cells were sorted from the parental humanized RBL-NFAT-DsRed cell line. Cells were plated in a 24 well plate at 7.5 x 104cells per well and incubated overnight. The next day cells were sensitized with 0.2 m sterile filtered 2.5% allergic human serum in RBL culture media. Prior to dilution in RBL culture media, allergic serum was heat inactivated via incubation at 56°C for 5 minutes. The next day, cells were washed and stimulated with 1 μg / mL goat anti-human IgE for 24 hours. Cells were washed, trypsinized, suspended in FACS buffer and DsRed+ cells were sorted using a Sony MA900 cytometer. The sorted cell population was diluted in RBL culture media to ~1 cell / 100 μL and plated in a 96-well plate to isolate and expand single cell clones. A single cell clonal line was selected for expansion and used for subsequent experiments.

[0044] Humanized RBL-NFAT-DsRed activation assay. RBL-NFAT-DsRed cells were plated in a 96-well plate at 1.25 x 104cells per well and incubated overnight. The next day, cells were sensitized for 24 hours with 0.2 m sterile filtered serum from peanut allergic donors at 5% allergic serum in RBL culture media. Prior to dilution in RBL culture media, allergic serum was heat inactivated via incubation at 56°C for 5 minutes. The following day, cells were washed and treated overnight with solutions of 0.05 mM peanut epitope nanofibers or Q11-only nanofibers, or with 1 g / mL crude peanut extract as a positive control (Stallergenes Greer, Baar, Switzerland; F171 peanut source material, XPF171D3A2.5, Lot# 352557). Cells were washed, trypsinized, and suspended in FACS buffer for flow cytometry analysis of DsRed expression. DAPI was included as a viability dye. Flow cytometry was performed using BD Canto II instrument and analyzed using FlowJo software. Cells sensitized with serum from donor 20197-BH failed to respond to the positive control and were excluded from the analysis.

[0045] SST design and assembly. DNA strands were synthesized by Integrated DNA Technology (idtdna.com) (TABLE 2 and TABLE 3). The 6-helix-bundle SST DNA nanostructure was designed with the software caDNAno (cadnano.org). To assemble the structures, unpurified DNA strands were mixed in an equimolar stoichiometric ratio from a 100 μM stock in 1x Tris / EDTA buffer (10 mM Tris, 1 mM EDTA, pH 8.0) supplemented with 12.5 mM MgCl2. 6HB with or without DNP, was annealed in a PCR thermocycler by heating at 95 °C for 3 mins and slowly cooling from 65 to 25 °C at a rate of -0.1 °C / 3 min. The foldedDNA nanostructures were stored at 4 °C.Docket No.028193-0048-WO01 / 8355Docket No.028193-0048-WO01 / 8355Docket No.028193-0048-WO01 / 8355Docket No.028193-0048-WO01 / 8355Docket No.028193-0048-WO01 / 8355

[0046] Agarose gel electrophoresis analysis. DNA nanostructures samples were analyzed using agarose gel electrophoresis with 2.5% agarose gel pre-stained with ethidium bromide. Samples were run at 75 V for 2-3 hours in an ice water bath and visualized with a gel imager (Bio-Rad, Hercules, CA).

[0047] Streptavidin labelling. To visualize 6HB with different spacing of overhangs, DNA nanostructures are labeled with streptavidin via biotin-streptavidin binding. Biotin modified sequences were mixed with 4x amounts of streptavidin and left to react for 2h at room temperature. Then 6HB with corresponding overhangs were incubated with 10x excess of biotin-streptavidin for 2h at room temperature. Streptavidin labeled 6HB were purified by 2% agarose gel electrophoresis. The migrating bands corresponding to the correct structures were then cut out from the gel. Excised bands were crushed and transferred into DNA gel extraction spin column (Bio-Rad, Hercules, CA; catalogue number: 7326165). The DNAstructure solution was recovered by centrifugation of the loaded column for 10 min at16,000 g.

[0048] Atom Force Microscopy (AFM). Two microliters of the solution were deposited onto freshly cleaved mica. The sample area was then washed by water twice and dried by nitrogen gas. The samples were imaged with a Multimode VIII system (Bruker, Billerica, MA) in air using commercial tips (SCANASYST®-AIR, Bruker, Billerica, MA).

[0049] Assessing mast cell responses to SST model immunogens.

[0050] BMMC culture. Bone marrow was isolated from mouse femurs and tibias and passed through a 70 μm cell strainer. Red blood cells were lysed using ACK cell lysis bufferDocket No.028193-0048-WO01 / 8355 (Gibco, Waltham, MA; A10492-01) and the remaining cells were cultured on tissue culture- treated cell culture flasks in BMMC culture media [RPMI 1640 with l-glutamine, 10 mM HEPES, 50 M -mercaptoethanol, 1 mM sodium pyruvate, 1 mM nonessential amino acids, and 10% fetal bovine serum (FBS) + Penicillin-Streptomycin] supplemented with 10 ng / mL recombinant mouse interleukin 3 (rmIL-3, R&D Systems, Minneapolis, MN; 403-ML). Every 3-4 days, non-adherent cells were pelleted via centrifugation, resuspended in fresh BMMC culture media + 10 ng / mL rmIL-3, and moved to a new tissue culture flask. All BMMC experiments were performed with cells cultured for 4 to 8 weeks.

[0051] SST-induced bone-marrow derived mast cell (BMMC) activation assay. BMMC were sensitized with 0.1 μg / mL SPE-7 IgE (D8406, Millipore Sigma, Burlington, MA) for 16 hours at 0.5 x 106cells / mL in BMMC culture media. BMMC were washed to remove excess IgE and stimulated at 0.5 x 106cells / mL with the indicated SST model immunogens for 45 minutes. Stimulation was carried out in stimulation buffer (135 mM NaCl, 12.5 mM MgCl2, 1.8 mM CaCl2, 20 mM HEPES, 5.6 mM Glucose + 1% nuclease-free BSA (Millipore Sigma, Burlington, MA; 126609), pH 7.4). To quantify BMMC activation, cells were stained forCD45, Fc RI, c-kit, and CD63 in FACS buffer (1x PBS + 1% bovine serum albumin + 1 mMEDTA) using FITC anti-mouse CD45 (Biolegend, San Diego, CA; 103108), PE anti-mouseFc RI (Biolegend, San Diego, CA; 134308), Brilliant Violet 510™ anti-mouse CD117 (c-kit)(Biolegend, San Diego, CA; 105839), and APC anti-mouse CD63 (Biolegend, San Diego, CA; 143906). DAPI was included as a viability dye at 1 μg / mL. BMMC were defined asviable CD45+Fc RI+cKIT+ cells and activation was quantified via CD63 expression (MFI,mean fluorescent intensity). CD63 MFI of unstimulated cells was subtracted as background signal. Negative values were defined as zero.

[0052] SST-induced-peritoneal mast cell activation assay. Mice were sensitized to DNP via intraperitoneal injection of 10 μg SPE-7 IgE in 250 μL of PBS. The next day, mice were challenged intraperitoneally with the indicated SST model immunogens at 20 nM DNP concentration in 500 μL of stimulation buffer. One hour later, mice were sacrificed, and a lavage of the peritoneal cavity was performed with 500 μL of sterile PBS + 1% bovine serum albumin. Lavage fluid was filtered through a 70 μm cell strainer and centrifuged to pelletcells. Cells were stained for CD45, Fc RI, c-kit, and CD63 in FACS buffer (1x PBS + 1%bovine serum albumin + 1 mM EDTA) using FITC anti-mouse CD45 (Biolegend, San Diego,CA; 103108), PE anti-mouse Fc RI (Biolegend, San Diego, CA; 134308), Brilliant Violet510™ anti-mouse CD117 (c-kit) (Biolegend, San Diego, CA; 105839), and APC anti-mouse CD63 (Biolegend, San Diego, CA; 143906). DAPI was included as a viability dye at 1Docket No.028193-0048-WO01 / 8355μg / mL. Peritoneal mast cells (PMC) were defined as viable CD45+Fc RI+cKIT+ cells andactivation was quantified via CD63 expression (MFI, mean fluorescent intensity).

[0053] RAMOSDNP B cell model.

[0054] SPE-7 tgBCR gene constructs. To construct chimeric anti-DNP B cell receptor (BCR) genes, the heavy and light chain variable region sequences of the murine monoclonal IgE clone SPE-7 (PDB: 1OAU) were appended to the constant region sequences of human membrane-bound IgM (Uniprot ID: P01871-2) and lambda light chain (Uniprot ID: P0DOX8), respectively. A mammalian antibody heavy and light chain coexpression PiggyBac Vector was constructed by VectorBuilder®in which both heavy and light chain genes are flanked by PiggyBac ITR sequences. A pRP[Exp]-CMV>HA / hyPBase expression vector encoding the PiggyBac transposase was also obtained from VectorBuilder®.

[0055] RAMOSDNP B cell engineering. RAMOS B cells were cultured in RPMI supplemented with 15% heat-inactivated fetal bovine serum (FBS) and Penicillin- Streptomycin. First, BCR negative cells were isolated and expanded from the parental RAMOS cell line. Cells were stained with Alexa Fluor®488 anti-human IgM (Biolegend, San Diego, CA; 314533) and PE Mouse Anti-Human Light Chain, (BD Biosciences, Franklin Lakes, NJ; 562054) and BCR negative cells were sorted using a Beckman Coulter Astrios®cell sorter. LIVE / DEAD™ Fixable Violet (Thermofisher, Waltham, MA; L34964) was included as a viability dye. BCR-negative cells were expanded, and an additional round of sorting was performed to further enrich BCR negative cells.

[0056] 1 x 106BCR negative RAMOS B cells were suspended in 100 μL of Opti-MEM™ I Reduced Serum Medium (Thermofisher, Waltham, MA; 31985062) with 5 μg of the SPE-7 IgM BCR expression vector and 3 μg of the pRP[Exp]-CMV>HA / hyPBase expression vector. Cells were transferred to 4 mm gap electroporation cuvettes and electroporated with two 10 ms pulses of 250 V spaced 10 s apart. After electroporation, cells were transferred directly to culture media and expanded. On day 9 of culture after electroporation, BCR positive cells were selected using a Beckman Coulter Astrios®cell sorter. SPE-7 IgM BCR-expressing cells were identified using Alexa Fluor®488 anti-human IgM (Biolegend, San Diego, CA; 314533) and DNP-BSA (Millipore Sigma, Burlington, MA; 324101) which was then detected with polyclonal goat anti-DNP followed by Alexa Fluor®647 anti-goat IgG. LIVE / DEAD™ Fixable Violet (Thermofisher, Waltham, MA; L34964) was included as a viability dye. IgM+ DNP-BSA+ cells were isolated and expanded for 7 days before an additional round of sorting was performed to ensure a pure population of SPE-7 IgM BCR-expressing RAMOS B cells (RAMOSDNP). RAMOSDNP cells were expanded and BCR expression was validatedDocket No.028193-0048-WO01 / 8355 using an identical cell staining procedure with DAPI included as a viability dye. Samples were run on a BD Canto flow cytometer.

[0057] RAMOSDNP B cell BCR signaling validation using calcium flux assay. Functional validation of BCR signaling in RAMOSDNP cells was performed using calcium flux assay. RAMOSDNP cells were incubated in 5 μM Fluo-4, AM (Thermofisher, Waltham, MA; F14201) + 2.5 mM water-soluble probenecid (Thermofisher, Waltham, MA; P36400) in serum free RPMI media at 1 x 106cells / mL for 30 min at 37°C. Cells were washed with serum free RPMI media and resuspended at 1 x 106cells / mL and incubated for 30 min at 37°C. Cells were then washed again and resuspended in stimulation buffer (135 mM NaCl, 12.5 mM MgCl2, 1.8 mM CaCl2, 20 mM HEPES, 5.6 mM Glucose + 1% nuclease-free BSA (Millipore Sigma, Burlington, MA; 126609), pH 7.4) + 2.5 mM probenecid. Cells were maintained at 37 °C and 300 μL were transferred to pre-warmed tubes immediately prior to running on a BD Canto flow cytometer. Baseline Fluo-4 signal was collected for at least 10 seconds prior to the addition of DNP-BSA. Ionomycin (Thermofisher, Waltham, MA; 124222) was used as a positive control. Fluo-4 signal was measured for at least 90 seconds after stimulation. All flow cytometry data was processed and analyzed using FlowJo software.

[0058] Assessing B cell responses to SST model immunogens.

[0059] SST-induced pERK expression assay. RAMOSDNP cells were suspended at 1 x 106cells / mL in stimulation buffer and stimulated with SST model immunogens or DNP-BSA at 37°C for the indicated amount of time. Cells were immediately cooled on ice and stained with 1:200 LIVE / DEAD™ Fixable Violet (Thermofisher, Waltham, MA; L34964) in FACS buffer (1x PBS + 1% bovine serum albumin + 1 mM EDTA) for 10 min. Cells were washed with FACS buffer and fixed in 2% paraformaldehyde in PBS for 10 minutes at room temperature. Cells were washed with PBS and permeabilized with ice cold 90% methanol for 30 minutes on ice. Cells were washed with PBS and labeled with Phospho-p44 / 42 MAPK (Erk1 / 2) (Thr202 / Tyr204) (197G2) Rabbit mAb #4377 (Cell Signaling Technologies, Danvers, MA; 4377S) diluted 1:80 in FACS buffer for 45 minutes at room temperature. Cellswere washed and stained with Allophycocyanin (APC) AffiniPure F(ab') Fragment DonkeyAnti-Rabbit IgG (H+L) (Jackson Immunoresearch, West Grove, PA; 711-136-152) before running on a BD Canto Flow cytometer. For each experimental replicate, pERK values were normalized using the positive control group DNP-BSA as a maximum (set to 1.0) and the negative control group 6HB as a minimum (set to 0.0).Docket No.028193-0048-WO01 / 8355

[0060] SST-induced calcium flux assay. RAMOSDNP cells were processed and labeled with Fluo-4 and ran on a BD Canto flow cytometer as described above. Concentrated SST immunogens (1 μM 6HB scaffold) were added directly to cells after acquisition of baseline Fluo-4 signal yielding a final concentration of 20 nM DNP in each sample. Samples were briefly vortexed, and Fluo-4 signal was measured immediately over a period of at least 90 seconds. Three technical replicates were completed, with each replicate containing a DNP- BSA positive control and a 6HB (no DNP SST) negative control. Baseline signal was removed from each sample, and within each replicate set, signal was normalized using the positive control group DNP-BSA as a maximum (set to 1.0) and the negative control group 6HB as a minimum (set to 0.0). AUC values were calculated for each curve, and again normalized in the same fashion. To analyze distribution and magnitude of B cell activation at baseline and during stimulation, gates were drawn to select cells at baseline and during the peak of the Fluo-4 signal for each sample. Within the peak gate, cells were gated once more to distinguish activated cells from non-activated cells.

[0061] Validation of tunable control of nanofiber epitope density through co-assembly.

[0062] Peptide synthesis. All peptides were synthesized using Fmoc solid-phase chemistry on a CEM Liberty Blue microwave-assisted synthesizer and purified with high- performance liquid chromatography (HPLC) and matrix-assisted laser desorption / ionization– mass spectrometry (MALDI-MS). Biotinylated peptides were synthesized on-resin by reacting Biotin-ONp (33755-53-2, Novabiochem, London, GB) with amine-terminated peptides in a threefold excess overnight in dimethyl sulfoxide (DMSO). All peptide conjugates were synthesized via on-resin N-terminal conjugation. Biotin-ONp (33755-53-2, Novabiochem, London, GB) was conjugated using p-nitrophenyl ester coupling. DNP- dPEG®-NHS ester (10347, Quanta Biodesign, Union County, OH) and NHS-Fluorescein (46409, Thermofisher, Waltham, MA) were conjugated using N-hydroxysuccinimide ester coupling. 5(6)-TAMRA (AS-81120-01, AnaSpec, Fremont, CA) was conjugated using 1- ethyl-3-(3-(dimethylamino)propyl)carbodiimide coupling. Peptides were cleaved off of resin for 2 hours at room temperature in a 95 / 2.5 / 2.5 trifluoroacetic acid / triisopropylsilane / water cocktail, followed by washing with cold diethyl ether. Peptides were purified by reversed- phase high-performance liquid chromatography using a C18 column and lyophilized. See TABLE 4 for a complete list of peptides used in this study.Docket No.028193-0048-WO01 / 8355Ac: Acetyl; Am: Amide. Bold text indicates a peptide conjugate. Italic text indicates linker sequence.

[0063] Nanofiber formation. Individual peptides were dissolved in DMSO at 40 mM before mixing at the indicated molar ratios. Peptide solutions were brought up to 8 mM 95°C acetate buffer (10 mM Sodium acetate, pH 4) and then brought to 2 mM with 95°C water. Solutions were heated at 95°C for 15 minutes and then incubated at room temperature overnight.

[0064] Quantification of nanofiber fluorescence and FRET activity. Nanofiber solutions were diluted 1:1000 in acetate buffer and 10 μL was pipetted onto a glass slide and topped with a #1.5 cover slip. Samples were imaged on a Leica SP8 inverted confocal microscope at 100X magnification. All images were processed and analyzed in ImageJ. Images were binarized via the “Adjust threshold” function and the “Analyze particles” function was used to create region of interest (ROI) masks to identify individual nanofibers. Aggregated nanofibers were excluded from the analysis by limiting ROI size. The mean grey value for each nanofiber ROI was measured in each image channel. For FRET experiments, nanofiber intensity values were adjusted for crosstalk between image channels via compensation. Briefly, the crosstalk of TAMRA-only or FITC-only nanofiber signal into other image channels was calculated as a fraction of the TAMRA or FITC signal, respectively, and the average crosstalk of each was subtracted from each image channel. Thus, the contribution of TAMRA signal into the FITC and FRET channels, and the contribution of FITC into the TAMRA and FRET channels was removed.

[0065] Assessing mast cell and B cell responses to Coil29 nanofiber immunogens.

[0066] Nanofiber Atomic Force Microscopy (AFM). (OVA61-68)C29 nanofibers with varying epitope densities were formed as described above for AFM imaging as described elsewhere (Fries et al., Advanced Materials 2020; 32: 2003310). Briefly, nanofibers were diluted to 0.2 mM in water. Mica substrates (TedPella, Inc., Redding, CA; Cat #50) wereDocket No.028193-0048-WO01 / 8355 cleaved immediately prior to sample preparation, and 20 L of the diluted nanofiber solution was pipetted onto the surface. After 30 seconds, substrates were rinsed with ultrapure water and dried with nitrogen gas. Imaging was conducted using a Bruker AFM on tapping mode with RTESPA-300 silicon tips.

[0067] Nanofiber-induced BMMC activation assay. BMMC were cultured as described above. BMMC were sensitized with either 0.1 μg / mL SPE-7 IgE (D8406, Millipore Sigma, Burlington, MA) or 0.1 μg / mL E-C1 IgE (Chondrex, Woodinville, WA; 3006) for 16 hours at 0.5 x 106cells / mL in BMMC culture media. Cells were washed and stimulated with the indicated nanofiber formulations at the indicated concentrations in BMMC media for 45 minutes at 37°C. Quantification of CD63 surface expression via flow cytometry was performed as described above.

[0068] Nanofiber-induced RAMOSDNP B cell pERK expression assay. RAMOSDNP cells were stimulated for 5 minutes with the indicated nanofiber formulations in culture media at a concentration of 100 nM DNP (5% DNP nanofibers diluted to 2 μM total peptide; 0.5% nanofibers diluted to 20 μM total peptide; Coil29-only nanofibers were matched to the higher 20 μM total peptide concentration). pERK expression was quantified via flow cytometry as described above.

[0069] Assessing the therapeutic efficacy of low-density nanofiber allergen vaccine immunogens in a passive sensitization model of allergic anaphylaxis.

[0070] Animals. Animal experiments were performed using 8-12 week old age- and sex- matched BALB / c (Strain cAnNHsd) female mice purchased from Envigo (Indianapolis, IN) and housed at the animal facility of Duke University (Raleigh, NC). All animal procedures were performed in accordance with and approved by the Institutional Animal Care and Use Committee of Duke University under protocol #A199-21-09.

[0071] Adoptive transfer of splenocytes from allergen-sensitized mice. OVA61-68was synthesized with a SGSG-C-SGSG (SEQ ID NO: 105) C-terminal linker and conjugated to Imject™ Maleimide PEGylated mcKLH (Thermofisher, Waltham, MA; 77663) according to the manufacturer’s instructions. OVA61-68-KLH was diluted to 1 mg / mL and mixed 1:1 with Alhydrogel®adjuvant 2% (InvivoGen, San Diego, CA; vac-alu-50). Mice were sensitized via three weekly intraperitoneal injections of 200 μL. Three weeks after the last immunization, mice were sacrificed and splenocytes were isolated and pooled. 20 x 106cells were transferred to naïve mice via intraperitoneal injection.Docket No.028193-0048-WO01 / 8355

[0072] Nanofiber immunization. 2 mM nanofiber solutions were brought to physiological osmolarity via addition of 10X PBS prior to immunization. Mice were immunized intraperitoneally with 100 μL of nanofiber solution at weeks 0, 3, and 5. At indicated time points, blood samples were collected from submandibular vein to analyze for allergen- specific sera antibodies via ELISA.

[0073] ELISA. For analysis of peptide epitope-specific antibody by ELISA, plates were coated with 20 g / ml streptavidin (Millipore Sigma, Burlington, MA; 189730) overnight at 4°C. Plates were washed with Tween 20 (0.5 g / liter) in PBS (1x PBST). Biotinylated peptides (20 g / mL) were captured for 30 minutes. Sera diluted in 1x PBST + 1% bovine serum albumin were added to the plate for 1.5 hours and plates were washed in 1x PBST.To detect antigen-specific IgG, horseradish peroxidase (HRP)–conjugated Fc fragment–specific goat anti-mouse IgG (Jackson ImmunoResearch, West Grove, PA; 15-035-071) was added to the plates. Plates were washed in 1x PBST and TMB substrate (Invitrogen, Waltham, MA; 00-4201-56) was added to the plates for 5 minutes. Development was stopped with 1M H3PO4and absorbance was read at 450 nm on a microplate reader. Results are reported as area under the curve (AUC) calculated using background-subtracted absorbance values.

[0074] Allergen neutralization BMMC assay. To purify IgG for subsequent analysis in BMMC activation assays, serum was pooled, and 0.2 mL NAb™ Protein G Spin columns were used according to the manufacturer’s instructions. After IgG purification, the resulting purified IgG was buffer-exchanged to stimulation buffer using 10 kDa cutoff Amicon™ Ultra- 0.5 Centrifugal Filter Units (Millipore Sigma, Burlington, MA; UFC501096) according to the manufacturer’s instructions.

[0075] Because the OVA protein contains only one copy of the OVA61-68epitope, andmast cells require multivalent antigen to activate through IgE-Fc RI, model allergentetramers were used to stimulate BMMC and challenge passively sensitized mice. To formulate model protein allergen, equal masses of biotinylated OVA61-68peptide (~20-fold molar excess) and purified streptavidin (Biolegend, San Diego, CA; 405151) were incubated at a 1 mg / mL streptavidin concentration in PBS. Excess unbound peptide was removed via centrifugal filtration using 10 kDa cutoff Amicon™ Ultra-0.5 Centrifugal Filter Units (Millipore Sigma, Burlington, MA; UFC501096) according to the manufacturer’s instructions.

[0076] BMMC were cultured as described above. Cells were sensitized with 1 μg / mL E- C1 IgE in BMMC culture media for 16 hours before stimulation with 1 μg / mL allergen tetramer and 500 μg / mL purified IgG. Prior to stimulation, model allergen tetramers wereDocket No.028193-0048-WO01 / 8355 pre-incubated with the indicated IgG for 30 min. Cells were stimulated for 45 minutes at 37°C and BMMC activation was quantified as described above.

[0077] Passive sensitization and allergen challenge. On week 6 (one-week after the last booster immunization), mice were passively sensitized to the OVA61-68model allergen via intravenous injection of 55 μg E-C1 IgE. The next day, mice were challenged with 5 μg of model allergen tetramer via intraperitoneal injection. To monitor systemic allergic responses, body temperature was measured via rectal thermometer and serum was collected immediately before and 1 hour after challenge for analysis of mast cell protease 1 (MCPT-1) levels. Serum levels of MCPT-1 were analyzed using a MCPT-1 (mMCP-1) Mouse Uncoated ELISA Kit (ThermoFisher, Waltham, MA; 88-7503-22) according to manufacturer’s instructions. Example 2 Nanofibers are minimally reactogenic in allergic mice, even when displaying allergenic epitopes

[0078] As a first step to evaluating the self-assembling peptide nanofiber vaccine system as an allergen vaccine platform in vivo reactogenicity of a model nanofiber allergen vaccine was compared to a model protein allergen vaccine (FIG.1)( Cossette BJ, et al. Self- Assembling Allergen Vaccine Platform Raises Therapeutic Allergen-Specific IgG Responses without Induction of Systemic Allergic Responses. ACS Biomater. Sci. Eng.2024, 10, 1819- 1829, incorporated herein by reference). To form the model nanofiber vaccines, a model allergen B cell epitope (OVA61-68) and the self-assembling peptide Q11 were synthesized in tandem (FIGS.4A-4B, TABLE 1). To form a comparative protein-based vaccine against the same model allergen epitope, streptavidin tetramers were loaded with biotinylated OVA61-68peptide (FIG.4C), a strategy used by others to generate multivalent model allergen proteins (Bucaite et al., The Journal of Immunology 2019; 203 (7): 1693-1700). Next, whether either vaccine would elicit an allergic response when used to immunize allergic mice was assessed. Mice were sensitized to OVA61-68via intravenous injection of an anti-OVA61-68IgE mAb (E-C1) and the model allergen vaccines were injected subcutaneously the next day (FIG.4D). It was found that the model protein vaccine induced decreases in body temperature and spikes in serum levels of mast cell protease 1 (MCPT-1), indicating a systemic anaphylactic response. Contrastingly, mice receiving the model nanofiber vaccine showed no signs of a systemic allergic response (FIGS.4E-4F).Docket No.028193-0048-WO01 / 8355

[0079] Next, whether the difference in in vivo reactogenicity could be explained by differences in vaccine allergenicity was assessed. Here, the term “allergenicity” refers to the inherent ability of the vaccine to activate allergen-sensitized mast cells and trigger Type I hypersensitivity responses. It was found that both vaccines were similarly recognized by E- C1 IgE antibody (FIGS.5A-5D) and that both vaccines elicited mast cell activation when exposed to E-C1-sensitized bone marrow-derived mast cells (BMMC) (FIGS.5E-5H). Furthermore, it was found that both vaccines activated peritoneal mast cells when injected directly into the peritoneal cavity of E-C1-sensntized mice (FIGS.5I-5L). Thus, the observed differences in in vivo reactogenicity between the two vaccine types cannot be explained by differential vaccine allergenicity, as both the model nanofiber and protein vaccines were recognized by sIgE and activated allergen-sensitized mast cells.

[0080] Because both model nanofiber and protein allergen vaccines activate mast cells ex vivo and when injected intraperitoneally, but only protein allergen vaccines induce systemic allergic responses after subcutaneous injection, whether this difference could be explained by reduced systemic exposure was assessed. To track systemic vaccine exposure, nanofiber and protein vaccines were labeled with fluorescent dye and fluorescence-matched vaccines were injected subcutaneously into mice. Blood samples were collected over a 24-hour period and it was found that nanofiber vaccines had drastically reduced accumulation in the vascular compartment compared to protein vaccines (FIGS. 6A-6C).

[0081] Local allergic responses can increase vascular permeability and may affect levels of systemic vaccine exposure, so the presence of vaccine material in the bloodstream after injection into E-C1-sensitized mice was tracked (FIG.6D). Protein allergen vaccines induced systemic anaphylactic reactions that were fatal in 60% of mice, whereas nanofiber vaccines did not, despite both vaccines delivering equimolar doses of allergen (FIGS.6E- 6G). Nanofiber vaccines again drastically reduced systemic vaccine exposure compared to protein vaccines, even in the allergic setting (FIGS.6H-6I). Example 3 A model nanofiber allergen vaccine is immunogenic and protects against allergen challenge

[0082] A key feature of a successful allergen vaccine is the ability to generate therapeutic IgG antibody that can neutralize the allergen and prevent Type I hypersensitivity reactions after allergen exposure. Here, the ability of the nanofiber vaccine platform to raiseDocket No.028193-0048-WO01 / 8355 therapeutic sIgG responses was evaluated. (OVA61-68)Q11 nanofiber vaccines alone or together with CpG adjuvant raised high titers of sIgG after only two immunizations (FIGS. 7A-7C). IgG purified from the serum of mice immunized with CpG-adjuvanted (OVA61-68)Q11 vaccines desensitized BMMC to allergen exposure ex vivo, while IgG purified from the serum of naïve mice or mice immunized with epitope-free Q11 nanofibers had no desensitizing effect (FIGS.7D-7E). In this assay, IgG purified from the serum of mice immunized with unadjuvanted (OVA61-68)Q11 reduced BMMC activation but not to a statistically significant level, potentially due to the lower titer of sIgG in this group compared to the CpG-adjuvanted group. Given the allergen-neutralizing ability of sIgG from mice immunized with CpG-adjuvanted (OVA61-68)Q11 vaccines, whether immunized mice would be protected from allergen challenge was assessed. The mice were passively sensitized to OVA61-68and challenged via intraperitoneal injection of model protein allergen (FIG.7F). Naïve mice with no sIgG showed elevated serum levels of MCPT-1 after allergen challenge, whereas mice that received the allergen vaccine did not, suggesting that nanofiber allergen vaccination can protect against Type I allergic responses after allergen exposure (FIG.7G). Example 4 Nanofiber allergen vaccines directed against broadly reactive peanut allergen epitopes are immunogenic and generate therapeutic sIgG

[0083] Motivated by the foregoing findings regarding the safety and efficacy of the model nanofiber allergen vaccines, next allergen epitope targets relevant to human allergic disease were identified and validated. Here, peanut allergy was of focus, one of the most life- threatening food allergies. To identify candidate epitope targets from the major peanut allergen proteins Ara h 1, 2, 3, and 7, six immunodominant epitopes were selected from the literature (TABLE 1) with broad IgE reactivity across allergic patients and their allergenicity was validated using a humanized mast cell line sensitized with serum from peanut allergic human donors (FIGS.8A-8B). Of the six epitopes, four were potently allergenic across all donors (AH1a, AH2a, AH2b, AH3a). The remaining two epitopes (AH1b and AH7a) also triggered detectable activation above baseline in this assay, but not to a statistically significant degree. Next, the immunogenicity of nanofiber vaccines directed against the six candidate epitopes was confirmed, with all vaccines raising robust IgG antibody responses that persisted for at least 25 weeks, which was the latest timepoint that was assayed (FIG. 8C). Additionally, vaccine-induced sIgG recognition of both the peptide antigen as well as the full protein allergen was confirmed (FIGS.8D-8E).Docket No.028193-0048-WO01 / 8355

[0084] To characterize therapeutic potential of sIgG for each target epitope, ex vivo BMMC activation assays were employed (FIG.8F). First, whether vaccine-elicited sIgG could neutralize allergen and desensitize BMMC that were sensitized with serum from mice sensitized against crude peanut extract was assessed. When crude extract sensitized BMMC were exposed to Ara h 1 allergen protein, AH1b sIgG, but not AH1a sIgG, markedly reduced BMMC activation (FIG.8G). When crude extract sensitized BMMC were exposed to Ara h 3 allergen protein, AH3a sIgG also markedly reduced BMMC activation (FIG.8H). For the Ara h 2 and 7 allergens, in this assay the peanut-allergic mice did not raise sufficient IgE responses to sensitize BMMC (FIGS.9A-9C). So, to evaluate sIgG against Ara h 2 and 7, mice were sensitized specifically to these allergens and BMMC with the resulting serum. Here, it was found that IgG from mice immunized against AH7a markedly desensitized BMMC to Ara h 7 exposure (FIG.8I). It was found that serum from mice sensitized to Ara h 2 was unable to sensitize BMMC to the allergen (FIG.9D) and were unable to evaluate the allergen neutralization of AH2a and AH2b sIgG.

[0085] In sum, from the panel of broadly reactive IgE-binding epitopes, it was found that epitopes AH1b, AH3a, and AH7a induced allergen neutralizing sIgG when used as immunogens on nanofiber allergen vaccines. These results confirm that peanut sIgG generated by nanofiber allergen vaccines has therapeutic potential to neutralize allergen and reduce mast cell activation. Interestingly, sIgG from each allergen epitope was tested separately, indicating that sIgG directed against only a single epitope can potently neutralize allergen proteins and prevent mast cell activation even when the cells are sensitized with polyclonal serum. Because the mice did not raise IgE responses to Ara h 2, the therapeutic potential of AH2a or AH2b sIgG could not be evaluated. Example 5 Discussion

[0086] Motivated by the evidence of sIgG involvement in allergen immunotherapy and by promising reports of sIgG mAb therapy efficacy, allergen B cell vaccines aim to elicit long- lived allergen-neutralizing sIgG responses with only a few doses. However, like AIT, these technologies suffer from frequent adverse events caused by patient allergic responses to the vaccine. Here, an initial evaluation of a supramolecular nanofiber vaccine system as an allergen vaccine platform was performed.

[0087] It was found that nanofiber vaccines displaying a model B cell allergen epitope can be administered subcutaneously to allergen sensitized mice without inducing systemicDocket No.028193-0048-WO01 / 8355 allergic responses. In contrast, mice experience systemic anaphylactic reactions if the same allergen epitope is delivered via a protein carrier. Mechanistically, it was found that this behavior is not explained by the inherent allergenicity of the two vaccine types, but rather that nanofiber vaccines prevent leakage of vaccine material into the blood stream. This feature may underpin the reduced systemic reactogenicity of the nanofiber vaccines as systemic allergen exposure is thought to be required for induction of systemic anaphylaxis. Given that particle size is an important determinate of vascular access and lymphatic trafficking, a current hypothesis is that nanofiber vaccine particles are too large to cross the vascular endothelium and are restricted to the interstitial space at the injection site and to lymphatic vessels. Indeed, Q11 nanofiber vaccines remain at the injection site for days. Exclusion from the vascular compartment is likely not unique to Q11 nanofiber vaccines and could affect the safety profile of other nanoparticle-based allergen vaccines – including those based on polymeric particles, liposomes, or VLPs. However, there is at least one example of a clinically tested liposomal allergen vaccine that saw high rates of systemic adverse events, although it is not clear if this was caused by contamination with soluble allergen, allergen adsorption to the liposomal surface, liposomal rupture and allergen release after injection, or by some other mechanism. Further, because the nanofiber vaccine retains allergenicity ex vivo and triggers local mast cell activation after intraperitoneal injection, it is likely that some level of local reactogenicity occurs after subcutaneous injection, although this study did not characterize that. Investigation into local reactions to nanofiber allergen vaccines is warranted and development of strategies to combat local reactions may be of interest.

[0088] The nanofiber vaccine system raises robust and long-lived IgG responses against peptide antigens. Here, it is shown that Q11 nanofiber vaccines directed against six different peptide B cell epitopes from major peanut allergens raise high sIgG titers that persist in mice for at least 25 weeks. For epitope targets from Ara h 1, 3, and 7, it was found that sIgG from immunized mice significantly reduced activation of BMMC. Interestingly, this effect was observed despite BMMC being sensitized with polyclonal allergic sera and the sIgG being directed towards only a single epitope within the allergen protein. As such, the results suggest that an allergen B cell vaccine does not necessarily need to target every IgE-binding epitope on the allergen(s) and that sIgG directed against only a single allergen epitope can achieve desensitization to the entire protein. This effect is likely mediated throughengagement of inhibitory Fc RIIb on BMMC, which can inhibit Fc RI signaling even in trans.However, this effect does also seem to depend on the epitope target as sIgG directed against the AH1a epitope did not neutralize allergen. Interestingly, sIgE in the allergic serum used for this assay recognized the AH1b epitope, but not the AH1a epitope (FIGS.10A-Docket No.028193-0048-WO01 / 8355 10B), suggesting that direct competition for sIgE epitopes is also an important mechanistic component of sIgG allergen neutralization. Unfortunately, mice were unable to be sensitized to the major peanut allergen Ara h 2 and thus the therapeutic potential of vaccines against two of the target epitopes in this study could not be evaluated. However, Ara h 2 is one of the most important allergens in human peanut allergy and these two epitope targets warrant further investigation as potential vaccine targets.

[0089] T cell mediated Type IV hypersensitivity reactions also contribute to allergic disease. The nanofiber vaccines included only allergen B cell epitopes for induction of sIgG, but it may be interesting to include a T cell component in future work. Indeed, AIT efficacy relies not only on sIgG induction, but also on a shift in allergen specific T cell phenotype away from the pro-allergic Th2 phenotype to a Th1 / Treg phenotype. Additionally, it may be worthwhile to consider a hybrid protocol where nanofiber allergen vaccines are administered first to achieve allergen desensitization, followed by AIT. In fact, the presence of pre-existing sIgG can enhance the efficacy of AIT by driving stronger shifts in T cell phenotype. Thus, pre-vaccination before AIT might enhance both the safety and efficacy of a subsequent AIT protocol.

[0090] Overall, the results support further investigation and development of nanofiber allergen vaccines. Allergen delivery via nanofiber vaccines prevents systemic exposure of vaccine material, potentially offering safety advantages over other vaccine platforms, particularly those employing soluble protein allergen. Six epitope targets from peanut allergens were identified that were selected for immunodominance and broad reactivity across peanut allergic individuals and the therapeutic potential of sIgG elicited by nanofiber allergen vaccines directed against three of these targets was confirmed. In future work, development of allergen vaccines will be continued using this platform. Example 6 Immunogen configurations with high inter-epitope spacing are hypoallergenic

[0091] Previous studies of inter-epitope spacing in the context of mast cell activation have been conducted in vitro using the RBL-2H3 cell line, however it is not clear if the same spacing-dependent phenomenon persists in the physiologic setting. To explore the effects of inter-epitope spacing in a physiologically relevant setting, in vitro assays of murine primary bone marrow-derived mast cells (BMMC) and in vivo assays of peritoneal mast cell activation were employed (FIG.11). For the tests, a series of SST model immunogens were designed based on a six-helix-bundle SST (6HB) structure to present the hapten modelDocket No.028193-0048-WO01 / 8355 allergen epitope 2,4-Dinitrophenol (DNP) in one-dimensional configurations that ranged in both valency and inter-epitope spacing (FIGS.12A-12G, FIGS.13A-13C). First, the successful formulation of SST structures with different valencies was verified by attaching and imaging streptavidin molecules (FIGS.12D-12F). Then, four 4xDNP model immunogen structures with different inter-epitope spacings were assembled for subsequent experiments (FIG.12G).

[0092] To elucidate the effects of valency and spacing on mast cell activation, bone marrow-derived mast cells (BMMC) were sensitized with monoclonal mouse anti-DNP IgE (SPE-7) antibody and stimulated with SST model immunogens (FIG.14A). To quantify BMMC activation, the surface expression levels of CD63, a secretory lysosomal protein that reaches the surface of mast cells after activation-induced degranulation, was measured. Virtually no BMMC activation from bivalent (2xDNP) immunogens was observed (FIGS.14B- 14C), but a strong effect of inter-epitope spacing for trivalent (3xDNP) and tetravalent (4xDNP) immunogens was found (FIGS.14D-14G). DNP epitopes separated by 7 nm elicited the highest BMMC activation levels and DNP epitopes separated by 28 nm elicited markedly reduced activation. Increased epitope spacing decreased both the proportion of BMMC that degranulated, as well as the magnitude of degranulation for those that did (FIGS.14H-14I). Additionally, the degree of IgE binding was similar across 4xDNP immunogen configurations and was not correlated to activation (FIGS.15A-15B), suggestingthat inter-epitope spacing impacts signaling downstream of Fc RI.

[0093] Factors present in vivo can alter mast cell sensitivity to allergen. To investigate if mast cell sensitivity to inter-epitope spacing persists in vivo (in a model that minimizes requirements for extended durability of the immunogens) mice were passively sensitized to DNP by injecting SPE-7 IgE and then they were intraperitoneally challenged with 4xDNP SST model immunogens (FIG.14J). In line with Applicants’ previous results, peritoneal mast cells were strongly activated by 4x7nm SST model immunogens, and 4x28nm immunogens elicited markedly lower levels of activation (FIGS.14K-14L). Together, these results suggest that multivalent immunogens bearing highly IgE-reactive epitopes can be made hypoallergenic by increasing inter-epitope spacing. TABLE 5 shows P values for FIGS.14B-14L.Docket No.028193-0048-WO01 / 8355Docket No.028193-0048-WO01 / 8355 Example 7 B cells are activated by hypoallergenic immunogen configurations

[0094] A recent study of B cell activation found that unlike in mast cells, stronger signaling was triggered as antigens were spaced further apart. However, the VRC01 / eOD BCR / antigen system employed in the previous study was considerably different than the allergen / IgE systems used in other previous mast cell studies, including Applicants’ described above. Because differences in antibody affinity and structure can influence B cell signaling, whether a similar trend in B cells using a BCR / antigen system that is analogous to the DNP / SPE-7 allergen / IgE system employed in the mast cell experiments would be observed. For this, a chimeric anti-DNP IgM BCR was designed by fusing the variable regions of the mouse SPE-7 IgE antibody to the constant regions of human membrane- bound IgM and generated a line of RAMOS B cells that stably express the transgenic BCR (RAMOSDNP) (FIGS.16A-16D).

[0095] B cell activation through the BCR is characterized by an early and rapid increase in cytosolic calcium and subsequent downstream activation of multiple signaling pathways, including the RAS-ERK pathway. To assess the effect of inter-epitope spacing on downstream B cell signaling, ERK phosphorylation (pERK) in RAMOSDNP cells was measured after stimulation with 4xDNP SST model immunogens (FIG.17A). Regardless of inter-epitope spacing, 4xDNP SST immunogens triggered similar levels of pERK after 5 minutes of stimulation (FIG.17B). Additionally, pERK levels were similar in magnitude to those achieved with DNP-BSA stimulation and were uniform across the entire B cell population (FIG.17C). When compared to normalized mast cell activation, normalized B cell activation was ~10-fold higher in response to 4x28nm SST model immunogens (FIG.17D).

[0096] While RAMOSDNP cell activation converged to a maximum after 5 minutes of stimulation, 4xDNP immunogens with high inter-epitope spacing triggered significantly less pERK after only one minute (FIG.17B). To determine how very early signaling events were affected by inter-epitope spacing for immunogens with varying multivalency, calcium flux in RAMOSDNP cells was monitored (FIG.17E) after stimulation with 2xDNP (FIG.17F), 3xDNP (FIG.17G), and 4xDNP (FIG.17H) SST model immunogens. RAMOSDNP cells showed no appreciable early calcium influx for 2xDNP SST immunogens and modest RAMOSDNP activation was triggered in response to 3xDNP SST immunogens, consistent with the dependence of BCR signaling on antigen multivalency. 4xDNP SST model immunogens with 7 nm and 14 nm inter-epitope spacing elicited similar RAMOSDNP cell activation compared to DNA-BSA, but calcium flux was comparatively lower when DNPDocket No.028193-0048-WO01 / 8355 epitopes were separated by 21 nm and 28 nm (FIG.17H). This decrease in total calcium flux signal was due to a lower proportion of activated cells during a given time period (FIGS. 17I-17J), with 4x28nm immunogens triggering elevated calcium levels in ~60% of cells during the stimulation peak. However, within this activated population, calcium signal magnitude was minimally dependent on inter-epitope spacing, and all cells showed calcium levels equivalent or similar to the DNP-BSA positive control (FIG.17K). Given that B cell activation converges as evidenced by pERK levels after 5 minutes of stimulation, it is possible that immunogens with more distant inter-epitope spacing take more time to accumulate on the B cell surface, but ultimately achieve sufficient binding to trigger robust B cell activation. TABLE 6 shows P values for FIGS.17B-17K.Docket No.028193-0048-WO01 / 8355Docket No.028193-0048-WO01 / 8355Docket No.028193-0048-WO01 / 8355

[0097] Taken together, these results suggest that early B cell activation kinetics are slowed when epitopes are separated by large distances, but that B cells ultimately achieve near-maximal activation regardless of inter-epitope spacing. Overall, by using a single unified system to assess the effects of inter-epitope spacing on both mast cell and B cell activation, the results support the core principle of the immunogen design strategy herein: that epitope spacing distances can be optimized for B cell activation in the absence of mast cell activation. Example 8 Supramolecular peptide nanofibers enable tunable control over epitope density via modulation of peptide subunit stoichiometry

[0098] Encouraged by the foregoing results, hypoallergenic allergen vaccine immunogens were designed for testing in models of allergic disease. Because unmodified DNA-based materials suffer from poor in vivo stability and are relatively unestablished for use as immunogens, supramolecular nanofiber immunogens well established by Applicants as potent self-adjuvating particles that raise robust antibody responses against peptide epitopes presented on the fiber surface were used. The -helical self-assembling peptideDocket No.028193-0048-WO01 / 8355 Coil29 (C29; H2N-QARILEADAEILRAYARILEAHAEILRAQ-Am (SEQ ID NO: 9)) forms nanofibers in which the helical peptides run perpendicular to the fiber axis. Peptide epitopes can be synthesized in tandem to the N-termini of the C29 subunits and then assembled into nanofiber vaccine immunogens. The density of peptide epitopes can be controlled by varying the stoichiometric ratio of peptide-C29 to naked C29 subunits, and that by doing so, the average distance between epitopes along the nanofiber can be manipulated (FIG.18A). Of critical importance is the capacity for long and narrow nanomaterials such as C29 nanofibers to establish significant distances between epitopes. In the case of C29 nanofibers, this was achieved by diluting one peptide (the epitope bearing C29) into an unmodified background to distance the epitopes from each other along the long axis of the nanofiber. This capability would be challenging in spherical nanoparticles, carrier proteins, or other shapes lacking a long axis.

[0099] First, whether peptide epitopes could be reliably co-assembled within C29 immunogens was tested. a model peptide epitope from ovalbumin (OVA323-329) was synthesized in tandem with C29 and labeled the N-terminus with either FITC or TAMRA fluorophores. Then, TAMRA-only, FITC-only, or co-assembled nanofibers with each fluorophore-labeled subunit species at a molar ratio of 10% in a background of unmodified C29 peptide were assembled, and the FITC, TAMRA, or FRET signal of individual nanofibers was measured using confocal microscopy (FIGS.18B-18D). It was reasoned that uniform co-assembly of the two labeled peptides would produce detectable FRET signatures for (TAMRA-OVA10FITC-OVA10)C29 nanofibers but not for single-fluorophore nanofibers (TAMRA-OVA)10C29 or (FITC-OVA)10C29 (FIGS.18E-18G) (subscript denotes percentage of peptide species). Consistent with this hypothesis, a strong FITC signal was observed from (FITC-OVA)10C29 nanofibers that was significantly dampened in the (TAMRA-OVA10FITC-OVA10)C29 formulation (FIG.18E), indicative of FRET quenching and co-assembly of the two labeled peptides. Also, a FRET signal was observed in nanofibers that was absent in (TAMRA-OVA)10C29 and (FITC-OVA)10C29 nanofiber formulations, further supporting the ability to produce nanofiber immunogens bearing multiple co- assembled peptide epitopes (FIG.18G). [000100] Next, whether the epitope density on nanofiber immunogens could be controlled by changing the stoichiometric ratio of epitope-C29 to C29 was assessed. C29 nanofibers were formulated with 10%, 2.5%, or 1.25% (TAMRA-OVA)C29 and the TAMRA signal was quantified for individual nanofibers, again using confocal microscopy. As expected, titration of (TAMRA-OVA)C29 into nanofibers resulted in distinct distributions of TAMRA intensity, each with an average brightness that decreased corresponding to (TAMRA-OVA)C29Docket No.028193-0048-WO01 / 8355 stoichiometry (FIG.19, FIG.18H). Distributions were polydisperse, indicating less precise control over epitope density using this approach compared with DNA SST, but distributions became tighter as the labeled peptide was diluted into the nanofibers (FIG.18H). Together, these results demonstrate the ability to tune peptide epitope density - and in turn, control the average inter-epitope spacing – of supramolecular nanofiber immunogens via epitope co- assembly. TABLE 7 shows P values for FIGS.18I-18K.Example 9 Coil29 nanofiber immunogens with ultra-low epitope density are hypoallergenic and trigger B cell activation [000101] Having established that functionalized C29 peptides could be co-assembled and diluted extensively within supramolecular peptide immunogens, whether ultra-low epitope density, and consequently high inter-epitope spacing, could avoid mast cell activation while maintaining B cell reactivity was explored. For this investigation, BMMCs, RAMOSDNP cells, and the same activation assays as described above for SST immunogens were employed. First, a series of nanofiber immunogens with decreasing epitope density was formulated by varying the ratio of (DNP)C29:C29 from 25% (1:4 dilution of the epitope into the C29 nanofibers) down to 0.2% (1:500 dilution of the epitope into the C29 nanofibers). The distance between epitopes on nanofibers in this range are estimated to range from 0.88 nm (at 25% epitope) to 110 nm (at 0.2% epitope) (FIGS.20A-20B). Then, the reactivity ofDocket No.028193-0048-WO01 / 8355 these nanofibers towards DNP-sensitized BMMC was analyzed (FIG.21A). (DNP)1C29 nanofibers (1% DNP, 1:100 dilution of epitope) showed markedly reduced allergenicity, and (DNP)0.5C29 and (DNP)0.2C29 nanofibers did not trigger any detectable BMMC activation at all (FIGS.21B-21C). To establish that this strategy applies beyond the hapten DNP, BMMC were sensitized with mouse anti-OVA61-68IgE (E-C1) and stimulated with nanofibers containing the peptide allergen epitope OVA61-68rather than DNP. Similar to previous results, ultra-low density (OVA61-68)C29 nanofibers avoided any detectable BMMC activation (FIGS.21D-21E). [000102] In both allergen systems, nanofibers formulated with 0.5% epitope-C29 peptide completely avoided mast cell activation, so the extent to which (DNP)0.5C29 could activate B cells (RAMOSDNP) was investigated. Nanofibers formulated with epitope contents of both 5% (1:20 dilution of epitope) and 0.5% (1:200 dilution) triggered strong B cell activation in at least 85% of RAMOSDNP B cells at equimolar epitope concentrations (FIGS.21F-21I), although the magnitude of pERK expression was slightly lower in cells stimulated with the low-density formulation (FIG.21I). Interestingly, these results aligned closely with Applicants’ previous findings using DNA SST model immunogens, in that an average inter- epitope spacing of 28 nm along the peptide fiber axis is expected at about 0.8% epitope content (FIGS.20A-20B). This spacing is consistent with those identified using DNA SST that activated B cells but not mast cells. TABLE 8 shows P values for FIGS.21B and 21D.Docket No.028193-0048-WO01 / 8355Docket No.028193-0048-WO01 / 8355 0.2% vs.0%Example 10 Vaccination with hypoallergenic nanofiber immunogens elicits allergen-specific IgG responses that protect against allergic responses [000103] Next, hypoallergenic nanofiber allergen vaccine performance was evaluated in vivo by asking if they could raise allergen-neutralizing IgG responses that protect against a subsequent allergen challenge. Therapeutic allergen vaccination is performed in allergic patients that have pre-existing anti-allergen immunity, which includes allergen specific IgG+ memory B cells. To simulate allergic immunological memory, an adoptive transfer model was employed. First, mice were actively sensitized to the OVA61-68allergen epitope via repeated intraperitoneal injections with a KLH conjugate and AlHydrogel (FIG.22A). Three weeks later, splenocytes from OVA61-68-sensitized mice were adoptively transferred into naïve recipient mice. The following day, mice were immunized with (OVA61-68)C29 immunogens (FIG.23) with different epitope densities and monitored for allergen specific IgG. Regardless of epitope density, all (OVA61-68)C29 immunogens triggered allergen specific antibody responses, although the high density (OVA61-68)25C29 elicited significantly higher responses (FIGS.22B-22C). Because the immunostimulatory adjuvant effect of nanofiber immunogens is inherent, equal doses of nanofiber was delivered, rather than matching the epitope dose across formulations. Thus, low density formulations contained between 25- to 100-fold less epitope per immunization. It is possible that the immunogenicity of low-density formulations is impacted by the reduced epitope dose. Further, nanofiber valency and epitope spacing cannot be manipulated independent of one another, and the high-density formulation consequently has higher epitope valency, which also may contribute the enhanced immunogenicity. Additionally, for protein nanoparticle immunogens, it has been shown that if scaffold is provided in excess, antigen-specific antibody responses can be competitively inhibited. Indeed, compared to (OVA61-68)25C29, ultra-low density formulations elicited greater anti-C29 IgG antibody responses (FIG.24), and this phenomenon may play a role as well. [000104] Despite differences in antibody response magnitude, IgG purified from the serum of all immunization groups potently neutralized model allergen and prevented allergen induced BMMC activation in vitro (FIG.22D). To test if the allergen specific antibody responses could protect against allergen challenge in vivo, the mice were passively sensitized against OVA61-68via intravenous injection of E-C1 IgE and challenged them theDocket No.028193-0048-WO01 / 8355 following day with a model allergen OVA61-68tetramer (FIG.22A). Compared to naïve mice, mice immunized with both (OVA61-68)25C29 and (OVA61-68)0.5C29 showed reduced drops in body temperature (FIG.22E-22F) and lower serum concentrations of mast cell protease 1 (MCPT-1) (FIG.22G-22J) after challenge. Although, while mice immunized with (OVA61-68)25C29 were completely protected, immunization with (OVA61-68)0.5C29 achieved only partial protection from anaphylaxis, likely due to the lower antibody responses elicited by these immunogens. Together these results provide proof-of-concept for the approach herein to hypoallergenic immunogen design and highlight key areas for the future improvement of this strategy. TABLE 9 shows P values for FIGS.22C-22J.Docket No.028193-0048-WO01 / 8355Example 11 Discussion II [000105] This study provides proof-of-concept for a novel mode of hypoallergenic immunogen design based on nanoscale control of the space between IgE-reactive allergen epitopes. While many modes of hypoallergenic immunogen design focus on the obviation of IgE-reactive epitopes, this strategy allows for immunization specifically against these epitopes, which may improve allergen neutralizing antibody responses while limiting allergic reactogenicity when used for therapeutic allergen vaccination. [000106] While the in vitro results of mast cell and B cell activation using model DNA SST- based immunogens were largely consistent with previous studies, some key differences in the findings herein were observed. In the case of mast cells, little to no activation from bivalent ligands was observed, in contrast to previous reports of mast cell activation against bivalent ligands using the RBL-2H3 cell line. However, RBL-2H3 activation in response to bivalent ligands was low compared to trivalent ligands, which is consistent with the findings herein. Indeed, in the above mentioned study, an inhibitor of actin polymerization (cytochalasin D) was used to achieve meaningful mast cell activation in response to bivalentligands by preventing Fc RI internalization to enhance sensitivity. In other contexts, bivalentDocket No.028193-0048-WO01 / 8355 ligands have been shown to be poor activators of mast cells due to the potential for formation of cyclic IgE-allergen complexes and have even been explored as inhibitors of mast cell activation. It is also important to reiterate that while the studies herein employed the same SPE-7 / DNP IgE / allergen system used in these previous reports, primary BMMC was utilized herein, which may also contribute to differences in the effects of bivalent ligands. In the case of B cells, two main discrepancies from similar studies previously performed with RAMOS cells were observed. In contrast to previous reports, no detectable activation of RAMOSDNP cells in response to bivalent model immunogens was observed. Additionally, while previous reports described increasing B cell activation strength with increasing inter-epitope spacing, herein, activation kinetics were slowed as spacing increased and B cells ultimately converged to near-maximum activation levels regardless of inter-epitope spacing. One possible source of these discrepancies is that the VRC01 / eOD BCR / antigen system used in these studies represents a much higher affinity interaction (KD 30 pM) than the SPE-7 / DNP BCR / antigen system employed herein (KD 20 nM), which may alter B cell sensitivity to low-valency antigen. [000107] Reductionist models of IgE-related allergy that rely on a single monoclonal interaction between the allergen and IgE were employed. While these models provide excellent sensitivity and control over experimental parameters, some features of clinical allergic disease are not recapitulated in this setting, including the presence of polyclonal anti- allergen IgE responses. Considerable effort may be required to identify allergen B cell epitope targets that are compatible with the immunogen design approach herein. For example, longer target peptides containing multiple epitopes that could be occupied by multiple IgE antibodies simultaneously may need to be excluded. Additionally, unlike true allergic disease, the models herein focused on Type I hypersensitivity and thus lacked allergen-specific T cells, which can drive allergic symptoms via Type IV hypersensitivity reactions. The vaccines herein did not include allergen-derived T cell epitopes and were designed only to raise therapeutic IgG responses. It is not immediately clear if these responses would impact desensitization against Type IV hypersensitivity reactions, however other allergen vaccines have employed a similar “B cell only” strategy with encouraging results. In further support of this strategy, studies of therapeutic allergen-specific monoclonal IgG antibodies suggest that the presence of allergen neutralizing IgG alone can achieve meaningful allergic desensitization. [000108] To study mast cell responses, models of in vitro and in vivo primary mast cell activation were employed. However, the B cell studies herein were limited to assays using the RAMOSDNP model, which is based on an immortalized cell line. It cannot be ruled outDocket No.028193-0048-WO01 / 8355 that primary B cells exhibit different sensitivity to inter-epitope spacing compared to RAMOSDNP B cells, although hypoallergenic nanofiber immunogens triggered B cell- mediated antibody responses in vivo. Additionally, germline B cells may respond differently than memory B cells either due to differences in BCR affinity or effects of disparate differentiation states. Assessing the impact of inter-epitope spacing on primary germline and memory B cells could be an interesting area of future study. [000109] Finally, while the nanofiber immunogens utilized in this study allow for coarse control over epitope density, they do possess limitations that may limit the efficacy of the hypoallergenic immunogen design approach herein. For example, epitope density and valency cannot be controlled independently, and epitope spacing on the fiber can only be controlled on average. Additionally, the ultra-low density nanofiber vaccines herein generated therapeutic allergen neutralizing antibody responses but were less immunogenic compared to high-density formulations. Ultra-low density nanofibers (0.5% epitope) contain 100-fold less epitope compared to the formulations normally used by Applicants (50% epitope), and this was not controlled for in the immunogenicity study herein because it was more prudent to dose-match the total nanofiber content, rather than the epitope content. Because peptide nanofibers are large particles and can remain at the injection site for days when dosed subcutaneously, it is likely that vaccine material is slow to accumulate in the draining lymph nodes, which may limit antigen availability and compromise immunogenicity for low-epitope content nanofibers. To combat this issue, it may be wise to use a vaccine scaffold system with a particulate size of ~100-200 nm diameter, which readily and rapidly drain through lymphatic vessels to draining lymph nodes. A platform combining optimized immunogen size and independent control of epitope spacing and valency, may be particularly advantageous for this mode of hypoallergenic immunogen design. [000110] It was hypothesized that IgE-reactive immunogens could be made hypoallergenic using a nanoscale immunogen design strategy to separate allergen epitopes at optimized distances. Using DNA-based tools with precise control over nanoscale spatial parameters, the core principle of this hypothesis was validated by showing that mast cell activation was attenuated when the epitopes of multivalent immunogens were separated by 28 nm while B cells remained responsive to the same immunogen configuration. Based on these findings, hypoallergenic supramolecular peptide nanofiber immunogens that displayed IgE-reactive allergen epitopes without triggering mast cell activation were designed. When used for allergen vaccination, these immunogens generated allergen-specific IgG responses that neutralized allergen and diminished hypersensitivity responses to allergen challenge.Docket No.028193-0048-WO01 / 8355 Together these results provide proof-of-concept for a novel strategy of hypoallergenic immunogen design with unique capabilities. *** [000127] The foregoing description of the specific aspects will so fully reveal the general nature of the invention that others can, by applying knowledge within the skill of the art, readily modify and / or adapt for various applications such specific aspects, without undue experimentation, without departing from the general concept of the present disclosure. Therefore, such adaptations and modifications are intended to be within the meaning and range of equivalents of the disclosed aspects, based on the teaching and guidance presented herein. It is to be understood that the phraseology or terminology herein is for the purpose of description and not of limitation, such that the terminology or phraseology of the present specification is to be interpreted by the skilled artisan in light of the teachings and guidance. [000128] The breadth and scope of the present disclosure should not be limited by any of the above-described exemplary aspects, but should be defined only in accordance with the following claims and their equivalents. [000129] All publications, patents, patent applications, and / or other documents cited in this application are incorporated by reference in their entirety for all purposes to the same extent as if each individual publication, patent, patent application, and / or other document were individually indicated to be incorporated by reference for all purposes. [000130] For reasons of completeness, various aspects of the invention are set out in the following numbered clauses: [000131] Clause 1. An allergen conjugate peptide comprising: (i) a self-assembling peptide comprising a polypeptide having the amino acid sequence of bXXXb (SEQ ID NO: 1, wherein X is independently any amino acid and b is independently any positively charged amino acid), QQKFQFQFEQQ (SEQ ID NO: 12), FKFEFKFE (SEQ ID NO: 14), KFQFQFE (SEQ ID NO: 15), QQRFQFQFEQQ (SEQ ID NO: 16), QQRFQWQFEQQ (SEQ ID NO: 17), FEFEFKFKFEFEFKFK (SEQ ID NO: 18), QQRFEWEFEQQ (SEQ ID NO: 19), QQXFXWXFQQQ (SEQ ID NO: 20, where X is ornithine), FKFEFKFEFKFE (SEQ ID NO: 21), FKFQFKFQFKFQ (SEQ ID NO: 22), AEAKAEAKAEAKAEAK (SEQ ID NO: 23), AEAEAKAKAEAEAKAK (SEQ ID NO: 24), AEAEAEAEAKAKAKAK (SEQ ID NO: 25), RADARADARADARADA (SEQ ID NO: 26), RARADADARARADADA (SEQ ID NO: 27), SGRGYBLGGQGAGAAAAAGGAGQGGYGGLGSQG (SEQ ID NO: 28), EWEXEXEXEXDocket No.028193-0048-WO01 / 8355 (SEQ ID NO: 29, where X is Val, Ala, Ser, or Pro), WKXKXKXKXK (SEQ ID NO: 30, where X is Val, Ala, Ser, or Pro), KWKVKVKVKVKVKVK (SEQ ID NO: 31, where X is Val, A, Ser, or Pro), LLLLKKKKKKKKLLLL (SEQ ID NO: 32), VKVKVKVKVDPPTKVKVKVKV (SEQ ID NO: 33), VKVKVKVKVDPPTKVKTKVKV (SEQ ID NO: 34), KVKVKVKVKDPPSVKVKVKVK (SEQ ID NO: 35), VKVKVKVKVDPPSKVKVKVKV (SEQ ID NO: 36), VKVKVKTKVDPPTKVKTKVKV (SEQ ID NO: 37), QQKFxFQFEQQ (SEQ ID NO: 38, wherein x is Glu, Asp, or Asn), QQKFQxQFEQQ (SEQ ID NO: 39, wherein x is Trp or Tyr), or QQKFQFxFEQQ (SEQ ID NO: 40, wherein x is Glu, Asp, or Asn); and (ii) at least one allergen epitope conjugated to a terminus of the self-assembling peptide. [000132] Clause 2. The allergen conjugate peptide of clause 1, wherein the allergen epitope is a B cell allergen epitope. [000133] Clause 3. The allergen conjugate peptide of clause 1 or 2, wherein the allergen epitope is IgE-reactive. [000134] Clause 4. The allergen conjugate peptide of any one of clauses 1-3, wherein the allergen epitope comprises a peanut allergen epitope. [000135] Clause 5. The allergen conjugate peptide of clause 4, wherein the peanut allergen epitope is from peanut allergen protein Ara h 1, Ara h 3, or Ara h 7, or Ara h 2. [000136] Clause 6. The allergen conjugate peptide of clause 5, wherein the peanut allergen epitope comprises AH1b, AH3a, AH7a, AH1a, AH2a, or AH2b, or a combination thereof. [000137] Clause 7. The allergen conjugate peptide of clause 6, wherein the peanut allergen epitope comprises AH1b, AH3a, or AH7a, or a combination thereof. [000138] Clause 8. The allergen conjugate peptide of any one of clauses 1-7, wherein each self-assembling peptide forms a beta sheet and comprises a polypeptide having an amino acid sequence selected from QQKFQFQFEQQ (SEQ ID NO: 12), FKFEFKFE (SEQ ID NO: 14), KFQFQFE (SEQ ID NO: 15), QQRFQFQFEQQ (SEQ ID NO: 16), QQRFQWQFEQQ (SEQ ID NO: 17), FEFEFKFKFEFEFKFK (SEQ ID NO: 18), QQRFEWEFEQQ (SEQ ID NO: 19), QQXFXWXFQQQ (SEQ ID NO: 20, where X is ornithine), FKFEFKFEFKFE (SEQ ID NO: 21), FKFQFKFQFKFQ (SEQ ID NO: 22), AEAKAEAKAEAKAEAK (SEQ ID NO: 23), AEAEAKAKAEAEAKAK (SEQ ID NO: 24), AEAEAEAEAKAKAKAK (SEQ ID NO: 25), RADARADARADARADA (SEQ ID NO: 26), RARADADARARADADA (SEQ ID NO: 27),Docket No.028193-0048-WO01 / 8355 SGRGYBLGGQGAGAAAAAGGAGQGGYGGLGSQG (SEQ ID NO: 28), EWEXEXEXEX (SEQ ID NO: 29, where X is Val, Ala, Ser, or Pro), WKXKXKXKXK (SEQ ID NO: 30, where X is Val, Ala, Ser, or Pro), KWKVKVKVKVKVKVK (SEQ ID NO: 31, where X is Val, A, Ser, or Pro), LLLLKKKKKKKKLLLL (SEQ ID NO: 32), VKVKVKVKVDPPTKVKVKVKV (SEQ ID NO: 33), VKVKVKVKVDPPTKVKTKVKV (SEQ ID NO: 34), KVKVKVKVKDPPSVKVKVKVK (SEQ ID NO: 35), VKVKVKVKVDPPSKVKVKVKV (SEQ ID NO: 36), VKVKVKTKVDPPTKVKTKVKV (SEQ ID NO: 37), QQKFxFQFEQQ (SEQ ID NO: 38, wherein x is Glu, Asp, or Asn), QQKFQxQFEQQ (SEQ ID NO: 39, wherein x is Trp or Tyr), and QQKFQFxFEQQ (SEQ ID NO: 40, wherein x is Glu, Asp, or Asn). [000139] Clause 9. The allergen conjugate peptide of any one of clauses 1-7, wherein each self-assembling peptide forms an alpha-helix and comprises a polypeptide having an amino acid sequence of bXXXb (SEQ ID NO: 1), wherein X is independently any amino acid and b is independently any positively charged amino acid. [000140] Clause 10. The allergen conjugate peptide according to clause 8 or 9, wherein the self-assembling peptide comprises the sequence QQKFQFQFEQQ (SEQ ID NO: 12) orAc-QQKFQFQFEQQ-NH2(SEQ ID NO: 13) or bXXXb (SEQ ID NO: 1, wherein X isindependently any amino acid and b is independently any positively charged amino acid). [000141] Clause 11. The allergen conjugate peptide of any one of clauses 1-7 and 9-10, wherein b is independently selected from Arg and Lys. [000142] Clause 12. The allergen conjugate peptide of any one of clauses 1-7 and 9-11, wherein bXXXb (SEQ ID NO: 1) is RAYAR (SEQ ID NO: 2) or KAYAK (SEQ ID NO: 3). [000143] Clause 13. The allergen conjugate peptide of any one of clauses 1-7 and 9-12,wherein the self-assembling peptide comprises an amino acid sequence of ZnbXXXbZm(SEQ ID NO: 5), wherein b is independently any positively charged amino acid, Z is independently any amino acid, X is independently any amino acid, n is an integer from 0 to 20, and m is an integer from 0 to 20. [000144] Clause 14. The allergen conjugate peptide of clause 13, wherein the self- assembling peptide comprises an amino acid sequence selected from QARILEADAEILRAYARILEAHAEILRAQ (Coil29, SEQ ID NO: 6), or QAKILEADAEILKAYAKILEAHAEILKAQ (SEQ ID NO: 7), or ADAEILRAYARILEAHAEILRAQ(SEQ ID NO: 8), or Ac-QARILEADAEILRAYARILEAHAEILRAQ-NH2(SEQ ID NO: 9), or Ac-QAKILEADAEILKAYAKILEAHAEILKAQ-NH2(SEQ ID NO: 10), or Ac-Docket No.028193-0048-WO01 / 8355ADAEILRAYARILEAHAEILRAQ-NH2(SEQ ID NO: 11), or QAEILRAYARILEAQ (SEQ IDNO: 101), or QAEILRAYARILEAHAEILKAQ (SEQ ID NO: 102), or QAEILRAYARILEADAKILEAHAEILKAQ (SEQ ID NO: 103), or QAEILRAYARILEADAEILKAQAKILEAHAEILKAQ (SEQ ID NO: 104). [000145] Clause 15. The allergen conjugate peptide of any one of clauses 1-14, wherein the self-assembling peptide forms a beta sheet and the at least one allergen epitope is attached to the C-terminus or the N-terminus of the self-assembling peptide or a combination thereof, or wherein the self-assembling peptide forms an alpha-helix and the at least one allergen epitope is attached to the N-terminus of the self-assembling peptide. [000146] Clause 16. The allergen conjugate peptide of any one of clauses 1-15, wherein 1 to 10 allergen epitopes are attached to the C-terminus or the N-terminus of the self- assembling peptide. [000147] Clause 17. The allergen conjugate peptide of any one of clauses 1-8 and 15-16, wherein the self-assembling peptide forms a beta sheet and the allergen conjugate peptide further comprises: (iii) a PEG molecule or a PAS peptide conjugated to the self-assembling peptide. [000148] Clause 18. The allergen conjugate peptide of clause 17, wherein the PAS peptide comprises a sequence of Pro-Ala-Ser or comprises the sequence of ASPAAPAPASPAAPAPSAPA (SEQ ID NO: 97), or a peptide having at least 80%, 85%, 90%, or 95% identity thereto. [000149] Clause 19. The allergen conjugate peptide of clause 17, wherein the PEG molecule comprises PEG-2000. [000150] Clause 20. The allergen conjugate of any one of clauses 17-19, wherein the PEG molecule or the PAS peptide is conjugated to the self-assembling peptide at the same or the opposite terminus from wherein the allergen epitope is attached. [000151] Clause 21. The allergen conjugate peptide of any one of clauses 1-20, further comprising: (iv) at least one linker. [000152] Clause 22. The allergen conjugate peptide of clause 21, wherein the at least one linker is between the at least one allergen epitope and the self-assembling peptide. [000153] Clause 23. The allergen conjugate peptide of clause 21, wherein the at least one linker comprises a first linker between the at least one allergen epitope and the self-Docket No.028193-0048-WO01 / 8355 assembling peptide, and a second linker between the PEG molecule or the PAS peptide and the self-assembling peptide. [000154] Clause 24. The allergen conjugate peptide of any one of clauses 21-23, whereinthe at least one linker comprises SEQ ID NO: 83 (SGSG), SEQ ID NO: 84 ((Ser-Gly)2), SEQID NO: 85 (CCCCSGSG), SEQ ID NO: 86 (Gnwherein n is an integer from 1 to 10), SEQ IDNO: 87 (GSGS), SEQ ID NO: 88 (SSSS), SEQ ID NO: 89 (GGGS), SEQ ID NO: 90 (GGC),SEQ ID NO: 91 ((GGC)8), SEQ ID NO: 92 ((G4S)3), SEQ ID NO: 93 (KSGSG), SEQ ID NO:94 (KKSGSG), SEQ ID NO: 95 (EAAAK)2, or SEQ ID NO: 96 (GGAAY). [000155] Clause 25. A nanofiber comprising a plurality of the allergen conjugate peptide of any one of clauses 1-24, wherein the plurality of allergen conjugate peptides self-assemble into the nanofiber. [000156] Clause 26. A nanofiber comprising: (i) at least one allergen conjugate peptide of any one of clauses 1-24; and at least one of a peptide selected from a (ii) T-cell epitope- conjugate peptide, a (iii-a) PEG conjugate peptide, a (iii-b) PAS conjugate peptide, and a (iv) plain self-assembling peptide, or a combination thereof, (ii) wherein the T-cell epitope- conjugate peptide comprises: a self-assembling peptide comprising a polypeptide having the amino acid sequence of QQKFQFQFEQQ (SEQ ID NO: 12), bXXXb (SEQ ID NO: 1, wherein X is independently any amino acid and b is independently any positively charged amino acid), FKFEFKFE (SEQ ID NO: 14), KFQFQFE (SEQ ID NO: 15), QQRFQFQFEQQ (SEQ ID NO: 16), QQRFQWQFEQQ (SEQ ID NO: 17), FEFEFKFKFEFEFKFK (SEQ ID NO: 18), QQRFEWEFEQQ (SEQ ID NO: 19), QQXFXWXFQQQ (SEQ ID NO: 20, where X is ornithine), FKFEFKFEFKFE (SEQ ID NO: 21), FKFQFKFQFKFQ (SEQ ID NO: 22), AEAKAEAKAEAKAEAK (SEQ ID NO: 23), AEAEAKAKAEAEAKAK (SEQ ID NO: 24), AEAEAEAEAKAKAKAK (SEQ ID NO: 25), RADARADARADARADA (SEQ ID NO: 26), RARADADARARADADA (SEQ ID NO: 27), SGRGYBLGGQGAGAAAAAGGAGQGGYGGLGSQG (SEQ ID NO: 28), EWEXEXEXEX (SEQ ID NO: 29, where X is Val, Ala, Ser, or Pro), WKXKXKXKXK (SEQ ID NO: 30, where X is Val, Ala, Ser, or Pro), KWKVKVKVKVKVKVK (SEQ ID NO: 31, where X is Val, A, Ser, or Pro), LLLLKKKKKKKKLLLL (SEQ ID NO: 32), VKVKVKVKVDPPTKVKVKVKV (SEQ ID NO: 33), VKVKVKVKVDPPTKVKTKVKV (SEQ ID NO: 34), KVKVKVKVKDPPSVKVKVKVK (SEQ ID NO: 35), VKVKVKVKVDPPSKVKVKVKV (SEQ ID NO: 36), VKVKVKTKVDPPTKVKTKVKV (SEQ ID NO: 37), QQKFxFQFEQQ (SEQ ID NO: 38, wherein x is Glu, Asp, or Asn), QQKFQxQFEQQ (SEQ ID NO: 39, wherein x is Trp or Tyr), and QQKFQFxFEQQ (SEQ ID NO: 40, wherein x is Glu, Asp, or Asn); and at least one T-Docket No.028193-0048-WO01 / 8355 cell epitope conjugated to a terminus of the self-assembling peptide, wherein the at least one T-cell epitope is selected from PADRE and VAC, and wherein PADRE comprises a polypeptide having the amino acid sequence of aKXVAAWTLKAa (SEQ ID NO: 99, wherein “X” comprises cyclohexylalanine and “a” comprises D-alanine), and wherein VAC comprises a polypeptide having the amino acid sequence of QLVFNSISARALKAY (SEQ ID NO: 100), (iii-a) wherein the PEG conjugate peptide comprises: a self-assembling peptide comprising a polypeptide having the amino acid sequence of QQKFQFQFEQQ (SEQ ID NO: 12), bXXXb (SEQ ID NO: 1, wherein X is independently any amino acid and b is independently any positively charged amino acid), FKFEFKFE (SEQ ID NO: 14), KFQFQFE (SEQ ID NO: 15), QQRFQFQFEQQ (SEQ ID NO: 16), QQRFQWQFEQQ (SEQ ID NO: 17), FEFEFKFKFEFEFKFK (SEQ ID NO: 18), QQRFEWEFEQQ (SEQ ID NO: 19), QQXFXWXFQQQ (SEQ ID NO: 20, where X is ornithine), FKFEFKFEFKFE (SEQ ID NO: 21), FKFQFKFQFKFQ (SEQ ID NO: 22), AEAKAEAKAEAKAEAK (SEQ ID NO: 23), AEAEAKAKAEAEAKAK (SEQ ID NO: 24), AEAEAEAEAKAKAKAK (SEQ ID NO: 25), RADARADARADARADA (SEQ ID NO: 26), RARADADARARADADA (SEQ ID NO: 27), SGRGYBLGGQGAGAAAAAGGAGQGGYGGLGSQG (SEQ ID NO: 28), EWEXEXEXEX (SEQ ID NO: 29, where X is Val, Ala, Ser, or Pro), WKXKXKXKXK (SEQ ID NO: 30, where X is Val, Ala, Ser, or Pro), KWKVKVKVKVKVKVK (SEQ ID NO: 31, where X is Val, A, Ser, or Pro), LLLLKKKKKKKKLLLL (SEQ ID NO: 32), VKVKVKVKVDPPTKVKVKVKV (SEQ ID NO: 33), VKVKVKVKVDPPTKVKTKVKV (SEQ ID NO: 34), KVKVKVKVKDPPSVKVKVKVK (SEQ ID NO: 35), VKVKVKVKVDPPSKVKVKVKV (SEQ ID NO: 36), VKVKVKTKVDPPTKVKTKVKV (SEQ ID NO: 37), QQKFxFQFEQQ (SEQ ID NO: 38, wherein x is Glu, Asp, or Asn), QQKFQxQFEQQ (SEQ ID NO: 39, wherein x is Trp or Tyr), and QQKFQFxFEQQ (SEQ ID NO: 40, wherein x is Glu, Asp, or Asn), and at least one PEG molecule conjugated to a terminus of the self-assembling peptide, (iii-b) wherein the PAS conjugate peptide comprises: a self-assembling peptide comprising a polypeptide having the amino acid sequence of QQKFQFQFEQQ (SEQ ID NO: 12), bXXXb (SEQ ID NO: 1, wherein X is independently any amino acid and b is independently any positively charged amino acid), FKFEFKFE (SEQ ID NO: 14), KFQFQFE (SEQ ID NO: 15), QQRFQFQFEQQ (SEQ ID NO: 16), QQRFQWQFEQQ (SEQ ID NO: 17), FEFEFKFKFEFEFKFK (SEQ ID NO: 18), QQRFEWEFEQQ (SEQ ID NO: 19), QQXFXWXFQQQ (SEQ ID NO: 20, where X is ornithine), FKFEFKFEFKFE (SEQ ID NO: 21), FKFQFKFQFKFQ (SEQ ID NO: 22), AEAKAEAKAEAKAEAK (SEQ ID NO: 23), AEAEAKAKAEAEAKAK (SEQ ID NO: 24), AEAEAEAEAKAKAKAK (SEQ ID NO: 25), RADARADARADARADA (SEQ ID NO: 26), RARADADARARADADA (SEQ ID NO: 27), SGRGYBLGGQGAGAAAAAGGAGQGGYGGLGSQG (SEQ ID NO: 28), EWEXEXEXEX (SEQ ID NO: 29, where X is Val, Ala, Ser, or Pro), WKXKXKXKXK (SEQ ID NO: 30, where XDocket No.028193-0048-WO01 / 8355 is Val, Ala, Ser, or Pro), KWKVKVKVKVKVKVK (SEQ ID NO: 31, where X is Val, A, Ser, or Pro), LLLLKKKKKKKKLLLL (SEQ ID NO: 32), VKVKVKVKVDPPTKVKVKVKV (SEQ ID NO: 33), VKVKVKVKVDPPTKVKTKVKV (SEQ ID NO: 34), KVKVKVKVKDPPSVKVKVKVK (SEQ ID NO: 35), VKVKVKVKVDPPSKVKVKVKV (SEQ ID NO: 36), VKVKVKTKVDPPTKVKTKVKV (SEQ ID NO: 37), QQKFxFQFEQQ (SEQ ID NO: 38, wherein x is Glu, Asp, or Asn), QKFQxQFEQQ (SEQ ID NO: 39, wherein x is Trp or Tyr), and QQKFQFxFEQQ (SEQ ID NO: 40, wherein x is Glu, Asp, or Asn); and at least one PAS peptide conjugated to a terminus of the self-assembling peptide, wherein the PAS peptide comprises a sequence of Pro-Ala-Ser or comprises the sequence of ASPAAPAPASPAAPAPSAPA (SEQ ID NO: 97), or a peptide having at least 80%, 85%, 90%, or 95% identity thereto, and (iv) wherein the plain self-assembling peptide comprises a polypeptide having the amino acid sequence of bXXXb (SEQ ID NO: 1, wherein X is independently any amino acid and b is independently any positively charged amino acid), QQKFQFQFEQQ (SEQ ID NO: 12), FKFEFKFE (SEQ ID NO: 14), KFQFQFE (SEQ ID NO: 15), QQRFQFQFEQQ (SEQ ID NO: 16), QQRFQWQFEQQ (SEQ ID NO: 17), FEFEFKFKFEFEFKFK (SEQ ID NO: 18), QQRFEWEFEQQ (SEQ ID NO: 19), QQXFXWXFQQQ (SEQ ID NO: 20, where X is ornithine), FKFEFKFEFKFE (SEQ ID NO: 21), FKFQFKFQFKFQ (SEQ ID NO: 22), AEAKAEAKAEAKAEAK (SEQ ID NO: 23), AEAEAKAKAEAEAKAK (SEQ ID NO: 24), AEAEAEAEAKAKAKAK (SEQ ID NO: 25), RADARADARADARADA (SEQ ID NO: 26), RARADADARARADADA (SEQ ID NO: 27), SGRGYBLGGQGAGAAAAAGGAGQGGYGGLGSQG (SEQ ID NO: 28), EWEXEXEXEX (SEQ ID NO: 29, where X is Val, Ala, Ser, or Pro), WKXKXKXKXK (SEQ ID NO: 30, where X is Val, Ala, Ser, or Pro), KWKVKVKVKVKVKVK (SEQ ID NO: 31, where X is Val, A, Ser, or Pro), LLLLKKKKKKKKLLLL (SEQ ID NO: 32), VKVKVKVKVDPPTKVKVKVKV (SEQ ID NO: 33), VKVKVKVKVDPPTKVKTKVKV (SEQ ID NO: 34), KVKVKVKVKDPPSVKVKVKVK (SEQ ID NO: 35), VKVKVKVKVDPPSKVKVKVKV (SEQ ID NO: 36), VKVKVKTKVDPPTKVKTKVKV (SEQ ID NO: 37), QQKFxFQFEQQ (SEQ ID NO: 38, wherein x is Glu, Asp, or Asn), QQKFQxQFEQQ (SEQ ID NO: 39, wherein x is Trp or Tyr), or QQKFQFxFEQQ (SEQ ID NO: 40, wherein x is Glu, Asp, or Asn). [000157] Clause 27. The nanofiber of clause 26, wherein the cyclohexylalanine comprises D-alanine. [000158] Clause 28. The nanofiber of clause 26 or 27, wherein the T-cell epitope- conjugate peptide further comprises a linker between the T-cell epitope and the self- assembling peptide.Docket No.028193-0048-WO01 / 8355 [000159] Clause 29. The nanofiber of clause 28, wherein the linker comprises an aminoacid sequence selected from SEQ ID NO: 83 (SGSG), SEQ ID NO: 84 ((Ser-Gly)2), SEQ IDNO: 85 (CCCCSGSG), SEQ ID NO: 86 (Gnwherein n is an integer from 1 to 10), SEQ IDNO: 87 (GSGS), SEQ ID NO: 88 (SSSS), SEQ ID NO: 89 (GGGS), SEQ ID NO: 90 (GGC),SEQ ID NO: 91 ((GGC)8), SEQ ID NO: 92 ((G4S)3), SEQ ID NO: 93 (KSGSG), SEQ ID NO:94 (KKSGSG), SEQ ID NO: 95 (EAAAK)2, and SEQ ID NO: 96 (GGAAY).[000160] Clause 30. The nanofiber of any one of clauses 26-29, wherein the nanofiber comprises allergen conjugate peptides and plain self-assembling peptides. [000161] Clause 31. The nanofiber of any one of clauses 26-30, wherein adjacent allergen epitopes in the nanofiber have more than about 0.22 nm, more than about 0.88 nm, about 0.9 nm to about 22 nm, about 10 nm to about 20 nm, more than about 22 nm, less than about 440 nm, more than about 0.22 nm to less than about 440 nm, more than about 0.88 nm to less than about 440 nm, more than about 22 nm to less than about 440 nm, about 25 nm to about 30 nm, or about 28 nm between them. [000162] Clause 32. The nanofiber of any one of clauses 26-31, wherein more than about 0.05%, less than about 1%, less than about 25%, less than about 20%, at least about 0.05%, at least about 0.1%, at least about 0.2%, at least about 0.3%, at least about 0.4%, at least about 0.5%, at least about 0.6%, at least about 0.7%, at least about 0.8%, at least about 0.9%, at least about 1%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% of the peptides in the nanofiber are allergen conjugate peptides. [000163] Clause 33. The nanofiber of any one of clauses 26-32, wherein the nanofiber comprises about 0.2-1% allergen conjugate peptides and 99-99.8% plain self-assembling peptides, or about 0.5% allergen conjugate peptides and 99.5% plain self-assembling peptides. [000164] Clause 34. The nanofiber of any one of clauses 25-33, wherein the nanofiber comprises a plurality of the same or different allergen epitopes.Docket No.028193-0048-WO01 / 8355 [000165] Clause 35. The nanofiber of any one of clauses 26-34, wherein the nanofiber comprises a combination of 10 to 10,000, or 100 to 10,000 peptides comprising allergen conjugate peptides and plain self-assembling peptides. [000166] Clause 36. The nanofiber of any one of clauses 26-35, wherein the nanofiber comprises a combination of 10 to 10,000, or 100 to 10,000 peptides comprising allergen conjugate peptides, T-cell epitope-conjugate peptides, and plain self-assembling peptides. [000167] Clause 37. The nanofiber of any one of clauses 26-35, wherein the nanofiber comprises a combination of 10 to 10,000, or 100 to 10,000 peptides comprising allergen conjugate peptides, T-cell epitope-conjugate peptides, PAS conjugate peptides or PEG conjugate peptides, and plain self-assembling peptides. [000168] Clause 38. The nanofiber of any one of clauses 26-35, wherein the nanofiber comprises a combination of 10 to 10,000, or 100 to 10,000 peptides comprising allergen conjugate peptides, PAS conjugate peptides or PEG conjugate peptides, and plain self- assembling peptides. [000169] Clause 39. The nanofiber of any one of clauses 26-38, wherein at least about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, or 97.5% of the peptides in the nanofiber are allergen conjugate peptides. [000170] Clause 40. The nanofiber of any one of clauses 26-39, wherein at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, or 15% of the peptides in the nanofiber are T-cell epitope-conjugate peptides. [000171] Clause 41. The nanofiber of any one of clauses 26-40, wherein the allergen conjugate peptide and the T-cell epitope-conjugate peptide are present in the nanofiber at a ratio of about 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 12:1, 14:1, 16:1, 18:1, 20:1, 22:1, 24:1, 26:1, 28:1, 30:1, 32:1, 34:1, 36:1, 38:1, or 40:1. [000172] Clause 42. The nanofiber of any one of clauses 25-41, wherein the self- assembling peptide forms a fibril including beta-sheet structures or a fibril having a coiled coil structure. [000173] Clause 43. The nanofiber of any one of clauses 25-41, wherein the self- assembling peptide forms a fibril having a structure of a helical filament formed around a central axis.Docket No.028193-0048-WO01 / 8355 [000174] Clause 44. The nanofiber of any one of clauses 25-43, wherein the N-terminus of each self-assembling peptide is positioned at the exterior of the helical filament. [000175] Clause 45. The nanofiber of any one of clauses 25-44, wherein the allergen epitopes are exposed on the exterior surface of the nanofiber. [000176] Clause 46. The nanofiber of any one of clauses 25-45, wherein the nanofiber is about 5-30 nm in width. [000177] Clause 47. The nanofiber of any one of clauses 25-46, wherein the nanofiber is about 100 nm to 1 μm, 100 nm to 2 μm, 100 nm to 3 μm, 100 nm to 4 μm, or 100 nm to 5 μm in length. [000178] Clause 48. A pharmaceutical composition comprising: (a) the allergen conjugate peptide of any one of clauses 1-24 or the nanofiber of any one of clauses 25-47; and (b) a pharmaceutically acceptable carrier, diluent, and / or excipient. [000179] Clause 49. The pharmaceutical composition of clause 48, further comprising: (c) an adjuvant selected from cyclic-di-AMP, CpG, cyclic GMP-AMP (cGAMP), cholera toxin B subunit (CTB), retinoic acid, heat labile toxin B subunit, alum, MF59, 3M-052, iscomatrix, squalene-based adjuvants, AS01, AS03, or AS04, or a combination thereof. [000180] Clause 50. A method of treating an allergy, the method comprising administering to a subject a therapeutically effective amount of the allergen conjugate peptide of any one of clauses 1-24, or the nanofiber of any one of clauses 25-47, or the pharmaceutical composition of any one of clauses 48-49. [000181] Clause 51. A method of reducing inflammation in a subject, the method comprising administering to the subject a therapeutically effective amount of the allergen conjugate peptide of any one of clauses 1-24, or the nanofiber of any one of clauses 25-47, or the pharmaceutical composition of any one of clauses 48-49. [000182] Clause 52. The method of any one of clauses 50-51, wherein the allergen conjugate peptide, the nanofiber, or the pharmaceutical composition as administered subcutaneously or sublingually or orally. [000183] Clause 53. The method of any one of clauses 50-52, wherein the allergen does not leak into the vascular compartment or bloodstream.Docket No.028193-0048-WO01 / 8355 [000184] Clause 54. The method of any one of clauses 50-53, wherein the subject forms allergen-specific IgG antibodies. [000185] Clause 55. The method of clause 54, wherein the allergen-specific IgG antibodies are detected in the subject for at least 20 weeks, at least 21 weeks, at least 22 weeks, at least 23 weeks, at least 24 weeks, at least 25 weeks, at least 30 weeks, at least 35 weeks, at least 40 weeks, at least 45 weeks, at least 50 weeks, at least 51 weeks, or at least 52 weeks. [000186] Clause 56. The method of any one of clauses 50-55, wherein the subject becomes desensitized to the allergen. [000187] Clause 57. The method of any one of clauses 50-56, wherein the subject has a reduced Type I hypersensitivity reaction to the allergen. [000188] Clause 58. The method of any one of clauses 50-57, wherein activation of bone marrow-derived mast cells (BMMC) in the subject is reduced. SEQUENCES SEQ ID NO: 1 bXXXb wherein X is independently any amino acid, and b is independently any positively charged amino acid. SEQ ID NO: 2 RAYAR SEQ ID NO: 3 KAYAK SEQ ID NO: 4 RXXXR wherein X is any amino acid. SEQ ID NO: 5ZnbXXXbZmwherein b is independently any positively charged amino acid, Z is independently any amino acid, X is independently any amino acid, n is an integer from 0 to 20, and m is an integer from 0 to 20. SEQ ID NO: 6 Coil29 QARILEADAEILRAYARILEAHAEILRAQ SEQ ID NO: 7 QAKILEADAEILKAYAKILEAHAEILKAQDocket No.028193-0048-WO01 / 8355 SEQ ID NO: 8 Coil23 ADAEILRAYARILEAHAEILRAQ SEQ ID NO: 9Ac-QARILEADAEILRAYARILEAHAEILRAQ-NH2SEQ ID NO: 10Ac-QAKILEADAEILKAYAKILEAHAEILKAQ-NH2SEQ ID NO: 11Ac-ADAEILRAYARILEAHAEILRAQ-NH2SEQ ID NO: 12 Q11 QQKFQFQFEQQ SEQ ID NO: 13Ac-QQKFQFQFEQQ-NH2FKFEFKFE (SEQ ID NO: 14) KFQFQFE (SEQ ID NO: 15) QQRFQFQFEQQ (SEQ ID NO: 16) QQRFQWQFEQQ (SEQ ID NO: 17) FEFEFKFKFEFEFKFK (SEQ ID NO: 18) QQRFEWEFEQQ (SEQ ID NO: 19) QQXFXWXFQQQ (SEQ ID NO: 20, where X is ornithine) FKFEFKFEFKFE (SEQ ID NO: 21) FKFQFKFQFKFQ (SEQ ID NO: 22) AEAKAEAKAEAKAEAK (SEQ ID NO: 23) AEAEAKAKAEAEAKAK (SEQ ID NO: 24) AEAEAEAEAKAKAKAK (SEQ ID NO: 25) RADARADARADARADA (SEQ ID NO: 26) RARADADARARADADA (SEQ ID NO: 27) SGRGYBLGGQGAGAAAAAGGAGQGGYGGLGSQG (SEQ ID NO: 28) EWEXEXEXEX (SEQ ID NO: 29, where X is Val, Ala, Ser, or Pro) WKXKXKXKXK (SEQ ID NO: 30, where X is Val, Ala, Ser, or Pro)Docket No.028193-0048-WO01 / 8355 KWKVKVKVKVKVKVK (SEQ ID NO: 31, where X is Val, A, Ser, or Pro) LLLLKKKKKKKKLLLL (SEQ ID NO: 32) VKVKVKVKVDPPTKVKVKVKV (SEQ ID NO: 33) VKVKVKVKVDPPTKVKTKVKV (SEQ ID NO: 34) KVKVKVKVKDPPSVKVKVKVK (SEQ ID NO: 35) VKVKVKVKVDPPSKVKVKVKV (SEQ ID NO: 36) VKVKVKTKVDPPTKVKTKVKV (SEQ ID NO: 37) QQKFxFQFEQQ (SEQ ID NO: 38, wherein x is Glu, Asp, or Asn) QQKFQxQFEQQ (SEQ ID NO: 39, wherein x is Trp or Tyr) QQKFQFxFEQQ (SEQ ID NO: 40, wherein x is Glu, Asp, or Asn) VEVKVEVKV (SEQ ID NO: 41) VEVKVEVKVEVK (SEQ ID NO: 42) VVVAAAEEE (SEQ ID NO: 43) VEVEVEVEVEVEVEVEVEVE (SEQ ID NO: 44) CGNKRTRGC (SEQ ID NO: 45) VKVKVKVKVDPPTKVEVKVKV (SEQ ID NO: 46) LRKKLGKA (SEQ ID NO: 47) VVVVVVKK (SEQ ID NO: 48) AEAKAEAKAEAKAEAK (SEQ ID NO: 49) AEAKAEAK (SEQ ID NO: 50) AEAEAEAEAKAK (SEQ ID NO: 51) AEAEAKAK (SEQ ID NO: 52) AEAEAKAKAEAEAKAK (SEQ ID NO: 53) RADARADARADARADA (SEQ ID NO: 54) RADARGDARADARGDA (SEQ ID NO: 55) RADARADA (SEQ ID NO: 56) RARADADARARADADA (SEQ ID NO: 57)Docket No.028193-0048-WO01 / 8355 RARADADA (SEQ ID NO: 58) RARARARADADADADA (SEQ ID NO: 59) ADADADADARARARAR (SEQ ID NO: 60) DADADADARARARARA (SEQ ID NO: 61) RAEARAEARAEARAEA (SEQ ID NO: 62) RAEARAEA (SEQ ID NO: 63) KAKAKAKAEAEAEAEA (SEQ ID NO: 64) AEAEAEAEAKAKAKAK (SEQ ID NO: 65) KADAKADAKADAKADA (SEQ ID NO: 66) KADAKADA (SEQ ID NO: 67) AEAEAHAHAEAEAHAHA (SEQ ID NO: 68) AEAEAHAHA (SEQ ID NO: 69) HEHEHKHKHEHEHKHK (SEQ ID NO: 70) HEHEHKHK (SEQ ID NO: 71) FEFEFKFKFEFEFKFK (SEQ ID NO: 72) FEFKFEFK (SEQ ID NO: 73) LELELKLKLELELKLK (SEQ ID NO: 74) LELELKLK (SEQ ID NO: 75) KFDLKKDLKLDL (SEQ ID NO: 76) FKFEFKFF (SEQ ID NO: 77) FEFEFKFK (SEQ ID NO: 78) RFRFRFRFRFRFRFRFRFRF (SEQ ID NO: 79) SEQ ID NO: 80 PADRE molecule with C-terminal NH2 (amidation) NH2-aKXVAAWTLKAa-NH2 Wherein “X” comprises cyclohexylalanine and “a” comprises D-alanine. SEQ ID NO: 81 PADRE-Coil29 with C-terminal NH2(amidation) NH2-aKXVAAWTLKAa-SGSG-QARILEADAEILRAYARILEAHAEILRAQ-NH2 Wherein X = cyclohexyl alanine, a = D-alanine, C-terminal NH2indicates amidation.Docket No.028193-0048-WO01 / 8355 SEQ ID NO: 82 TAMRA-Coil29 with C-terminal NH2(amidation) TAMRA-SGSGEAAAKEAAAKSGSG -QARILEADAEILRAYARILEAHAEILRAQ-NH2 C-terminal NH2 indicates amidation. SEQ ID NO: 83 Linker SGSG SEQ ID NO: 84 Linker(Ser-Gly)2SEQ ID NO: 85 Linker CCCCSGSG SEQ ID NO: 86 LinkerGnwherein n is an integer from 1 to 10SEQ ID NO: 87 Linker GSGS SEQ ID NO: 88 Linker SSSS SEQ ID NO: 89 Linker GGGS SEQ ID NO: 90 Linker GGC SEQ ID NO: 91 Linker(GGC)8SEQ ID NO: 92 Linker(G4S)3SEQ ID NO: 93 Linker KSGSG SEQ ID NO: 94 Linker KKSGSGDocket No.028193-0048-WO01 / 8355 SEQ ID NO: 95 Linker(EAAAK)2SEQ ID NO: 96 Linker GGAAY SEQ ID NO: 97 PAS peptide ASPAAPAPASPAAPAPSAPA SEQ ID NO: 98 PAS peptideH2N-ASPAAPAPASPAAPAPSAPA-NH2SEQ ID NO; 99 PADRE molecule aKXVAAWTLKAa, wherein “X” comprises cyclohexylalanine and “a” comprises D-alanine SEQ ID NO: 100 VAC molecule QLVFNSISARALKAY QAEILRAYARILEAQ (SEQ ID NO: 101) QAEILRAYARILEAHAEILKAQ (SEQ ID NO: 102) QAEILRAYARILEADAKILEAHAEILKAQ (SEQ ID NO: 103) QAEILRAYARILEADAEILKAQAKILEAHAEILKAQ (SEQ ID NO: 104) SGSG-C-SGSG (SEQ ID NO: 105) AH1b (REREREEDWRQPREDWRRPS, SEQ ID NO: 106) AH3a (EDEYEYDEEDRRRGRGSRGR, SEQ ID NO: 107) AH7a (QEQDEYPYSRRGSRGRQPGE, SEQ ID NO: 108) AH1a (IDQIEKQAKDLAFPGSGE, SEQ ID NO: 109) AH2a (DPYSOSQDPYSOS, SEQ ID NO: 110, wherein O is hydroxyproline) AH2b (DPYSOSDRRGAGSS, SEQ ID NO: 111, wherein O is hydroxyproline) SEQ ID NO: 112 (OVA61-68)Q11 H2N-QQKFQFQFEQQ-SGSG-DKLPGFDG-Am (Am is amide) SEQ ID NO: 113 (PADRE)Q11 NH2-aKXVAAWTLKAa-SGSG-QQKFQFQFEQQ-AmDocket No.028193-0048-WO01 / 8355 (a is D-alanine; Am is amide) SEQ ID NO: 114 Biotin-OVA61-68 Biotin-SGSG-DKLPGFDG-Am (Am is amide) SEQ ID NO: 115 AH1a-Q11 peptide H2N-IDQIEKQAKDLAFPGSGE-SGSG-QQKFQFQFEQQ-Am (Am is amide) SEQ ID NO: 116 Q11-AH1b peptide H2N-QQKFQFQFEQQ-SGSG-REREREEDWRQPREDWRRPS-Am (Am is amide) SEQ ID NO: 117 AH2a-Q11 peptide H2N-DPYSOSQDPYSOS-SGSG-QQKFQFQFEQQ-Am *O is hydroxyproline (Hyp) and Am is amide. SEQ ID NO: 118 Q11-AH2b peptide H2N-QQKFQFQFEQQ-SGSG-DPYSOSDRRGAGSS-Am *O is hydroxyproline (Hyp) and Am is amide. SEQ ID NO: 119 Q11-AH3a peptide H2N-QQKFQFQFEQQ-SGSG-EDEYEYDEEDRRRGRGSRGR-Am (Am is amide) SEQ ID NO: 120 Q11-AH7a peptide H2N-QQKFQFQFEQQ-SGSG-QEQDEYPYSRRGSRGRQPGE-Am (Am is amide)

Claims

Docket No.028193-0048-WO01 / 8355 What is claimed is:

1. An allergen conjugate peptide comprising: (i) a self-assembling peptide comprising a polypeptide having the amino acid sequence of bXXXb (SEQ ID NO: 1, wherein X is independently any amino acid and b is independently any positively charged amino acid), QQKFQFQFEQQ (SEQ ID NO: 12), FKFEFKFE (SEQ ID NO: 14), KFQFQFE (SEQ ID NO: 15), QQRFQFQFEQQ (SEQ ID NO: 16), QQRFQWQFEQQ (SEQ ID NO: 17), FEFEFKFKFEFEFKFK (SEQ ID NO: 18), QQRFEWEFEQQ (SEQ ID NO: 19), QQXFXWXFQQQ (SEQ ID NO: 20, where X is ornithine), FKFEFKFEFKFE (SEQ ID NO: 21), FKFQFKFQFKFQ (SEQ ID NO: 22), AEAKAEAKAEAKAEAK (SEQ ID NO: 23), AEAEAKAKAEAEAKAK (SEQ ID NO: 24), AEAEAEAEAKAKAKAK (SEQ ID NO: 25), RADARADARADARADA (SEQ ID NO: 26), RARADADARARADADA (SEQ ID NO: 27), SGRGYBLGGQGAGAAAAAGGAGQGGYGGLGSQG (SEQ ID NO: 28), EWEXEXEXEX (SEQ ID NO: 29, where X is Val, Ala, Ser, or Pro), WKXKXKXKXK (SEQ ID NO: 30, where X is Val, Ala, Ser, or Pro), KWKVKVKVKVKVKVK (SEQ ID NO: 31, where X is Val, A, Ser, or Pro), LLLLKKKKKKKKLLLL (SEQ ID NO: 32), VKVKVKVKVDPPTKVKVKVKV (SEQ ID NO: 33), VKVKVKVKVDPPTKVKTKVKV (SEQ ID NO: 34), KVKVKVKVKDPPSVKVKVKVK (SEQ ID NO: 35), VKVKVKVKVDPPSKVKVKVKV (SEQ ID NO: 36), VKVKVKTKVDPPTKVKTKVKV (SEQ ID NO: 37), QQKFxFQFEQQ (SEQ ID NO: 38, wherein x is Glu, Asp, or Asn), QQKFQxQFEQQ (SEQ ID NO: 39, wherein x is Trp or Tyr), or QQKFQFxFEQQ (SEQ ID NO: 40, wherein x is Glu, Asp, or Asn); and (ii) at least one allergen epitope conjugated to a terminus of the self-assembling peptide.

2. The allergen conjugate peptide of claim 1, wherein the allergen epitope is a B cell allergen epitope.

3. The allergen conjugate peptide of claim 1 or 2, wherein the allergen epitope is IgE- reactive.

4. The allergen conjugate peptide of any one of claims 1-3, wherein the allergen epitope comprises a peanut allergen epitope.Docket No.028193-0048-WO01 / 8355 5. The allergen conjugate peptide of claim 4, wherein the peanut allergen epitope is from peanut allergen protein Ara h 1, Ara h 3, or Ara h 7, or Ara h 2.

6. The allergen conjugate peptide of claim 5, wherein the peanut allergen epitope comprises AH1b, AH3a, AH7a, AH1a, AH2a, or AH2b, or a combination thereof.

7. The allergen conjugate peptide of claim 6, wherein the peanut allergen epitope comprises AH1b, AH3a, or AH7a, or a combination thereof.

8. The allergen conjugate peptide of any one of claims 1-7, wherein each self- assembling peptide forms a beta sheet and comprises a polypeptide having an amino acid sequence selected from QQKFQFQFEQQ (SEQ ID NO: 12), FKFEFKFE (SEQ ID NO: 14), KFQFQFE (SEQ ID NO: 15), QQRFQFQFEQQ (SEQ ID NO: 16), QQRFQWQFEQQ (SEQ ID NO: 17), FEFEFKFKFEFEFKFK (SEQ ID NO: 18), QQRFEWEFEQQ (SEQ ID NO: 19), QQXFXWXFQQQ (SEQ ID NO: 20, where X is ornithine), FKFEFKFEFKFE (SEQ ID NO: 21), FKFQFKFQFKFQ (SEQ ID NO: 22), AEAKAEAKAEAKAEAK (SEQ ID NO: 23), AEAEAKAKAEAEAKAK (SEQ ID NO: 24), AEAEAEAEAKAKAKAK (SEQ ID NO: 25), RADARADARADARADA (SEQ ID NO: 26), RARADADARARADADA (SEQ ID NO: 27), SGRGYBLGGQGAGAAAAAGGAGQGGYGGLGSQG (SEQ ID NO: 28), EWEXEXEXEX (SEQ ID NO: 29, where X is Val, Ala, Ser, or Pro), WKXKXKXKXK (SEQ ID NO: 30, where X is Val, Ala, Ser, or Pro), KWKVKVKVKVKVKVK (SEQ ID NO: 31, where X is Val, A, Ser, or Pro), LLLLKKKKKKKKLLLL (SEQ ID NO: 32), VKVKVKVKVDPPTKVKVKVKV (SEQ ID NO: 33), VKVKVKVKVDPPTKVKTKVKV (SEQ ID NO: 34), KVKVKVKVKDPPSVKVKVKVK (SEQ ID NO: 35), VKVKVKVKVDPPSKVKVKVKV (SEQ ID NO: 36), VKVKVKTKVDPPTKVKTKVKV (SEQ ID NO: 37), QQKFxFQFEQQ (SEQ ID NO: 38, wherein x is Glu, Asp, or Asn), QQKFQxQFEQQ (SEQ ID NO: 39, wherein x is Trp or Tyr), and QQKFQFxFEQQ (SEQ ID NO: 40, wherein x is Glu, Asp, or Asn).

9. The allergen conjugate peptide of any one of claims 1-7, wherein each self- assembling peptide forms an alpha-helix and comprises a polypeptide having an amino acid sequence of bXXXb (SEQ ID NO: 1), wherein X is independently any amino acid and b is independently any positively charged amino acid.

10. The allergen conjugate peptide according to claim 8 or 9, wherein the self- assembling peptide comprises the sequence QQKFQFQFEQQ (SEQ ID NO: 12) or Ac-Docket No.028193-0048-WO01 / 8355QQKFQFQFEQQ-NH2(SEQ ID NO: 13) or bXXXb (SEQ ID NO: 1, wherein X isindependently any amino acid and b is independently any positively charged amino acid).

11. The allergen conjugate peptide of any one of claims 1-7 and 9-10, wherein b is independently selected from Arg and Lys.

12. The allergen conjugate peptide of any one of claims 1-7 and 9-11, wherein bXXXb (SEQ ID NO: 1) is RAYAR (SEQ ID NO: 2) or KAYAK (SEQ ID NO: 3).

13. The allergen conjugate peptide of any one of claims 1-7 and 9-12, wherein the self-assembling peptide comprises an amino acid sequence of ZnbXXXbZm(SEQ ID NO: 5),wherein b is independently any positively charged amino acid, Z is independently any amino acid, X is independently any amino acid, n is an integer from 0 to 20, and m is an integer from 0 to 20.

14. The allergen conjugate peptide of claim 13, wherein the self-assembling peptide comprises an amino acid sequence selected from QARILEADAEILRAYARILEAHAEILRAQ (Coil29, SEQ ID NO: 6), or QAKILEADAEILKAYAKILEAHAEILKAQ (SEQ ID NO: 7), or ADAEILRAYARILEAHAEILRAQ (SEQ ID NO: 8), or Ac-QARILEADAEILRAYARILEAHAEILRAQ-NH2(SEQ ID NO: 9), or Ac-QAKILEADAEILKAYAKILEAHAEILKAQ-NH2(SEQ ID NO: 10), or Ac-ADAEILRAYARILEAHAEILRAQ-NH2(SEQ ID NO: 11), or QAEILRAYARILEAQ (SEQ IDNO: 101), or QAEILRAYARILEAHAEILKAQ (SEQ ID NO: 102), or QAEILRAYARILEADAKILEAHAEILKAQ (SEQ ID NO: 103), or QAEILRAYARILEADAEILKAQAKILEAHAEILKAQ (SEQ ID NO: 104).

15. The allergen conjugate peptide of any one of claims 1-14, wherein the self- assembling peptide forms a beta sheet and the at least one allergen epitope is attached to the C-terminus or the N-terminus of the self-assembling peptide or a combination thereof, or wherein the self-assembling peptide forms an alpha-helix and the at least one allergen epitope is attached to the N-terminus of the self-assembling peptide.

16. The allergen conjugate peptide of any one of claims 1-15, wherein 1 to 10 allergen epitopes are attached to the C-terminus or the N-terminus of the self-assembling peptide.Docket No.028193-0048-WO01 / 8355 17. The allergen conjugate peptide of any one of claims 1-8 and 15-16, wherein the self- assembling peptide forms a beta sheet and the allergen conjugate peptide further comprises: (iii) a PEG molecule or a PAS peptide conjugated to the self-assembling peptide.

18. The allergen conjugate peptide of claim 17, wherein the PAS peptide comprises a sequence of Pro-Ala-Ser or comprises the sequence of ASPAAPAPASPAAPAPSAPA (SEQ ID NO: 97), or a peptide having at least 80%, 85%, 90%, or 95% identity thereto.

19. The allergen conjugate peptide of claim 17, wherein the PEG molecule comprises PEG-2000.

20. The allergen conjugate of any one of claims 17-19, wherein the PEG molecule or the PAS peptide is conjugated to the self-assembling peptide at the same or the opposite terminus from wherein the allergen epitope is attached.

21. The allergen conjugate peptide of any one of claims 1-20, further comprising: (iv) at least one linker.

22. The allergen conjugate peptide of claim 21, wherein the at least one linker is between the at least one allergen epitope and the self-assembling peptide.

23. The allergen conjugate peptide of claim 21, wherein the at least one linker comprises a first linker between the at least one allergen epitope and the self-assembling peptide, and a second linker between the PEG molecule or the PAS peptide and the self-assembling peptide.

24. The allergen conjugate peptide of any one of claims 21-23, wherein the at least onelinker comprises SEQ ID NO: 83 (SGSG), SEQ ID NO: 84 ((Ser-Gly)2), SEQ ID NO: 85(CCCCSGSG), SEQ ID NO: 86 (Gnwherein n is an integer from 1 to 10), SEQ ID NO: 87(GSGS), SEQ ID NO: 88 (SSSS), SEQ ID NO: 89 (GGGS), SEQ ID NO: 90 (GGC), SEQ IDNO: 91 ((GGC)8), SEQ ID NO: 92 ((G4S)3), SEQ ID NO: 93 (KSGSG), SEQ ID NO: 94(KKSGSG), SEQ ID NO: 95 (EAAAK)2, or SEQ ID NO: 96 (GGAAY).Docket No.028193-0048-WO01 / 8355 25. A nanofiber comprising a plurality of the allergen conjugate peptide of any one of claims 1-24, wherein the plurality of allergen conjugate peptides self-assemble into the nanofiber.

26. A nanofiber comprising: (i) at least one allergen conjugate peptide of any one of claims 1-24; and at least one of a peptide selected from a (ii) T-cell epitope-conjugate peptide, a (iii-a) PEG conjugate peptide, a (iii-b) PAS conjugate peptide, and a (iv) plain self-assembling peptide, or a combination thereof, (ii) wherein the T-cell epitope-conjugate peptide comprises: a self-assembling peptide comprising a polypeptide having the amino acid sequence of QQKFQFQFEQQ (SEQ ID NO: 12), bXXXb (SEQ ID NO: 1, wherein X is independently any amino acid and b is independently any positively charged amino acid), FKFEFKFE (SEQ ID NO: 14), KFQFQFE (SEQ ID NO: 15), QQRFQFQFEQQ (SEQ ID NO: 16), QQRFQWQFEQQ (SEQ ID NO: 17), FEFEFKFKFEFEFKFK (SEQ ID NO: 18), QQRFEWEFEQQ (SEQ ID NO: 19), QQXFXWXFQQQ (SEQ ID NO: 20, where X is ornithine), FKFEFKFEFKFE (SEQ ID NO: 21), FKFQFKFQFKFQ (SEQ ID NO: 22), AEAKAEAKAEAKAEAK (SEQ ID NO: 23), AEAEAKAKAEAEAKAK (SEQ ID NO: 24), AEAEAEAEAKAKAKAK (SEQ ID NO: 25), RADARADARADARADA (SEQ ID NO: 26), RARADADARARADADA (SEQ ID NO: 27), SGRGYBLGGQGAGAAAAAGGAGQGGYGGLGSQG (SEQ ID NO: 28), EWEXEXEXEX (SEQ ID NO: 29, where X is Val, Ala, Ser, or Pro), WKXKXKXKXK (SEQ ID NO: 30, where X is Val, Ala, Ser, or Pro), KWKVKVKVKVKVKVK (SEQ ID NO: 31, where X is Val, A, Ser, or Pro), LLLLKKKKKKKKLLLL (SEQ ID NO: 32), VKVKVKVKVDPPTKVKVKVKV (SEQ ID NO: 33), VKVKVKVKVDPPTKVKTKVKV (SEQ ID NO: 34), KVKVKVKVKDPPSVKVKVKVK (SEQ ID NO: 35), VKVKVKVKVDPPSKVKVKVKV (SEQ ID NO: 36), VKVKVKTKVDPPTKVKTKVKV (SEQ ID NO: 37), QQKFxFQFEQQ (SEQ ID NO: 38, wherein x is Glu, Asp, or Asn), QQKFQxQFEQQ (SEQ ID NO: 39, wherein x is Trp or Tyr), and QQKFQFxFEQQ (SEQ ID NO: 40, wherein x is Glu, Asp, or Asn); and at least one T-cell epitope conjugated to a terminus of the self-assembling peptide, wherein the at least one T-cell epitope is selected from PADRE and VAC, and wherein PADRE comprises a polypeptide having the amino acid sequence of aKXVAAWTLKAa (SEQ ID NO: 99, wherein “X” comprises cyclohexylalanine and “a” comprises D-alanine), and wherein VAC comprises a polypeptide having the amino acid sequence of QLVFNSISARALKAY (SEQ ID NO: 100),Docket No.028193-0048-WO01 / 8355 (iii-a) wherein the PEG conjugate peptide comprises: a self-assembling peptide comprising a polypeptide having the amino acid sequence of QQKFQFQFEQQ (SEQ ID NO: 12), bXXXb (SEQ ID NO: 1, wherein X is independently any amino acid and b is independently any positively charged amino acid), FKFEFKFE (SEQ ID NO: 14), KFQFQFE (SEQ ID NO: 15), QQRFQFQFEQQ (SEQ ID NO: 16), QQRFQWQFEQQ (SEQ ID NO: 17), FEFEFKFKFEFEFKFK (SEQ ID NO: 18), QQRFEWEFEQQ (SEQ ID NO: 19), QQXFXWXFQQQ (SEQ ID NO: 20, where X is ornithine), FKFEFKFEFKFE (SEQ ID NO: 21), FKFQFKFQFKFQ (SEQ ID NO: 22), AEAKAEAKAEAKAEAK (SEQ ID NO: 23), AEAEAKAKAEAEAKAK (SEQ ID NO: 24), AEAEAEAEAKAKAKAK (SEQ ID NO: 25), RADARADARADARADA (SEQ ID NO: 26), RARADADARARADADA (SEQ ID NO: 27), SGRGYBLGGQGAGAAAAAGGAGQGGYGGLGSQG (SEQ ID NO: 28), EWEXEXEXEX (SEQ ID NO: 29, where X is Val, Ala, Ser, or Pro), WKXKXKXKXK (SEQ ID NO: 30, where X is Val, Ala, Ser, or Pro), KWKVKVKVKVKVKVK (SEQ ID NO: 31, where X is Val, A, Ser, or Pro), LLLLKKKKKKKKLLLL (SEQ ID NO: 32), VKVKVKVKVDPPTKVKVKVKV (SEQ ID NO: 33), VKVKVKVKVDPPTKVKTKVKV (SEQ ID NO: 34), KVKVKVKVKDPPSVKVKVKVK (SEQ ID NO: 35), VKVKVKVKVDPPSKVKVKVKV (SEQ ID NO: 36), VKVKVKTKVDPPTKVKTKVKV (SEQ ID NO: 37), QQKFxFQFEQQ (SEQ ID NO: 38, wherein x is Glu, Asp, or Asn), QQKFQxQFEQQ (SEQ ID NO: 39, wherein x is Trp or Tyr), and QQKFQFxFEQQ (SEQ ID NO: 40, wherein x is Glu, Asp, or Asn), and at least one PEG molecule conjugated to a terminus of the self-assembling peptide, (iii-b) wherein the PAS conjugate peptide comprises: a self-assembling peptide comprising a polypeptide having the amino acid sequence of QQKFQFQFEQQ (SEQ ID NO: 12), bXXXb (SEQ ID NO: 1, wherein X is independently any amino acid and b is independently any positively charged amino acid), FKFEFKFE (SEQ ID NO: 14), KFQFQFE (SEQ ID NO: 15), QQRFQFQFEQQ (SEQ ID NO: 16), QQRFQWQFEQQ (SEQ ID NO: 17), FEFEFKFKFEFEFKFK (SEQ ID NO: 18), QQRFEWEFEQQ (SEQ ID NO: 19), QQXFXWXFQQQ (SEQ ID NO: 20, where X is ornithine), FKFEFKFEFKFE (SEQ ID NO: 21), FKFQFKFQFKFQ (SEQ ID NO: 22), AEAKAEAKAEAKAEAK (SEQ ID NO: 23), AEAEAKAKAEAEAKAK (SEQ ID NO: 24), AEAEAEAEAKAKAKAK (SEQ ID NO: 25), RADARADARADARADA (SEQ ID NO: 26), RARADADARARADADA (SEQ ID NO: 27), SGRGYBLGGQGAGAAAAAGGAGQGGYGGLGSQG (SEQ ID NO: 28), EWEXEXEXEX (SEQ ID NO: 29, where X is Val, Ala, Ser, or Pro), WKXKXKXKXK (SEQ ID NO: 30, where X is Val, Ala, Ser, or Pro), KWKVKVKVKVKVKVK (SEQ IDDocket No.028193-0048-WO01 / 8355 NO: 31, where X is Val, A, Ser, or Pro), LLLLKKKKKKKKLLLL (SEQ ID NO: 32), VKVKVKVKVDPPTKVKVKVKV (SEQ ID NO: 33), VKVKVKVKVDPPTKVKTKVKV (SEQ ID NO: 34), KVKVKVKVKDPPSVKVKVKVK (SEQ ID NO: 35), VKVKVKVKVDPPSKVKVKVKV (SEQ ID NO: 36), VKVKVKTKVDPPTKVKTKVKV (SEQ ID NO: 37), QQKFxFQFEQQ (SEQ ID NO: 38, wherein x is Glu, Asp, or Asn), QQKFQxQFEQQ (SEQ ID NO: 39, wherein x is Trp or Tyr), and QQKFQFxFEQQ (SEQ ID NO: 40, wherein x is Glu, Asp, or Asn); and at least one PAS peptide conjugated to a terminus of the self-assembling peptide, wherein the PAS peptide comprises a sequence of Pro-Ala-Ser or comprises the sequence of ASPAAPAPASPAAPAPSAPA (SEQ ID NO: 97), or a peptide having at least 80%, 85%, 90%, or 95% identity thereto, and (iv) wherein the plain self-assembling peptide comprises a polypeptide having the amino acid sequence of bXXXb (SEQ ID NO: 1, wherein X is independently any amino acid and b is independently any positively charged amino acid), QQKFQFQFEQQ (SEQ ID NO: 12), FKFEFKFE (SEQ ID NO: 14), KFQFQFE (SEQ ID NO: 15), QQRFQFQFEQQ (SEQ ID NO: 16), QQRFQWQFEQQ (SEQ ID NO: 17), FEFEFKFKFEFEFKFK (SEQ ID NO: 18), QQRFEWEFEQQ (SEQ ID NO: 19), QQXFXWXFQQQ (SEQ ID NO: 20, where X is ornithine), FKFEFKFEFKFE (SEQ ID NO: 21), FKFQFKFQFKFQ (SEQ ID NO: 22), AEAKAEAKAEAKAEAK (SEQ ID NO: 23), AEAEAKAKAEAEAKAK (SEQ ID NO: 24), AEAEAEAEAKAKAKAK (SEQ ID NO: 25), RADARADARADARADA (SEQ ID NO: 26), RARADADARARADADA (SEQ ID NO: 27), SGRGYBLGGQGAGAAAAAGGAGQGGYGGLGSQG (SEQ ID NO: 28), EWEXEXEXEX (SEQ ID NO: 29, where X is Val, Ala, Ser, or Pro), WKXKXKXKXK (SEQ ID NO: 30, where X is Val, Ala, Ser, or Pro), KWKVKVKVKVKVKVK (SEQ ID NO: 31, where X is Val, A, Ser, or Pro), LLLLKKKKKKKKLLLL (SEQ ID NO: 32), VKVKVKVKVDPPTKVKVKVKV (SEQ ID NO: 33), VKVKVKVKVDPPTKVKTKVKV (SEQ ID NO: 34), KVKVKVKVKDPPSVKVKVKVK (SEQ ID NO: 35), VKVKVKVKVDPPSKVKVKVKV (SEQ ID NO: 36), VKVKVKTKVDPPTKVKTKVKV (SEQ ID NO: 37), QQKFxFQFEQQ (SEQ ID NO: 38, wherein x is Glu, Asp, or Asn), QQKFQxQFEQQ (SEQ ID NO: 39, wherein x is Trp or Tyr), or QQKFQFxFEQQ (SEQ ID NO: 40, wherein x is Glu, Asp, or Asn).

27. The nanofiber of claim 26, wherein the cyclohexylalanine comprises D-alanine.

28. The nanofiber of claim 26 or 27, wherein the T-cell epitope-conjugate peptide further comprises a linker between the T-cell epitope and the self-assembling peptide.Docket No.028193-0048-WO01 / 8355 29. The nanofiber of claim 28, wherein the linker comprises an amino acid sequenceselected from SEQ ID NO: 83 (SGSG), SEQ ID NO: 84 ((Ser-Gly)2), SEQ ID NO: 85(CCCCSGSG), SEQ ID NO: 86 (Gnwherein n is an integer from 1 to 10), SEQ ID NO: 87(GSGS), SEQ ID NO: 88 (SSSS), SEQ ID NO: 89 (GGGS), SEQ ID NO: 90 (GGC), SEQ IDNO: 91 ((GGC)8), SEQ ID NO: 92 ((G4S)3), SEQ ID NO: 93 (KSGSG), SEQ ID NO: 94(KKSGSG), SEQ ID NO: 95 (EAAAK)2, and SEQ ID NO: 96 (GGAAY).

30. The nanofiber of any one of claims 26-29, wherein the nanofiber comprises allergen conjugate peptides and plain self-assembling peptides.

31. The nanofiber of any one of claims 26-30, wherein adjacent allergen epitopes in the nanofiber have more than about 0.22 nm, more than about 0.88 nm, about 0.9 nm to about 22 nm, about 10 nm to about 20 nm, more than about 22 nm, less than about 440 nm, more than about 0.22 nm to less than about 440 nm, more than about 0.88 nm to less than about 440 nm, more than about 22 nm to less than about 440 nm, about 25 nm to about 30 nm, or about 28 nm between them.

32. The nanofiber of any one of claims 26-31, wherein more than about 0.05%, less than about 1%, less than about 25%, less than about 20%, at least about 0.05%, at least about 0.1%, at least about 0.2%, at least about 0.3%, at least about 0.4%, at least about 0.5%, at least about 0.6%, at least about 0.7%, at least about 0.8%, at least about 0.9%, at least about 1%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% of the peptides in the nanofiber are allergen conjugate peptides.

33. The nanofiber of any one of claims 26-32, wherein the nanofiber comprises about 0.2-1% allergen conjugate peptides and 99-99.8% plain self-assembling peptides, or about 0.5% allergen conjugate peptides and 99.5% plain self-assembling peptides.

34. The nanofiber of any one of claims 25-33, wherein the nanofiber comprises a plurality of the same or different allergen epitopes.Docket No.028193-0048-WO01 / 8355 35. The nanofiber of any one of claims 26-34, wherein the nanofiber comprises a combination of 10 to 10,000, or 100 to 10,000 peptides comprising allergen conjugate peptides and plain self-assembling peptides.

36. The nanofiber of any one of claims 26-35, wherein the nanofiber comprises a combination of 10 to 10,000, or 100 to 10,000 peptides comprising allergen conjugate peptides, T-cell epitope-conjugate peptides, and plain self-assembling peptides.

37. The nanofiber of any one of claims 26-35, wherein the nanofiber comprises a combination of 10 to 10,000, or 100 to 10,000 peptides comprising allergen conjugate peptides, T-cell epitope-conjugate peptides, PAS conjugate peptides or PEG conjugate peptides, and plain self-assembling peptides.

38. The nanofiber of any one of claims 26-35, wherein the nanofiber comprises a combination of 10 to 10,000, or 100 to 10,000 peptides comprising allergen conjugate peptides, PAS conjugate peptides or PEG conjugate peptides, and plain self-assembling peptides.

39. The nanofiber of any one of claims 26-38, wherein at least about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, or 97.5% of the peptides in the nanofiber are allergen conjugate peptides.

40. The nanofiber of any one of claims 26-39, wherein at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, or 15% of the peptides in the nanofiber are T-cell epitope-conjugate peptides.

41. The nanofiber of any one of claims 26-40, wherein the allergen conjugate peptide and the T-cell epitope-conjugate peptide are present in the nanofiber at a ratio of about 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 12:1, 14:1, 16:1, 18:1, 20:1, 22:1, 24:1, 26:1, 28:1, 30:1, 32:1, 34:1, 36:1, 38:1, or 40:

1.

42. The nanofiber of any one of claims 25-41, wherein the self-assembling peptide forms a fibril including beta-sheet structures or a fibril having a coiled coil structure.

43. The nanofiber of any one of claims 25-41, wherein the self-assembling peptide forms a fibril having a structure of a helical filament formed around a central axis.Docket No.028193-0048-WO01 / 8355 44. The nanofiber of any one of claims 25-43, wherein the N-terminus of each self- assembling peptide is positioned at the exterior of the helical filament.

45. The nanofiber of any one of claims 25-44, wherein the allergen epitopes are exposed on the exterior surface of the nanofiber.

46. The nanofiber of any one of claims 25-45, wherein the nanofiber is about 5-30 nm in width.

47. The nanofiber of any one of claims 25-46, wherein the nanofiber is about 100 nm to 1 μm, 100 nm to 2 μm, 100 nm to 3 μm, 100 nm to 4 μm, or 100 nm to 5 μm in length.

48. A pharmaceutical composition comprising: (a) the allergen conjugate peptide of any one of claims 1-24 or the nanofiber of any one of claims 25-47; and (b) a pharmaceutically acceptable carrier, diluent, and / or excipient.

49. The pharmaceutical composition of claim 48, further comprising: (c) an adjuvant selected from cyclic-di-AMP, CpG, cyclic GMP-AMP (cGAMP), cholera toxin B subunit (CTB), retinoic acid, heat labile toxin B subunit, alum, MF59, 3M-052, iscomatrix, squalene- based adjuvants, AS01, AS03, or AS04, or a combination thereof.

50. A method of treating an allergy, the method comprising administering to a subject a therapeutically effective amount of the allergen conjugate peptide of any one of claims 1-24, or the nanofiber of any one of claims 25-47, or the pharmaceutical composition of any one of claims 48-49.

51. A method of reducing inflammation in a subject, the method comprising administering to the subject a therapeutically effective amount of the allergen conjugate peptide of any one of claims 1-24, or the nanofiber of any one of claims 25-47, or the pharmaceutical composition of any one of claims 48-49.

52. The method of any one of claims 50-51, wherein the allergen conjugate peptide, the nanofiber, or the pharmaceutical composition as administered subcutaneously or sublingually or orally.Docket No.028193-0048-WO01 / 8355 53. The method of any one of claims 50-52, wherein the allergen does not leak into the vascular compartment or bloodstream.

54. The method of any one of claims 50-53, wherein the subject forms allergen-specific IgG antibodies.

55. The method of claim 54, wherein the allergen-specific IgG antibodies are detected in the subject for at least 20 weeks, at least 21 weeks, at least 22 weeks, at least 23 weeks, at least 24 weeks, at least 25 weeks, at least 30 weeks, at least 35 weeks, at least 40 weeks, at least 45 weeks, at least 50 weeks, at least 51 weeks, or at least 52 weeks.

56. The method of any one of claims 50-55, wherein the subject becomes desensitized to the allergen.

57. The method of any one of claims 50-56, wherein the subject has a reduced Type I hypersensitivity reaction to the allergen.

58. The method of any one of claims 50-57, wherein activation of bone marrow-derived mast cells (BMMC) in the subject is reduced.

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