Recombinant phages, probiotic vaccines and therapeutics, related compositions and methods of use

Abstract

Provided herein are recombinant phages, probiotic vaccines and therapeutics comprising the recombinant phages, and methods of use thereof.

Description

[Technical Field]

[0001] The present invention relates to recombinant phages, probiotic vaccines, and methods of their use.

[0002] Cancer is becoming more common today as a result of genetic abnormalities as well as epigenetic changes in cell development and apoptotic pathways.

[0003] Tumor development and growth is caused by the overexpression of genes associated with cancer cell growth and proliferation, and the underexpression of genes that control cell death activity.

[0004] Alterations in cellular pathways are frequently caused by random mutations or unhealthy lifestyles. Immunocompetent individuals are vulnerable to alterations in innate immunity, resulting in the activation of diverse immune responses and the destruction of normal tumor cells. Tumor-causing cells undergo apoptosis and necrosis, and surrounding tissues are also stressed, leading to the intestinal microbiota generating anti-cancer immune responses and signals.

[0005] Immunotherapy is a form of cancer treatment that activates the immune system to attack and eliminate cancer cells. Cytotoxic T lymphocytes (CTLs) are essential for a successful antitumor immune response. T cells that attack cancer cells must present tumor antigens to naive T cells, which undergo activation, clonal expansion, and ultimately exert cytotoxic effector function. Effective antigen presentation is essential for successful CTL effector function. Therefore, developing effective strategies to initiate tumor antigen presentation to T cells could be important for immunotherapy strategies in cancer treatment.

[0006] Similarly, B cells can inhibit tumor progression by promoting the production of tumor-reactive antibodies, tumor killing by NK cells, phagocytosis by macrophages, and priming of CD4+ and CD8+ T cells.B cells can promote tumor progression through the production of autoantibodies and tumor growth factors.

[0007] Live microbial cells that enter the gastrointestinal tract and confer beneficial effects on the host are called probiotics. Because the clinical outcomes of many types of cancer range from poor to fatal, there is a strong need to develop new preventative and / or therapeutic probiotic therapies. Summary of the Invention

[0008] The present invention provides a probiotic vaccine comprising a recombinant phage (e.g., a filamentous phage) and / or a recombinant filamentous phage genome, as well as a therapeutic method using the recombinant phage and / or probiotic vaccine of the present invention. The recombinant filamentous phage of the present invention and a probiotic vaccine comprising the recombinant phage are useful in therapeutic methods for preventing, treating, and / or reducing the risk of, for example, cancer, autoimmune diseases, respiratory diseases (e.g., asthma), neurological diseases, infectious diseases, and other diseases (osteoporosis, hypercholesterolemia, etc.).

[0009] Because SIgA antibodies are present in mucosal areas, while IgA and IgG antibodies are present in the circulation, immune responses within the circulatory system and mucosa are known to differ. Monomeric IgA is present in serum, whereas secretory IgA (SIgA) is present in mucosal secretions. SIgA generally exists in a polymeric form stabilized by linking chains (J chains), particularly as a dimer or tetramer setup, and therefore differs in structure from IgA present in serum. Furthermore, SIgA contains a secretory component (SC) derived from the polymeric Ig receptor (pIgR), which is utilized for crossing epithelial cells during secretion. In humans, there are two subclasses of IgA: IgA1 and IgA2. While the IgA1 subclass predominates in serum, the ratio of IgA1 to IgA2 in mucosal secretions varies depending on the site of production: for example, up to 60% of IgA1 is in saliva, up to 90% in nasal secretions, and up to 60% in intestinal secretions. In human colostrum, approximately 48% of immunoglobulins correspond to the IgA2 subclass and 40% to the IgA1 subclass, conferring adaptations to protect against potentially harmful pathogens as well as a means of controlling the colonization of the neonatal gut microbiota.

[0010] The mucous membranes lining the gastrointestinal, respiratory and urogenital tracts are in constant contact with a wide variety of microorganisms. The gastrointestinal tract (GIT) is home to a large and diverse microbial community, with up to approximately 10 14 The intestinal lining contains microbial cells, composed of approximately 500-1000 species of bacteria, archaea, and fungi. To prevent the invasion of pathogenic microorganisms and control host-bacterial interactions, the mucosal immune system is stimulated to produce SIgA. Mass production of SIgA in the intestine is one of the mechanisms by which the gut-associated lymphoid tissue (GALT) responds to the constant contact of the intestinal mucosa with a large number of diverse microorganisms (mainly bacteria and food-derived antigens).

[0011] In certain embodiments, probiotic vaccines of the invention comprising recombinant bacteriophages of the invention have been shown to have the ability to penetrate mucosal layers and generate significant secretory IgA (SIgA) immune responses, enabling the body to respond to intramucosal antigens and malignancies (see Figure 3 and Example 10). In another embodiment, recombinant bacteriophages of the invention (e.g., filamentous phage) have been shown to have the ability to generate significant IgA and IgG immune responses when injected and / or administered intravenously (IV) (see Figures 2A and 2B and Example 9).

[0012] Thus, provided herein are methods for preventing, treating, and / or reducing the risk of cancer, autoimmune diseases, respiratory diseases (e.g., asthma), neurological diseases, infectious diseases, and other diseases (e.g., osteoporosis, hypercholesterolemia, and the like), which comprise administering (e.g., oral administration, etc.) the probiotic vaccine of the present invention. Also provided herein are methods for preventing, treating, and / or reducing the risk of cancer, autoimmune diseases, respiratory diseases (e.g., asthma), neurological diseases, infectious diseases, and other diseases (e.g., osteoporosis, hypercholesterolemia, and the like), which comprise administering (e.g., injection, etc.) the recombinant bacteriophage of the present invention. In another aspect, provided herein are methods for preventing, treating, and / or reducing the risk of cancer, autoimmune diseases, respiratory diseases (e.g., asthma), neurological diseases, infectious diseases, and other diseases (particularly osteoporosis, hypercholesterolemia, and the like), which comprise administering (e.g., oral administration, etc.) the probiotic vaccine of the present invention in combination with the recombinant bacteriophage of the present invention (e.g., injection, etc.).

[0013] For example, in certain embodiments, the recombinant filamentous phage, probiotic vaccines, and therapeutic methods of the invention have shown significant effects on tumor growth in preclinical animal studies, and the therapeutic approaches of the invention are contemplated herein for addressing cancers endemic to the mucosa as well as slowing the metastasis of tumors arising elsewhere.

[0014] Accordingly, provided herein are methods for preventing, treating and / or reducing the risk of cancer, autoimmune diseases, respiratory diseases, neurological diseases and / or infectious diseases, comprising administering to a subject in need thereof: a. an injectable composition comprising a recombinant phage or probiotic vaccine comprising at least one exogenous peptide epitope corresponding to SEQ ID NOs: 9-133, or a fragment or variant thereof; and b. An oral composition comprising a recombinant phage or probiotic vaccine comprising at least one exogenous peptide epitope corresponding to SEQ ID NOs: 9-133, or a fragment or variant thereof. The method comprises administering a combination of: Also provided are methods for preventing, treating, and / or reducing the risk of cancer, autoimmune diseases, respiratory diseases, neurological diseases, and / or infectious diseases, comprising administering to a subject in need thereof a probiotic vaccine of the present invention described herein. In certain embodiments, the probiotic vaccine is administered orally. In another embodiment, the method of the present invention further comprises administering a recombinant bacteriophage described herein. In certain embodiments, the recombinant phage is administered intravenously or intramuscularly. In yet another embodiment, secretory IgA (SIgA) is produced. In still another embodiment, secretory IgA (SigA) is produced in an amount 2-100-fold, 2-90-fold, 2-80-fold, 2-70-fold, 2-60-fold, 2-50-fold, 2-40-fold, 2-30-fold, 2-20-fold, 2-10-fold, 2-9-fold, 2-8-fold, 2-7-fold, 2-6-fold, 2-5-fold, or 2-4-fold greater than the control.

[0015] Also provided herein is a probiotic vaccine comprising a recombinant filamentous phage, said phage comprising: ALLEIASCL (SEQ ID NO: 9)-IDO1, MYPPPY (SEQ ID NO: 10)-CTLA-4, NPLVA (SEQ ID NO: 11) - claudin, ESKVDPSK (SEQ ID NO: 12) - prostate-specific membrane antigen (PSMA) mimitope, RSSSRTPS (SEQ ID NO: 13) - TNF epitope, AGICAS (SEQ ID NO: 14) - COVID-19 epitope, AG-I-CAS (SEQ ID NO: 15; Ile = "AUA" codon) AKPTAKREAIVKAYG (SEQ ID NO: 16) - monkeypoxvirus, ISPDGCYSL (SEQ ID NO: 17) - monkeypoxvirus, LTFDYVVTF (SEQ ID NO: 18) - monkeypoxvirus, VLYQDVNCT (SEQ ID NO: 19) - Covid Wuhan Spike, QQQQGQTITKKSAAEASKKPRQKRTAVLVQDVGCTFSTVFPVTSFGPLVR (SEQ ID NO: 20) - CovidItaly org9, VLYQGVNCT (SEQ ID NO: 21) - Covid Italy spike, DAPF (SEQ ID NO: 22) - avian influenza, KQIVERILEEE (SEQ ID NO: 23) - avian influenza, RYLTDMTLEE (SEQ ID NO: 24) - avian influenza, DDVKRENPEHEEYNN (SEQ ID NO: 25) - avian influenza, RESGSS (SEQ ID NO: 26) - avian influenza, FRHDSGY (SEQ ID NO: 27) - amyloid-beta AGICASAGICAS (SEQ ID NO: 28) - COVID-19 vaccine ESKVDPSKAW (SEQ ID NO: 29) - prostate-specific membrane antigen (PSMA), DAPFKQIVERILEEE (SEQ ID NO: 30) - avian influenza, RESGSSRYLTDMTLEE (SEQ ID NO: 31) - avian influenza, DDVKRENPEHEEYNN (SEQ ID NO: 32) - avian influenza, CVADYSVLY (SEQ ID NO: 33) - SARS-CoV2, HVGEIPVAY (SEQ ID NO: 34) - SARS-CoV2, NQKLIANQF(NQK-Q8) (SEQ ID NO: 35)-NQK-Q8, SARS-CoV2; NQKLIANAF(NQK-A8) (SEQ ID NO: 36)-NQK-A8, SARS-CoV2, RVAGDSGFAAY (SEQ ID NO: 37) - SARS-CoV2, ESFDGDPASNTAPLQPEQL (SEQ ID NO: 38)-PD-1, EFINVPEGGS (SEQ ID NO: 39) - avian influenza, EFINVPEEFINVPE (SEQ ID NO: 40) - avian influenza, VRSSSRTPSD (SEQ ID NO: 41) - TNF-α inhibitor, RSSSRTPSDK (SEQ ID NO: 42) - TNF-α inhibitor, MYPPPYMYPPPY (SEQ ID NO: 43)-CTLA-4, VELMYPPPYYLGIG (SEQ ID NO: 44)-CTLA-4, LMYPPPYYLGIGN (SEQ ID NO: 45)-CTLA-4, NPLVANPLVA (SEQ ID NO: 46) - Claudin (epithelial ovarian cancer), LPNAIGRGKWWRP (SEQ ID NO: 47) - sclerostin epitope, ETPEGAEA (SEQ ID NO: 48)-TNF-α, KRNIQFSCKNI (SEQ ID NO: 49)-CD38, LTFDY (SEQ ID NO: 50) - monkeypoxvirus, LTFDYLTFDY (SEQ ID NO: 51) - monkeypoxvirus, NTVSCNRPHCITE (SEQ ID NO: 52) - asthma, EERRR (SEQ ID NO: 53) - asthma, EERRREERRR (SEQ ID NO: 54) - asthma, LTHVKIND (SEQ ID NO: 55) - yellow fever, NANPNANPNANPNAN (SEQ ID NO: 56) - malaria, EIRQAGRPNKPDSI (SEQ ID NO: 57)-TL1A, QTLEAQTLEA (SEQ ID NO: 58) - CD38, IQFSCIQFSC (SEQ ID NO: 59) - CD38, DAPFDAPF (SEQ ID NO: 60) - avian influenza, RESGSSRESGSS (SEQ ID NO: 61) - avian influenza, VVVGADVGKS (SEQ ID NO: 62) - KRAS, PTPEEDQILKK (SEQ ID NO: 63)-BTKi, APKAQIKESLR (SEQ ID NO: 64)-PD1, ASPLLGSQHFDVPPRQL (SEQ ID NO: 65)-XPO1, QYDSWPHYPE (SEQ ID NO: 66)-LOXL2, KVIQAQTAFSANPANPAILS (SEQ ID NO: 67) - Syntenin MDA9, PIPHDGNLYPRLYPE (SEQ ID NO: 68) - Syntenin MDA9, SKCRRCRLLD (SEQ ID NO: 69)-CD95, QNTKCRCKPNFFC (SEQ ID NO: 70)-CD95, LCTLAWAHPLVGHM (SEQ ID NO: 71)-IL-23, ESVDPKNYPKK (SEQ ID NO: 72)-IL-1b, YLSETVSNALGPQGRRLEE (SEQ ID NO: 73)-LOXL4, INDPRLRNPSDKFIY (SEQ ID NO: 74)-NMDA1, SPRMSGLLSQTGYLSDNHILI (SEQ ID NO: 75)-DLL3, GLPSLGLTL (SEQ ID NO: 76) - nerve growth receptor, HRGEFS (SEQ ID NO: 77) - nerve growth factor, HRGEFSHRGEFS (SEQ ID NO: 78) - nerve growth factor, THLHWHTV (SEQ ID NO: 79)-β-amyloid, VQDYVQMKASELEQ (SEQ ID NO: 80) - calcitonin, REIEGRVMVTDPENVPEEDG (SEQ ID NO: 81)-PCSK9 (LDL cholesterol), AGAFAPSAAVA (SEQ ID NO: 82)-IL-4, GYYYRDDSLPVLLE (SEQ ID NO: 83) - retinoic acid (acne), EDPAFYMLKGKLQYELQ (SEQ ID NO: 84)-IL-21, ALFDAQAQV (SEQ ID NO: 85)-TYK2, LLYDLTEKQVY (SEQ ID NO: 86)-TYK2, ALDENTKLWYA (SEQ ID NO: 87)-JAK1, VLVESEHQV (SEQ ID NO: 88)-JAK1, APNRTITVDDKMSLRLH (SEQ ID NO: 89)-JAK1, SSDWVTLGTSASLLR (SEQ ID NO: 90) - Ceacam 1, STPYVGLSLSCAANQ (SEQ ID NO: 91) - Ceacam 1, PSPQYSWRINGIPQQHTQVLF (SEQ ID NO: 92) - Ceacam 5, STLYREELMTTLLYYEIFL (SEQ ID NO: 93)-TRPV1, CEKNSVLEVIAYSSSE (SEQ ID NO: 94) - TRPV1 Sub V, NPTTYQMDVNPEGKYS (SEQ ID NO: 95)-EGFR, HTPPLDPQELDILKT (SEQ ID NO: 96) - EGFR, RPKSNIVLLS (SEQ ID NO: 97)-CD-20, SGPKPLFRRMSSLVGPTQ (SEQ ID NO: 98)-CD-20, RPKSNIVLLSAEEKKEQT (SEQ ID NO: 99)-CD-20, DYEDRYYRE (SEQ ID NO: 100) - prion, RPLKGAPLPAL (SEQ ID NO: 101)-ALK, MTEQQWNFAGIEAAASAIQG (SEQ ID NO: 102) - tuberculosis, KTIAYDEEARR (SEQ ID NO: 103) - leprosy, EEFVVEFDLPGIKA (SEQ ID NO: 104) - leprosy, CVTIMTVDNPASTTNKDKLFAVWKITYKDT (SEQ ID NO: 105) - poliovirus 1, ILKEPVHGV (SEQ ID NO: 106)-HIV1, SLYNTVATL (SEQ ID NO: 107) - HIV1, LSEIKGVIVHRLEGV (SEQ ID NO: 108) - measles, MLMAASRAL (SEQ ID NO: 109) - Yersinia pestis, RAMRMVYYL (SEQ ID NO: 110) - Yersinia pestis, FFVRSIGSKGGKLAAGKYTDAVTV (SEQ ID NO: 111) - Yersinia pestis, EEPEESDEDDFGMGGLF (SEQ ID NO: 112) - Leishmania chagasi, CINGVCWTV (SEQ ID NO: 113) - Hepatitis C, KLVALGINAV (SEQ ID NO: 114) - Hepatitis C, NLVPMVATV (SEQ ID NO: 115) - shingles, TPRVTGGGAM (SEQ ID NO: 116) - shingles, TETEPPVAL (SEQ ID NO: 117)-ESRRA, REALHEAL (SEQ ID NO: 118)-ESRRA, DVIGRIENGERL (SEQ ID NO: 119)-TKI, leukemia epitope 2, RLIKEDVYL (SEQ ID NO: 120)-TKI, leukemia epitope 6, KTLETNPYVK (SEQ ID NO: 121)-Ku70, GRADALRVLSK (SEQ ID NO: 122)-IGF-1R, LSRELSSLASQPLSR (SEQ ID NO: 123)-IGF-1R, SLLPDSPVGQ (SEQ ID NO: 124)-IL-23, SPSQPWQRL (SEQ ID NO: 125)-IL-23, FLAEDALNTV (SEQ ID NO: 126) - breast cancer, CTHYMQLAC (SEQ ID NO: 127) - osteoporosis, QEANASAQTKCDIE (SEQ ID NO: 128) - Alaskapoxvirus, VVVGACVGKS (SEQ ID NO: 129) - KRAS, LPAIVREAI (SEQ ID NO: 130) - Dengue virus, SPRRKLISVDSRSVSL (SEQ ID NO: 131)-IL-21, DKPVYTPDQSVKVRV (SEQ ID NO: 132)-C5, and / or DNVLDHLTGRSC (SEQ ID NO: 133) - pertussis; and Bacteria infected with the recombinant phage Also provided is said probiotic vaccine, comprising a nucleic acid encoding a recombinant polypeptide comprising an exogenous peptide epitope selected from the group consisting of:

[0016] In certain embodiments of the vaccines of the present invention, the exogenous peptide epitope is functionally expressed on a coat protein selected from the group consisting of pIII, pVI, pVII, pVIII, and pIX. In certain embodiments, the coat protein is pIII. In other embodiments, the coat protein is pVIII.

[0017] In further embodiments, the phage is selected from the group consisting of M13, fd, IKe, CTX-φ, Pf1, Pf2, Pf3, f1, MKE, M13KE, type 8, type f88, f88-4 (GenBank Accession # AF218363), Myoviridae (P1-like viruses, P2-like viruses, Mu-like viruses, SPO1-like viruses, phiH-like viruses), Siphoviridae (λ-like viruses, γ-like viruses, T1-like viruses, T5-like viruses, c2-like viruses, L5-like viruses, psiM1-like viruses, phiC31-like viruses, N15-like viruses), Podoviridae (phi29-like viruses, P22-like viruses, N4-like viruses), Tectiviridae (tectiviruses), Corticoviridae (corticoviruses), Liposthrixviridae (alphalipothrixviruses, betalipothrixviruses), The phage is selected from the group consisting of filamentous phages, including Liposthrixvirus, Gammaliposthrixvirus, Deltaliposthrixvirus, Plasmaviridae (plasma viruses), Rudiviridae (rudivruria), Fuselloviridae (fuselloviruses), Inoviridae (inoviruses, plectroviruses, M13-like viruses, fd-like viruses), Microviridae (microviruses, spiromicroviruses, bdelomicroviruses, chlamydiamicroviruses), Leviviridae (leviviruses, alloleviviruses), and Cystoviridae (cystoviruses). In a specific embodiment, the phage is M13KE. In another embodiment, the phage is f88 or f88-4.

[0018] In yet another embodiment, the bacteria (e.g., probiotic bacteria) to be infected with the recombinant phage of the invention provided herein are selected from the group consisting of E. coli Nissle 1917, E. coli ER2738, Bacillus amyloliquefaciens, Bacillus polyfermenticus Bispan, Bifidobacterium animalis subsp. lactis BB-12, Bifidobacterium animalis subsp. lactis GPS1209, Bifidobacterium animalis subsp. lactis HN019 (DR1064) strain, Bifidobacterium bifidum strain BB-12, Bifidobacterium bifidum strain Rosell-71, Bifidobacterium breve strain M-16V, Bifidobacterium longum, Bifidobacterium thermophilum, Lactobacillus acidophilus strain La-1, Lactobacillus brevis strain HA-112, Lactobacillus fermentum HA-179 strain, Lactobacillus helveticus Lafti strain L10, Lactobacillus helveticus Rosell-52 strain, Lactobacillus paracasei Lafti strain L26, Lactobacillus paracasei subsp.Lactobacillus paracasei subsp. paracasei strain 431, Lactobacillus rhamnosus strain HN001(DR20), Streptococcus salivarius strain DSM 13084, Streptococcus thermophilus, Bacillus coagulans strain GBI-30,6086, Bifidobacterium animalis subsp. lactis strain BB-12, Bifidobacterium longum subsp. infantis, Escherichia coli Nissle 1917, Escherichia coli Lactobacillus acidophilus strain ER2738, Lactobacillus acidophilus strain NCFM, Lactobacillus paracasei strain Stl 1 (or strain NCC2461), Lactobacillus johnsonii strain Lai (also called Lactobacillus strain LCI or Lactobacillus johnsonii strain NCC533), Lactobacillus plantarum strain 299v, Lactobacillus reuteri strain ATCC 55730 (Lactobacillus reuteri strain SD2112), Lactobacillus reuteri strain Protectis (DSM 17938, a daughter strain of ATCC 55730), Lactobacillus reuteri Prodentis (a combination of DSM 17938 / ATCC 55730 and ATCC PTA5289), Lactobacillus rhamnosusGG strain, Saccharomyces boulardii, a mixture of Lactobacillus rhamnosus strain GR-1 and Lactobacillus reuteri strain RC-14, a mixture of Lactobacillus acidophilus strain NCFM and Bifidobacterium bifidum strain BB-12, a mixture of Lactobacillus acidophilus strain CL1285 and Lactobacillus casei strain LBC80R, and a mixture of Lactobacillus plantarum strain HEAL 9 and Lactobacillus paracasei. 8700:2 strain, a mixture of Lactobacillus bulgaricus, Lactococcus thermophiles and Lactobacillus bifidus.

[0019] In a specific embodiment, the probiotic vaccine bacterium infected with the recombinant phage of the present invention described herein is F factor positive. In another embodiment, the bacterium is an F factor positive strain selected from Escherichia coli Nissle 1917 or Escherichia coli ER2738. In a specific embodiment, the bacterium is Escherichia coli Nissle 1917. In another embodiment, the bacterium is Escherichia coli ER2738. Escherichia coli Nissle 1917 (EcN) is a non-pathogenic strain of Enterobacteriaceae (Creative Biolabs).

[0020] In certain embodiments, the probiotic vaccine and / or recombinant phage produces both IgG and secretory IgA (SIgA) antibodies that bind to the exogenous peptide epitope.

[0021] Also provided herein is a recombinant filamentous phage comprising a recombinant phage genome comprising a nucleic acid encoding a polypeptide comprising an exogenous peptide epitope selected from the group consisting of SEQ ID NOs: 9 to 133, or a fragment or variant thereof.

[0022] In certain embodiments of the recombinant phage, the exogenous peptide epitope is functionally expressed on a coat protein selected from the group consisting of pIII, pVI, pVII, pVIII, and pIX. In certain embodiments of the recombinant phage, the coat protein is pIII. In another embodiment, the coat protein is pVIII. In further embodiments, the phage is selected from the group consisting of M13, fd, IKe, CTX-φ, Pf1, Pf2, Pf3, f1, MKE, M13KE, type 8, type f88, f88-4, Myoviridae (P1-like viruses, P2-like viruses, Mu-like viruses, SPO1-like viruses, phiH-like viruses), Siphoviridae (λ-like viruses, γ-like viruses, T1-like viruses, T5-like viruses, c2-like viruses, L5-like viruses, psiM1-like viruses, phiC31-like viruses, N15-like viruses), Podoviridae (phi29-like viruses, P22-like viruses, N4-like viruses), Tectiviridae (tectiviruses), Corticoviridae (corticoviruses), In yet another embodiment, the phage is selected from the group consisting of filamentous phage, including Liposthrixviridae (Alphalipothrixvirus, Betalipothrixvirus, Gammalipothrixvirus, Deltalipothrixvirus), Plasmaviridae (Plasmaviruses), Rudiviridae (Rudivruridae), Fuselloviridae (Fucelloviruses), Inoviridae (Inoviruses, Plectroviruses, M13-like viruses, fd-like viruses), Microviridae (Microviruses, Spiromicroviruses, Bdelomicroviruses, Chlamydiamicroviruses), Leviviridae (Leviviruses, Alloleviviruses), and Cystoviridae (Cystoviruses). In yet another embodiment, the phage is a filamentous phage selected from the group consisting of M13, fd, IKe, CTX-φ, Pf1, Pf2, Pf3, f1, MKE, and M13KE. In a specific embodiment, the phage is M13KE. In another embodiment, the phage is f88 or f88-4. In yet another embodiment, the recombinant phage produces IgG antibodies that bind to the exogenous peptide epitope.

[0023] Also provided herein are methods of preventing, treating, and / or reducing the risk of cancer, comprising administering to a subject in need thereof one or more of the probiotic vaccines described herein or one or more of the recombinant phages described herein. In certain embodiments, the cancer is selected from the group consisting of multiple myeloma, epithelial cancer, epithelial ovarian cancer, mucosal melanoma, non-small cell lung cancer, melanoma, head and neck cancer, renal cell carcinoma, Hodgkin's lymphoma, cutaneous squamous cell carcinoma, glioblastoma, esophageal cancer, gastric cancer, duodenal cancer, small intestine cancer, appendix cancer, colon cancer, rectal cancer, colorectal cancer, anal cancer, pancreatic cancer, liver cancer, gallbladder cancer, spleen cancer, kidney cancer, bladder cancer, prostate cancer, testicular cancer, uterine cancer, endometrial cancer, ovarian cancer, vaginal cancer, vulvar cancer, breast cancer, lung cancer, thyroid cancer, thymus cancer, brain cancer, and thyroid cancer. The cancer is selected from the group consisting of: systemic cancer, glioma, oral cancer, skin cancer, blood cancer, lymphoma, eye cancer, bone cancer, bone marrow cancer, muscle cancer, non-small cell lung cancer (NSCLC), head and neck squamous cell carcinoma (HNSCC), urothelial cancer, non-muscle-invasive bladder cancer (NMIBC), colon or rectal cancer, esophageal cancer or certain gastroesophageal junction (GEJ) cancers, cervical cancer, renal cell carcinoma (RCC), advanced endometrial cancer, cutaneous squamous cell carcinoma (cSCC), and / or triple-negative breast cancer (TNBC), etc. In certain embodiments, the cancer is multiple myeloma.

[0024] Also provided herein are further methods as set forth below in items 23-60: 23. A method for preventing or treating cancer, comprising administering to a subject in need thereof one or more of the probiotic vaccines of the invention described herein or one or more of the recombinant phages of the invention described herein, wherein the exogenous peptide epitope is selected from the group consisting of SEQ ID NOs: 9-12, 29, 38, 43-46, 49, 58, 59, 62-70, 73, 75, 84, 90-92, 95, 96, 101, 117-123, 126, 129 and / or 131. 24. The cancer is multiple myeloma, epithelial cancer, epithelial ovarian cancer, mucosal melanoma, non-small cell lung cancer, melanoma, head and neck cancer, renal cell carcinoma, Hodgkin's lymphoma, cutaneous squamous cell carcinoma, glioblastoma, esophageal cancer, gastric cancer, duodenal cancer, small intestine cancer, appendix cancer, colon cancer, rectal cancer, colorectal cancer, anal cancer, pancreatic cancer, liver cancer, gallbladder cancer, spleen cancer, kidney cancer, bladder cancer, prostate cancer, testicular cancer, uterine cancer, endometrial cancer, ovarian cancer, vaginal cancer, vulvar cancer, breast cancer, lung cancer, thyroid cancer, thymus cancer, brain cancer, nervous system cancer, or glioma. , oral cancer, skin cancer, blood cancer, lymphoma, eye cancer, bone cancer, bone marrow cancer, muscle cancer, non-small cell lung cancer (NSCLC), head and neck squamous cell carcinoma (HNSCC), urothelial cancer, non-muscle invasive bladder cancer (NMIBC), colon or rectal cancer, esophageal cancer or certain gastroesophageal junction (GEJ) cancer, cervical cancer, renal cell carcinoma (RCC), advanced endometrial cancer, cutaneous squamous cell carcinoma (cSCC) and / or triple-negative breast cancer (TNBC). 25. The method of item 23 or 24, wherein the cancer is multiple myeloma. 26. A method for preventing, treating and / or reducing the risk of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer and / or gastric cancer, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from NPLVA (SEQ ID NO: 11), NPLVANPLVA (SEQ ID NO: 46) and / or FLAEDALNTV (SEQ ID NO: 126), or a fragment or variant thereof. 27. The method according to item 26, wherein a recombinant phage and / or probiotic vaccine comprising FLAEDALNTV (SEQ ID NO: 126) is used to prevent, treat or reduce the risk of breast cancer. 28. 1. A method of preventing, treating and / or reducing the risk of multiple myeloma, comprising administering to a subject in need thereof: KRNIQFSCKNI (SEQ ID NO: 49), QTLEAQTLEA (SEQ ID NO: 58), and / or IQFSCIQFSC (SEQ ID NO: 59) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of: 29. A method for preventing, treating and / or reducing the risk of cancer, comprising administering to a subject in need thereof a recombinant phage comprising at least one exogenous peptide epitope selected from the group consisting of ESFDGDPASNTAPLQPEQL (SEQ ID NO: 38), APKAQIKESLR (SEQ ID NO: 64), EDPAFYMLKGKLQYELQ (SEQ ID NO: 84) and / or SPRKLISVDSRSVSL (SEQ ID NO: 131), or a fragment or variant thereof. and / or a probiotic vaccine, wherein the cancer is selected from the group consisting of mucosal cancer, gastric cancer, melanoma, non-small cell lung cancer (NSCLC), head and neck squamous cell carcinoma (HNSCC), urothelial carcinoma, non-muscle-invasive bladder cancer (NMIBC), colon or rectal cancer, esophageal cancer or certain gastroesophageal junction (GEJ) cancers, cervical cancer, renal cell carcinoma (RCC), advanced endometrial cancer, cutaneous squamous cell carcinoma (cSCC), and / or triple-negative breast cancer (TNBC). 30. A method of preventing, treating, and / or reducing the risk of mucosal melanoma, comprising administering to a subject in need thereof: ALLEIASCL (SEQ ID NO: 9), EDPAFYMLKGKLQYELQ (SEQ ID NO: 84), MYPPPY (SEQ ID NO: 10), MYPPPYMYPPPY (SEQ ID NO: 43), VELMYPPPYYLGIG (SEQ ID NO: 44), LMYPPPYYLGIGN (SEQ ID NO: 45), and / or SPRRKLISVDSRSVSL (SEQ ID NO: 131) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of: 31. A method of preventing, treating, and / or reducing the risk of prostate cancer, comprising administering to a subject in need thereof: ESKVDPSKAW (SEQ ID NO: 29), ESKVDPSK (SEQ ID NO: 12), MYPPPY (SEQ ID NO: 10), MYPPPYMYPPPY (SEQ ID NO: 43), VELMYPPPYYLGIG (SEQ ID NO: 44), LMYPPPYYLGIGN (SEQ ID NO: 45), ASPLLSGSQHFDVPPRQL (SEQ ID NO: 65), and / or KVIQAQTAFSANPANPAILS (SEQ ID NO: 67) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of: 32. A method for preventing, treating, and / or reducing the risk of blood cancer, leukemia, chronic lymphocytic leukemia (CLL), small lymphocytic lymphoma (SLL), Waldenstrom's macroglobulinemia, chronic graft-versus-host disease, mantle cell lymphoma (MCL), marginal zone lymphoma, etc., in a subject in need thereof, comprising: PTPEEDQILKK (SEQ ID NO: 63), DVIGRIENGERL (SEQ ID NO: 119), and / or RLIKEDVYL (SEQ ID NO: 120) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of: 33. A method for preventing, treating and / or reducing the risk of acute myeloid leukemia (AML), autoimmune and inflammatory diseases, canine cancer, etc., comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope corresponding to ASPLLSGSQHFDVPPRQL (SEQ ID NO: 65), or a fragment or variant thereof. 34. A method of preventing, treating, and / or reducing the risk of pulmonary fibrosis in a subject in need thereof: QYDSWPHYPE (SEQ ID NO: 66), YLSETVSNALGPQGRRLEE (SEQ ID NO: 73), EDPAFYMLKGKLQYELQ (SEQ ID NO: 84), MYPPPY (SEQ ID NO: 10), MYPPPYMYPPPY (SEQ ID NO: 43), VELMYPPPYYLGIG (SEQ ID NO: 44), LMYPPPYYLGIGN (SEQ ID NO: 45), and / or SPRRKLISVDSRSVSL (SEQ ID NO: 131) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of: 35. A method for preventing, treating, and / or reducing the risk of lung cancer, non-small cell lung cancer, etc., comprising administering to a subject in need thereof: SPRMSGLLSQTGYLSDNHILI (SEQ ID NO: 75), ASPLLSGSQHFDVPPRQL (SEQ ID NO: 65), SSDWVTLGTSASLLR (SEQ ID NO: 90), STPYVGLSLSCAANQ (SEQ ID NO: 91), PSPQYSWRINGIPQQHTQVLF (SEQ ID NO: 92), NPTTYQMDVNPEGKYS (SEQ ID NO: 95), HTPPLDPQELDILKT (SEQ ID NO: 96), RPLKGAPLPAL (SEQ ID NO: 101), EDPAFYMLKGKLQYELQ (SEQ ID NO: 84), MYPPPY (SEQ ID NO: 10), VVVGADVGKS (SEQ ID NO: 62), VVVGACVGKS (SEQ ID NO: 129), MYPPPYMYPPPY (SEQ ID NO: 43), VELMYPPPYYLGIG (SEQ ID NO: 44), LMYPPPYYLGIGN (SEQ ID NO: 45), GRADALRVLSK (SEQ ID NO: 122), LSRELSSLASQPLSR (SEQ ID NO: 123), and / or SPRRKLISVDSRSVSL (SEQ ID NO: 131) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of: 36. A method for preventing, treating, and / or reducing the risk of thyroid eye disease, etc., comprising administering to a subject in need thereof: GRADALRVLSK (SEQ ID NO: 122), and / or LSRELSSLASQPLSR (SEQ ID NO: 123) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of: 37. A method of preventing, treating, and / or reducing the risk of cancer, comprising administering to a subject in need thereof: PIPHDGNLYPRLYPE (SEQ ID NO: 68), SKCRRCRLLD (SEQ ID NO: 69), QNTKCRCKPNFFC (SEQ ID NO: 70), TETEPPVAL (SEQ ID NO: 117), REALHEAL (SEQ ID NO: 118), VVVGADVGKS (SEQ ID NO: 62), VVVGACVGKS (SEQ ID NO: 129), and / or KTLETNPYVK (SEQ ID NO: 121) administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of: The method, wherein the cancer is selected from the group consisting of gastric cancer, melanoma, mucosal melanoma, lung cancer, non-small cell lung cancer (NSCLC), head and neck squamous cell carcinoma (HNSCC), urothelial carcinoma, non-muscle-invasive bladder cancer (NMIBC), colon or rectal cancer, esophageal cancer or certain gastroesophageal junction (GEJ) cancers, cervical cancer, renal cell carcinoma (RCC), advanced endometrial cancer, cutaneous squamous cell carcinoma (cSCC), pancreatic ductal carcinoma, and / or prostate cancer. 38. The method of item 37, wherein the recombinant phage and / or probiotic vaccine comprises KTLETNPYVK (SEQ ID NO: 121) and is used to prevent, treat or reduce the risk of colorectal cancer. 39. A method for preventing Korean fever virus infection and / or disease manifestations, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of: KRNIQFSCKNI (SEQ ID NO: 49), QTLEAQTLEA (SEQ ID NO: 58) and / or IQFSCIQFSC (SEQ ID NO: 59), or a fragment or variant thereof. 40. A method of preventing COVID-19 infection and / or disease manifestations in a subject in need thereof, comprising administering: AGICAS (SEQ ID NO: 14), AG-I-CAS (SEQ ID NO: 15; Ile = "AUA" codon), VLYQDVNCT (SEQ ID NO: 19), QQQQGQTITKKSAAEASKKPRQKRTAVLVQDVGCTFSTVFPVTSFGPLVR (SEQ ID NO: 20), VLYQGVNCT (SEQ ID NO: 21), AGICASA GICAS (SEQ ID NO: 28), CVADYSVLY (SEQ ID NO: 33), HVGEIPVAY (SEQ ID NO: 34), NQKLIANQF (NQK-Q8) (SEQ ID NO: 35), NQKLIANAF (NQK-A8) (SEQ ID NO: 36), and / or RVAGDSGFAAY (SEQ ID NO: 37) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of: 41. A method for preventing, treating and / or reducing the risk of hepatitis C, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of CINGVCWTV (SEQ ID NO: 113) and / or KLVALGINAV (SEQ ID NO: 114), or a fragment or variant thereof. 42. A method for preventing, treating and / or reducing the risk of shingles, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of: NLVPMVATV (SEQ ID NO: 115) and / or TPRVTGGGAM (SEQ ID NO: 116), or a fragment or variant thereof. 43. A method for preventing, treating, and / or reducing the risk of avian influenza infection and / or disease manifestations, comprising administering to a subject in need thereof: DAPF (SEQ ID NO: 22), KQIVERILEEE (SEQ ID NO: 23), RYLTDMTLEE (SEQ ID NO: 24), DDVKRENPEHEEYNN (SEQ ID NO: 25), RESGSS (SEQ ID NO: 26), DAPFKQIVERILEEE (SEQ ID NO: 30), RESGSSRYLTDMTLEE (SEQ ID NO: 31), EFINVPEGGS (SEQ ID NO: 39), EFINVPEEFINVPE (SEQ ID NO: 40), DAPFDAPF (SEQ ID NO: 60), and / or RESGSSRESGSS (SEQ ID NO: 61) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of: 44. A method for preventing, treating, and / or reducing the risk of rheumatoid arthritis, juvenile idiopathic arthritis, psoriatic arthritis, ankylosing spondylitis, Crohn's disease, ulcerative colitis, plaque psoriasis, hidradenitis suppurativa, or uveitis in a subject in need thereof: RSSSRTPS (SEQ ID NO: 13), VRSSSRTPSD (SEQ ID NO: 41), RSSSRTPSDK (SEQ ID NO: 42), ETPEGAEA (SEQ ID NO: 48), and / or ALDENTKLWYA (SEQ ID NO: 87) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of: 45. A method of preventing, treating, and / or reducing the risk of multiple sclerosis in a subject in need thereof: ESVDPKNYPKK (SEQ ID NO: 72), RPKSNIVLLS (SEQ ID NO: 97), RPKSNIVLLSAEEKKEQT (SEQ ID NO: 99), and / or SGPKPLFRRMSSLVGPTQ (SEQ ID NO: 98) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of: 46. ​​A method for preventing, treating and / or reducing the risk of osteoporosis, sclerosis, Van Buchem disease, osteonecrosis, osteoarthritis, rheumatoid arthritis, ankylosing spondylitis, bone tumors, multiple myeloma, bone metastasis, prostate cancer, glucocorticosteroid-induced osteoporosis, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of LPNAIGRGKWWRP (SEQ ID NO: 47) and / or CTHYMQLAC (SEQ ID NO: 127), or a fragment or variant thereof. 47. A method for preventing, treating and / or reducing the risk of primary hyperlipidemia, heterozygous familial hypercholesterolemia, hyperlipidemia, myocardial infarction, stroke and coronary revascularization, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope corresponding to REIEGRVMVTDPENVPEEDG (SEQ ID NO: 81), or a fragment or variant thereof. 48. A method for preventing, treating and / or reducing the risk of acne, signs of aging, wrinkles, fine lines, scars, hyperpigmentation, Kaposi's sarcoma lesions, melasma, psoriasis, stretch marks, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope corresponding to GYYYRDDSLPVLLE (SEQ ID NO: 83), or a fragment or variant thereof. 49. A method for preventing, treating and / or reducing the risk of paroxysmal nocturnal hemoglobinuria, atypical hemolytic uremic syndrome, intravascular hemolysis, blood clots, fatigue and the need for blood transfusions, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope corresponding to DKPVYTPDQSVKVRV (SEQ ID NO: 132) or a fragment or variant thereof. 50. A method for preventing, treating and / or reducing the risk of asthma, severe asthma, atopic dermatitis, eczema, eosinophilic or oral steroid-dependent asthma, chronic rhinosinusitis with nasal polyps, eosinophilic esophagitis, prurigo nodularis, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of NTVSCNRPHCITE (SEQ ID NO: 52), EERRR (SEQ ID NO: 53), EERRREERRR (SEQ ID NO: 54) and / or AGAFAPSAAVA (SEQ ID NO: 82), or a fragment or variant thereof. 51. A method for preventing, treating and / or reducing the risk of asthma or severe eosinophilic asthma, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of (SEQ ID NO: 52), EERRR (SEQ ID NO: 53) and / or EERRREERRR (SEQ ID NO: 54), or a fragment or variant thereof. 52. A method of preventing, treating, and / or reducing the risk of psoriasis in a subject in need thereof: LCTLAWAHPLVGHM (SEQ ID NO: 71), ALFDAQAQV (SEQ ID NO: 85), LLYDLTEKQVY (SEQ ID NO: 86), APNRTITVDDKMSLRLH (SEQ ID NO: 89), SLLPDSPVGQ (SEQ ID NO: 124), and / or SPSQPWQRL (SEQ ID NO: 125) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of: 53. A method for preventing, treating, and / or reducing the risk of an inflammatory disease selected from the group consisting of psoriatic arthritis, Crohn's disease, ulcerative colitis, psoriasis, rheumatoid arthritis, atopic dermatitis, ankylosing spondylitis, hidradenitis suppurativa, and uveitis, comprising administering to a subject in need thereof: EIRQAGRPNKPDSI (SEQ ID NO: 57), and / or VLVESEHQV (SEQ ID NO: 88) The method comprises administering a recombinant phage or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of: 54. A method of preventing, treating, and / or reducing the risk of pain in a subject in need thereof, comprising: GLPSLGLTL (SEQ ID NO: 76), HRGEFS (SEQ ID NO: 77), HRGEFSHRGEFS (SEQ ID NO: 78), STLYREELMTTLLYYEIFL (SEQ ID NO: 93), and / or CEKNSVLEVIAYSSSE (SEQ ID NO: 94) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of: 55. A method for preventing, treating and / or reducing the risk of chronic wasting, mad cow disease, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope corresponding to DYEDRYYRE (SEQ ID NO: 100), or a fragment or variant thereof. 56. A method for preventing, treating, and / or reducing the risk of a neurological disorder selected from the group consisting of Alzheimer's disease, Parkinson's disease, dementia, multiple sclerosis, epilepsy, Huntington's disease, muscular dystrophy, and migraine, comprising administering to a subject in need thereof: FRHDSGY (SEQ ID NO: 27), THLHWHTV (SEQ ID NO: 79), VQDYVQMKASELEQ (SEQ ID NO: 80), and / or INDPRLRNPSDKFIY (SEQ ID NO: 74) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of: 57. The method according to item 58, wherein the disease is Alzheimer's disease. 58. The method according to item 56, wherein the disease is migraine and the recombinant phage and / or probiotic vaccine comprises at least one exogenous peptide epitope selected from VQDYVQMKASELEQ (SEQ ID NO: 80). 59. The method according to any one of items 23 to 58, wherein secretory IgA (SigA) is produced. 60. The method according to item 59, wherein secretory IgA (SigA) is produced in an amount that is 2 to 100 times, 2 to 90 times, 2 to 80 times, 2 to 70 times, 2 to 60 times, 2 to 50 times, 2 to 40 times, 2 to 30 times, 2 to 20 times, 2 to 10 times, 2 to 9 times, 2 to 8 times, 2 to 7 times, 2 to 6 times, 2 to 5 times, or 2 to 4 times greater than the control, as compared to the control. [Claim 61] 19. The probiotic vaccine according to any one of claims 11 to 18, wherein the vaccine continuously produces lysogenic phages. [Claim 62] 19. The probiotic phage according to any one of claims 11 to 18, or the recombinant phage according to any one of claims 12 to 18, wherein -TSGSGSGSGSGSGSG- is used as a linker between the coat protein and the exogenous peptide epitope.

[0025] Other features and advantages of the present invention will become more readily apparent to those of ordinary skill in the art upon review of the following detailed description. [Brief explanation of the drawings]

[0026] [Figure 1A] FIG. 1A shows the physiological effect of oral administration of the probiotic vaccine of the present invention to produce sIgA.

[0027] [Figure 1B] FIG. 1B shows the physiological effects of injecting epitopes targeted via recombinant bacteriophages of the present invention to generate IgA and IgG immune responses.

[0028] [Figure 2A] FIG. 2A shows the results of serum IgA levels for formulations administered as described in Example 9.

[0029] [Figure 2B] FIG. 2B shows the results of serum IgG levels for formulations administered as described in Example 9.

[0030] [Figure 3] FIG. 3 shows the results of serum secretory IgA (SIgA) levels for formulations administered as described in Example 10.

[0031] [Figure 4] FIG. 4 shows the mean tumor volumes resulting from the formulations administered in the prostate cancer xenograft athymic mouse study described in Example 11.

[0032] [Figure 5] FIG. 5 shows the heterologous peptide epitope insertion site in the M13KE-SALV-0004 recombinant phage of Example 1.

[0033] [Figure 6] FIG. 6 shows the heterologous peptide epitope insertion site in the M13KE-SALV-0013 recombinant phage of Example 3. Detailed Description of the Invention

[0034] Provided herein is a probiotic vaccine comprising a recombinant phage (e.g., a filamentous phage), wherein the phage: LPESFDGDPASNTAPLQPEQLQVF (SEQ ID NO: 1; HER2), PESFDGDPASNTAPLQPEQLQ (SEQ ID NO: 2; HER2), QPEKVQTLEAWVIHGG (SEQ ID NO: 3; CD38), ISKRNIQFSCKNIYR (SEQ ID NO: 4; CD38), TFGSVEVHNL (SEQ ID NO: 5; CD38), QTLEA (SEQ ID NO: 6; CD38), IQFSC (SEQ ID NO: 7; CD38), AFPEDRSQPG (SEQ ID NO: 8; PD-1), ALLEIASCL (SEQ ID NO: 9)-IDO1, MYPPPY (SEQ ID NO: 10)-CTLA-4, NPLVA (SEQ ID NO: 11) - claudin, ESKVDPSK (SEQ ID NO: 12) - prostate-specific membrane antigen (PSMA) mimitope, RSSSRTPS (SEQ ID NO: 13) - TNF epitope, AGICAS (SEQ ID NO: 14) - COVID-19 epitope, AG-I-CAS (SEQ ID NO: 15; Ile = "AUA" codon) AKPTAKREAIVKAYG (SEQ ID NO: 16) - monkeypoxvirus, ISPDGCYSL (SEQ ID NO: 17) - monkeypoxvirus, LTFDYVVTF (SEQ ID NO: 18) - monkeypoxvirus, VLYQDVNCT (SEQ ID NO: 19) - Covid Wuhan Spike, QQQQGQTITKKSAAEASKKPRQKRTAVLVQDVGCTFSTVFPVTSFGPLVR (SEQ ID NO: 20) - CovidItaly org9, VLYQGVNCT (SEQ ID NO: 21) - Covid Italy spike, DAPF (SEQ ID NO: 22) - avian influenza, KQIVERILEEE (SEQ ID NO: 23) - avian influenza, RYLTDMTLEE (SEQ ID NO: 24) - avian influenza, DDVKRENPEHEEYNN (SEQ ID NO: 25) - avian influenza, RESGSS (SEQ ID NO: 26) - avian influenza, FRHDSGY (SEQ ID NO: 27) - amyloid-beta AGICASAGICAS (SEQ ID NO: 28) - COVID-19 vaccine ESKVDPSKAW (SEQ ID NO: 29) - prostate-specific membrane antigen (PSMA), DAPFKQIVERILEEE (SEQ ID NO: 30) - avian influenza, RESGSSRYLTDMTLEE (SEQ ID NO: 31) - avian influenza, DDVKRENPEHEEYNN (SEQ ID NO: 32) - avian influenza, CVADYSVLY (SEQ ID NO: 33) - SARS-CoV2, HVGEIPVAY (SEQ ID NO: 34) - SARS-CoV2, NQKLIANQF(NQK-Q8) (SEQ ID NO: 35)-NQK-Q8, SARS-CoV2; NQKLIANAF(NQK-A8) (SEQ ID NO: 36)-NQK-A8, SARS-CoV2, RVAGDSGFAAY (SEQ ID NO: 37) - SARS-CoV2, ESFDGDPASNTAPLQPEQL (SEQ ID NO: 38)-PD-1, EFINVPEGGS (SEQ ID NO: 39) - avian influenza, EFINVPEEFINVPE (SEQ ID NO: 40) - avian influenza, VRSSSRTPSD (SEQ ID NO: 41) - TNF-α inhibitor, RSSSRTPSDK (SEQ ID NO: 42) - TNF-α inhibitor, MYPPPYMYPPPY (SEQ ID NO: 43)-CTLA-4, VELMYPPPYYLGIG (SEQ ID NO: 44)-CTLA-4, LMYPPPYYLGIGN (SEQ ID NO: 45)-CTLA-4, NPLVANPLVA (SEQ ID NO: 46) - Claudin (epithelial ovarian cancer), LPNAIGRGKWWRP (SEQ ID NO: 47) - sclerostin epitope, ETPEGAEA (SEQ ID NO: 48)-TNF-α, KRNIQFSCKNI (SEQ ID NO: 49)-CD38, LTFDY (SEQ ID NO: 50) - monkeypoxvirus, LTFDYLTFDY (SEQ ID NO: 51) - monkeypoxvirus, NTVSCNRPHCITE (SEQ ID NO: 52) - asthma, EERRR (SEQ ID NO: 53) - asthma, EERRREERRR (SEQ ID NO: 54) - asthma, LTHVKIND (SEQ ID NO: 55) - yellow fever, NANPNANPNANPNAN (SEQ ID NO: 56) - malaria, EIRQAGRPNKPDSI (SEQ ID NO: 57)-TL1A, QTLEAQTLEA (SEQ ID NO: 58) - CD38, IQFSCIQFSC (SEQ ID NO: 59) - CD38, DAPFDAPF (SEQ ID NO: 60) - avian influenza, RESGSSRESGSS (SEQ ID NO: 61) - avian influenza, VVVGADVGKS (SEQ ID NO: 62) - KRAS, PTPEEDQILKK (SEQ ID NO: 63)-BTKi, APKAQIKESLR (SEQ ID NO: 64)-PD1, ASPLLGSQHFDVPPRQL (SEQ ID NO: 65)-XPO1, QYDSWPHYPE (SEQ ID NO: 66)-LOXL2, KVIQAQTAFSANPANPAILS (SEQ ID NO: 67) - Syntenin MDA9, PIPHDGNLYPRLYPE (SEQ ID NO: 68) - Syntenin MDA9, SKCRRCRLLD (SEQ ID NO: 69)-CD95, QNTKCRCKPNFFC (SEQ ID NO: 70)-CD95, LCTLAWAHPLVGHM (SEQ ID NO: 71)-IL-23, ESVDPKNYPKK (SEQ ID NO: 72)-IL-1b, YLSETVSNALGPQGRRLEE (SEQ ID NO: 73)-LOXL4, INDPRLRNPSDKFIY (SEQ ID NO: 74)-NMDA1, SPRMSGLLSQTGYLSDNHILI (SEQ ID NO: 75)-DLL3, GLPSLGLTL (SEQ ID NO: 76) - nerve growth receptor, HRGEFS (SEQ ID NO: 77) - nerve growth factor, HRGEFSHRGEFS (SEQ ID NO: 78) - nerve growth factor, THLHWHTV (SEQ ID NO: 79)-β-amyloid, VQDYVQMKASELEQ (SEQ ID NO: 80) - calcitonin, REIEGRVMVTDPENVPEEDG (SEQ ID NO: 81)-PCSK9 (LDL cholesterol), AGAFAPSAAVA (SEQ ID NO: 82)-IL-4, GYYYRDDSLPVLLE (SEQ ID NO: 83) - retinoic acid (acne), EDPAFYMLKGKLQYELQ (SEQ ID NO: 84)-IL-21, ALFDAQAQV (SEQ ID NO: 85)-TYK2, LLYDLTEKQVY (SEQ ID NO: 86)-TYK2, ALDENTKLWYA (SEQ ID NO: 87)-JAK1, VLVESEHQV (SEQ ID NO: 88)-JAK1, APNRTITVDDKMSLRLH (SEQ ID NO: 89)-JAK1, SSDWVTLGTSASLLR (SEQ ID NO: 90) - Ceacam 1, STPYVGLSLSCAANQ (SEQ ID NO: 91) - Ceacam 1, PSPQYSWRINGIPQQHTQVLF (SEQ ID NO: 92) - Ceacam 5, STLYREELMTTLLYYEIFL (SEQ ID NO: 93)-TRPV1, CEKNSVLEVIAYSSSE (SEQ ID NO: 94) - TRPV1 Sub V, NPTTYQMDVNPEGKYS (SEQ ID NO: 95)-EGFR, HTPPLDPQELDILKT (SEQ ID NO: 96) - EGFR, RPKSNIVLLS (SEQ ID NO: 97)-CD-20, SGPKPLFRRMSSLVGPTQ (SEQ ID NO: 98)-CD-20, RPKSNIVLLSAEEKKEQT (SEQ ID NO: 99)-CD-20, DYEDRYYRE (SEQ ID NO: 100) - prion, RPLKGAPLPAL (SEQ ID NO: 101)-ALK, MTEQQWNFAGIEAAASAIQG (SEQ ID NO: 102) - tuberculosis, KTIAYDEEARR (SEQ ID NO: 103) - leprosy, EEFVVEFDLPGIKA (SEQ ID NO: 104) - leprosy, CVTIMTVDNPASTTNKDKLFAVWKITYKDT (SEQ ID NO: 105) - poliovirus 1, ILKEPVHGV (SEQ ID NO: 106)-HIV1, SLYNTVATL (SEQ ID NO: 107) - HIV1, LSEIKGVIVHRLEGV (SEQ ID NO: 108) - measles, MLMAASRAL (SEQ ID NO: 109) - Yersinia pestis, RAMRMVYYL (SEQ ID NO: 110) - Yersinia pestis, FFVRSIGSKGGKLAAGKYTDAVTV (SEQ ID NO: 111) - Yersinia pestis, EEPEESDEDDFGMGGLF (SEQ ID NO: 112) - Leishmania chagasi, CINGVCWTV (SEQ ID NO: 113) - Hepatitis C, KLVALGINAV (SEQ ID NO: 114) - Hepatitis C, NLVPMVATV (SEQ ID NO: 115) - shingles, TPRVTGGGAM (SEQ ID NO: 116) - shingles, TETEPPVAL (SEQ ID NO: 117)-ESRRA, REALHEAL (SEQ ID NO: 118)-ESRRA, DVIGRIENGERL (SEQ ID NO: 119)-TKI, leukemia epitope 2, RLIKEDVYL (SEQ ID NO: 120)-TKI, leukemia epitope 6, KTLETNPYVK (SEQ ID NO: 121)-Ku70, GRADALRVLSK (SEQ ID NO: 122)-IGF-1R, LSRELSSLASQPLSR (SEQ ID NO: 123)-IGF-1R, SLLPDSPVGQ (SEQ ID NO: 124)-IL-23, SPSQPWQRL (SEQ ID NO: 125)-IL-23, FLAEDALNTV (SEQ ID NO: 126) - breast cancer, CTHYMQLAC (SEQ ID NO: 127) - osteoporosis, QEANASAQTKCDIE (SEQ ID NO: 128) - Alaskapoxvirus, VVVGACVGKS (SEQ ID NO: 129) - KRAS, LPAIVREAI (SEQ ID NO: 130) - Dengue virus, SPRRKLISVDSRSVSL (SEQ ID NO: 131)-IL-21, DKPVYTPDQSVKVRV (SEQ ID NO: 132)-C5, and / or DNVLDHLTGRSC (SEQ ID NO: 133) - pertussis; and Bacteria infected with the recombinant phage Also provided is said probiotic vaccine comprising a nucleic acid (eg, a genome) encoding a recombinant polypeptide comprising an exogenous peptide epitope selected from the group consisting of:

[0035] As described herein, the probiotic vaccines of the present invention are particularly useful in methods for preventing, treating, and / or reducing the risk of cancer, autoimmune diseases, respiratory diseases (e.g., asthma), neurological diseases, infectious diseases, and other diseases (e.g., osteoporosis, hypercholesterolemia, etc.). Unlike conventional vaccines, probiotic-based vaccines do not require strict storage conditions and transportation. Conventional live vaccines typically require cold storage before administration. Furthermore, conventional vaccines are developed using attenuated strains of pathogenic bacteria, such as Salmonella, Mycobacterium, and Bacillus. However, these attenuated strains have several drawbacks, including the possibility that they may revert to a virulent form in the body. Furthermore, attenuated vaccines may elicit an additional immune response in individuals, weakening the vaccine's effectiveness. The survival rate of conventional vaccines in the harsh acidic environment of the human intestine is also one factor contributing to the lack of success of conventional vaccines compared to probiotic oral vaccines. Conventional vaccine forms generally cannot withstand the harsh acidic environment and, as a result, cannot reach the mucosal lining. On the other hand, probiotics have excellent adaptability to acidic environments, and their resistance to acid and bile makes them an excellent choice for vaccine development.

[0036] Because SIgA antibodies are present in mucosal areas, while IgA and IgG antibodies are present in the circulation, immune responses within the circulatory system and mucosa differ. Monomeric IgA is present in serum, whereas secretory IgA (SIgA) is present in mucosal secretions. SIgA generally exists in a polymeric form stabilized by linking chains (J chains), particularly as a dimer or tetramer setup, and therefore differs in structure from IgA present in serum. Furthermore, SIgA contains a secretory component (SC) derived from the polymeric Ig receptor (pIgR), which is utilized for crossing epithelial cells during secretion. In humans, there are two subclasses of IgA: IgA1 and IgA2. While the IgA1 subclass predominates in serum, the ratio of IgA1 to IgA2 in mucosal secretions varies depending on the site of production: for example, up to 60% of IgA1 is in saliva, up to 90% in nasal secretions, and up to 60% in intestinal secretions. In human colostrum, approximately 48% of immunoglobulins correspond to the IgA2 subclass and 40% to the IgA1 subclass, conferring adaptations to protect against potentially harmful pathogens as well as a means of controlling the colonization of the neonatal gut microbiota.

[0037] The mucous membranes lining the gastrointestinal, respiratory and urogenital tracts are in constant contact with a wide variety of microorganisms. The gastrointestinal tract (GIT) is home to a large and diverse microbial community, with up to approximately 10 14 The intestinal lining contains microbial cells, composed of approximately 500-1000 species of bacteria, archaea, and fungi. To prevent the invasion of pathogenic microorganisms and control host-bacterial interactions, the mucosal immune system is stimulated to produce SIgA. Mass production of SIgA in the intestine is one of the mechanisms by which the gut-associated lymphoid tissue (GALT) responds to the constant contact of the intestinal mucosa with a large number of diverse microorganisms (mainly bacteria and food-derived antigens).

[0038] Intravenous administration of recombinant phages of the present invention (e.g., MK13E-SALV-0013, etc.) induces IgA and IgG responses, whereas oral administration of the N13 probiotic bacterial vaccine therapy of the present invention (e.g., EcN infected with MK13E-SALV-0013) induces SIgA responses (see Example 10 herein). Current therapeutic monoclonal antibodies lack the ability to act as SIgA and have limited effect on mucosal membranes. Mucosal membranes play an important role in many diseases and serve as a pathway for metastasis.

[0039] As used herein, the term "filamentous" in the context of phage refers to a nonlytic, male-specific bacteriophage that infects Escherichia coli cells carrying the F episome (see 19 for a description). Filamentous phage particles contain a circular, single-stranded DNA genome (positive strand). The phage binds to the bacterial F pilus, and the single-stranded DNA genome then invades the host cell. The dsDNA replicative form is generated by initiating DNA synthesis from the phage's ori (-) structure. Additional cycles of synthesis initiated from the ori (+) structure form single-stranded (+) DNA, which is then packaged into the phage particle without lysing or overtly damaging the host cell.

[0040] As used herein, the term "vaccine" refers to a composition capable of stimulating the immune system of a living organism to provide protection against harmful antigens through prophylaxis or therapy. Preferably, a vaccine or vaccine composition further comprises one or more immunoadjuvant substances.

[0041] As used herein, the terms "preventing," "prevention," "prevention method," or "prevent" generally mean avoiding or minimizing the onset or progression of a disease or condition before it occurs. The term "preventing" encompasses "reducing the likelihood of onset" or "reducing the likelihood of recurrence."

[0042] As used herein, the terms "treat" or "treating," or grammatical variations thereof, refer to achieving one or more of the following: (a) reducing the severity of the disease, (b) inhibiting or preventing the onset of symptoms characteristic of the disease being treated, (c) inhibiting the worsening of symptoms characteristic of the disease being treated, (d) inhibiting or preventing the recurrence of the disease in a subject previously affected by the disease, and (e) inhibiting or preventing the recurrence of symptoms in a subject previously exhibiting symptoms of the disease. Thus, the terms "treating," "treatment," or "treat," or grammatical variations thereof, encompass the reduction, amelioration, or cure of a disease (e.g., cancer, etc.) or condition (or symptoms of the disease or condition) after its onset.

[0043] As used herein, the phrase "reducing risk" refers to prophylactic use to reduce the likelihood of contracting or acquiring a disorder (e.g., cancer or disease) or to limit the onset of a disorder, such as prophylactic administration using the probiotic vaccines and / or recombinant phages of the present invention to avoid contracting a virus or developing cancer, as described herein.

[0044] As used herein, an "effective amount" or "effective dose" refers to an amount that produces a desired effect. For therapeutic purposes, an effective amount is an amount sufficient to produce beneficial or desired clinical results. A preferred effective amount for a particular application can be easily determined by one skilled in the art, taking into account, for example, the subject's physique, age, and weight, the type of disease / disorder to be prevented, treated, and / or reduced, and the time elapsed since the onset of the disease / disorder. In the present invention, from the perspective of prevention, treatment, and / or risk reduction, an effective amount of a composition is an amount sufficient to induce a humoral immune response and / or a cellular immune response against the disease / disorder.

[0045] According to various aspects and embodiments of the invention described herein, a "subject" or "host" preferably refers to a mammal, most preferably a human, who may be suffering from, suspected of suffering from, or at risk of suffering from cancer and / or an infectious disease (such as, for example, Korean fever virus infection).

[0046] Exogenous peptide epitopes As used herein, the phrase "exogenous peptide epitope" refers to a peptide or sequence that is not native or naturally occurring in the host phage strain used to produce the recombinant phage of the present invention. In other words, the exogenous peptide epitope and the nucleic acid encoding it are heterologous (i.e., foreign) to the peptide sequence of the particular phage strain being utilized. In certain embodiments, the length of the exogenous peptide epitope can be in a range selected from the group consisting of 4 to 20 amino acids, 4 to 25 amino acids, 5 to 20 amino acids, 5 to 25 amino acids, 5 to 30 amino acids, 5 to 35 amino acids, 5 to 40 amino acids, 5 to 45 amino acids, and 5 to 50 amino acids. In certain embodiments, the length of the exogenous peptide epitope is in the range of 5 to 20 amino acids.

[0047] According to the present invention, an exogenous peptide epitope (e.g., an antigenic peptide) for use in the recombinant phage vaccines and / or probiotic vaccines provided herein comprises or consists of an amino acid sequence set forth in any one of SEQ ID NOs: 9 to 133, or a fragment or variant thereof.

[0048] The epitopes used to culture the recombinant phage with the probiotic bacterial cells will vary depending on the type of disease (eg, cancer) to be treated or prevented, or the infectious disease to be prevented.

[0049] For example, in certain embodiments, the following exogenous peptide epitopes can be used to prevent, treat, and / or reduce the risk of cancer, including multiple myeloma, epithelial cancer, epithelial ovarian cancer, mucosal melanoma, non-small cell lung cancer, melanoma, head and neck cancer, renal cell carcinoma, Hodgkin's lymphoma, cutaneous squamous cell carcinoma, glioblastoma, esophageal cancer, gastric cancer, duodenal cancer, small intestine cancer, appendix cancer, colon cancer, rectal cancer, colorectal cancer, anal cancer, pancreatic cancer, liver cancer, gallbladder cancer, spleen cancer, kidney cancer, bladder cancer, prostate cancer, testicular cancer, uterine cancer, endometrial cancer, ovarian cancer, vaginal cancer, vulvar cancer, breast cancer, lung cancer, thyroid cancer, thymus cancer, brain cancer, nervous system cancer, glioma, oral cancer, skin cancer, blood cancer, and lymphoma. , eye cancer, bone cancer, bone marrow cancer, muscle cancer, non-small cell lung cancer (NSCLC), head and neck squamous cell carcinoma (HNSCC), urothelial cancer, non-muscle-invasive bladder cancer (NMIBC), colon or rectal cancer, esophageal cancer or certain gastroesophageal junction (GEJ) cancer, cervical cancer, renal cell carcinoma (RCC), advanced endometrial cancer, cutaneous squamous cell carcinoma (cSCC) and / or triple-negative breast cancer (TNBC); and the exogenous peptide epitope is selected from SEQ ID NOs: 9-12, 29, 38, 43-46, 49, 58, 59, 62-70, 73, 75, 84, 90-92, 95, 96, 101, 117-123, 126, 129 and / or 131.

[0050] In another embodiment, the exogenous peptide epitope comprises no more than 5, no more than 4, no more than 3, no more than 2, or 1 amino acid substitution within SEQ ID NOs: 9-12, 29, 38, 43-46, 49, 58, 59, 62-70, 73, 75, 84, 90-92, 95, 96, 101, 117-123, 126, 129, and / or 131.

[0051] As used herein, the term "fragment," or grammatical variations thereof, refers to a smaller peptide (e.g., a subset of amino acids) compared to the complete reference peptide sequence from which it is derived. For example, a fragment may have 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more fewer amino acids than the complete reference peptide.

[0052] As used herein, the terms "variant thereof," "variant," or grammatical variations thereof refer to a biologically active polypeptide having at least about 80% amino acid sequence identity with a reference sequence polypeptide, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity, without considering any conservative substitutions as part of the sequence identity. Such variants include, for example, polypeptides in which one or more amino acid residues are added or deleted at the N- or C-terminus of the polypeptide. In some embodiments, variants have at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, and 99% amino acid sequence identity. In some embodiments, variants have at least about 85% amino acid sequence identity. In some embodiments, variants have at least about 90% amino acid sequence identity. In some embodiments, variants have at least about 95% amino acid sequence identity with a native sequence polypeptide.

[0053] As used herein, "percent (%) amino acid sequence identity" and "homology" with respect to peptide or polypeptide sequences are defined as the percentage of amino acid residues in a candidate sequence that are identical to amino acid residues in a particular peptide or polypeptide sequence after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity, without considering any conservative substitutions as part of the sequence identity. Alignment to determine percent amino acid sequence identity can be performed using a variety of methods known to those of skill in the art, for example, publicly available computer software such as BLAST, BLAST-2, ALIGN, or MEGALIGN™ (DNASTAR) software. Those skilled in the art can determine appropriate parameters for measuring alignment, including the algorithms necessary to achieve maximal alignment over the entire length of the sequences being compared.

[0054] In certain embodiments, amino acid substitutions include, but are not limited to, substituting one amino acid for another in a polypeptide. Examples of conservative substitutions are shown in Table 1. In another embodiment, conservative amino acid substitutions can be introduced into a peptide of interest and the resulting product screened for a desired activity, such as improved immunogenicity.

[0055] In some embodiments, the modification may involve introducing at least one conservative substitution into a peptide or protein, in which at least one characteristic of the amino acid, such as size, shape, charge, etc., is preserved. A "conservative amino acid substitution" refers to a substitution with a structurally and / or functionally similar amino acid that does not substantially change the function of the protein. Examples of conservative substitutions include replacing an amino acid from any of the following groups with another amino acid from the same group (U.S. Pat. No. 5,767,063; Kyte and Doolittle, J. Mol. Biol. 157: 105-132 (1982)): (1) Hydrophobic: Norleucine, Met, Ala, Val, Leu, Ile; (2) Neutral hydrophilic: Cys, Ser, Thr, Asn, Gln; (3) Acidic: Asp, Glu; (4) basic: His, Lys, Arg; (5) residues that influence chain orientation: Gly, Pro; (6) Aromatic: Trp, Tyr, Phe [Table 1]

[0056] Substitutions can be made, for example, by changing Val to Leu, Ser to Thr, or Asp to Glu. Other substitutions may also be considered conservative, depending on the specific amino acid's environment and role in the protein's three-dimensional structure. For example, Glu can be substituted with Gln or Asp with Asn to change the pKa while maintaining the size and structure of the amino acid side chain.

[0057] In some embodiments, the amino acid residue selected for modification (e.g., substitution with another amino acid residue) is selected from the group consisting of His, Glu, Asp, Cys, Lys, and Tyr. In some embodiments, the amino acid residues selected for modification include His and Glu residues. These amino acid residues typically have pKa values ​​(even in whole proteins) between about pH 6 and about pH 8 and therefore have high buffering capacity. In some embodiments, at least one conservative amino acid substitution is selected from the group consisting of His to Arg, Glu to Gln, Asp to Asn, Lys to Arg, and Tyr to Phe.

[0058] In some embodiments, at least one of the one or more amino acid modifications comprises an amino acid substitution with an alanine residue. Substituting or changing an amino acid residue with a high buffer capacity to an alanine residue can be advantageous in applications where it is desirable to reduce the buffer capacity of a peptide or protein. Note that norleucine is not encoded and is modified after substitution.

[0059] bacteria As mentioned above, the present invention provides a probiotic vaccine comprising a recombinant phage and a bacterium infected with the recombinant phage. In a specific embodiment, the recombinant phage is capable of producing F factor (F), which produces pili for conjugation with other bacteria and sex pili. + Any bacteria having a sex factor (e.g., a bacterial cell) is suitable for use in the present invention. + The cells are F - When mated with a cell, two F + The cells produce and transfer the plasmid to other F - It can be delivered to cells. +The pili on the cell interact with the recipient cell, allowing the formation of a mating junction, which causes a break in one strand of DNA that is then unwound and transferred to the recipient cell. Examples of F factor-containing bacteria include E. coli Nissle 1917 and E. coli ER2738.

[0060] Thus, in certain embodiments, examples of probiotic bacteria for use in the present invention include, but are not limited to, Escherichia coli (E. coli) Nissle 1917 strain, Escherichia coli (E. coli) ER2738 strain, Bacillus coagulans (Bacillus coagulans) GBI-30,6086 strain, Bifidobacterium animalis subsp. lactis (Bifidobacterium animalis) BB-12 strain, Bifidobacterium longum subsp. infantis (Bifidobacterium longum) infantis (Bifidobacterium longum) Lactobacillus acidophilus (Lactobacillus acidophilus) NCFM strain, Lactobacillus paracasei (Lactobacillus paracasei) Stl 1 strain (or NCC2461 strain), Lactobacillus johnsonii (Lactobacillus Lactobacillus johnsonii Lai strain (also called Lactobacillus LCI strain, Lactobacillus johnsonii NCC533 strain), Lactobacillus plantarum 299v strain, Lactobacillus reuteri ATCC 55730 strain (Lactobacillus reuteri SD2112 strain), Lactobacillus reuteri Protectis strain (DSM 17938 strain, daughter strain of ATCC 55730 strain), Lactobacillus reuteri Prodentis strain (DSM 17938 / ATCC 55730 and ATCC PTA5289 strain), Lactobacillus rhamnosus GG strain, Saccharomyces boulardii, Lactobacillus rhamnosus GR-1 strain and Lactobacillus reuteria mixture of Lactobacillus acidophilus NCFM and Bifidobacterium bifidum BB-12; a mixture of Lactobacillus acidophilus CL1285 and Lactobacillus casei LBC80R; a mixture of Lactobacillus plantarum HEAL 9 and Lactobacillus paracasei 8700:2; a mixture of Lactobacillus bulgaricus, Lactococcus thermophiles, and / or Lactobacillus bifidus In some embodiments, the probiotic bacteria used in the present disclosure may be a mixture of any two or more of the strains described herein above. In certain embodiments, the probiotic bacteria are Escherichia coli Nissle 1917 or Escherichia coli ER2738. In another embodiment, the probiotic bacteria are Escherichia coli Nissle 1917. In yet another embodiment, the probiotic bacteria are Escherichia coli ER2738.

[0061] In another embodiment, when the peptide is displayed as part of pVIII, suitable phage systems include type 8, type 88 (also referred to herein as f88), and type 8+8, as described herein (e.g., Zhong et al., J. Biol. Chem. 269:24183-24188, 1994; Smith and Scott, Methods in Enzymology, 217:228-257, 1993; Smith, Gene, 128:1-2, 1993, each of which is incorporated herein by reference in its entirety). When pIII is utilized, suitable phage systems include type 3, type 33, and type 3+3. When the peptide is inserted into pVI, suitable phage systems include type 6, type 66, and type 6+6. Additionally, phage T7 and phage 8 vector systems can be used. In one particular embodiment, the peptides of the library are expressed fused to coat proteins of a filamentous bacteriophage, such that the peptides are expressed on the surface of the virion and available to interact with target molecules or cell surface receptors.

[0062] In a specific embodiment, the f88-4 library is constructed using the filamentous phage display vector f88-4 (GenBank accession # AF218363), in which selected peptides are fused to the pVIII coat protein. The PVIII protein is the major coat protein of phages and is present at approximately 2700 copies per phage. PVIII is synthesized as a precoat protein containing a 23-amino acid leader peptide, which is cleaved to yield a mature 50-residue transmembrane protein. Two systems that allow for the generation of mosaic phages have been used: the "Type 8+8" system and the "Type 88" system (Smith GP (1993) Surface display and peptide libraries. Gene, 128, 1-2, incorporated herein by reference in their entirety). The "Type 88" (e.g., f88-4) system has the advantage of containing two pVIII genes in the same infectious phage genome. This eliminates the need for multiple helper phages or superinfections, which would otherwise produce only one type of phage.

[0063] In another embodiment, bacteria suitable for use in the present invention are small (typical linear dimensions of about 1 μm), non-compartmentalized organisms with at least one circular DNA chromosome and 70S ribosomes. As used herein, the term "bacteria" encompasses all variants of bacteria (e.g., endogenous bacteria naturally occurring in closed systems, environmental bacteria, or bacteria released for bioremediation or other efforts).

[0064] In the probiotic vaccines provided herein, the recombinant phage of the present invention is introduced into (e.g., infects) donor bacteria (e.g., probiotic bacteria and / or resident bacteria) and delivered to a subject (e.g., a human subject) in need thereof. For example, probiotic bacteria are live bacteria that provide a health benefit to the host and / or are at least harmless (e.g., non-pathogenic) to the host, such as a human patient. Thus, embodiments of the present invention contemplate using donor bacteria (e.g., probiotic bacteria and / or resident bacteria) that have been introduced or infected with a non-lytic or inducible bacteriophage that can express and produce bacteriophage-based delivery particles in situ. Probiotics have been found to penetrate the inner mucosal layer of the intestine, not only aiding in the invasion of harmful bacteria but also preventing many enteric infections.

[0065] In certain embodiments, examples of probiotic bacteria for use in the present invention include those listed in the "PROBIO" database (" / / bidd.group / probio / homepage.htm"; Shamekhi et al., Clin Transl Oncol (2020) 22(8):1227-39. doi:10.1007 / s12094-019-02270-0), which is incorporated by reference in its entirety for all purposes. This database includes 329 probiotics currently on the market and 115 probiotic bacteria in clinical trials. In a specific embodiment, the recombinant phage of the present invention is selected from the group consisting of Bacillus amyloliquefaciens, Bacillus polyfermenticus strain Bispan, Bifidobacterium animalis subsp. lactis strain BB-12, Bifidobacterium animalis subsp. lactis strain GPS1209, Bifidobacterium animalis subsp.Lactobacillus acidophilus strain HN019 (DR1064), Bifidobacterium bifidum strain BB-12, Bifidobacterium bifidum strain Rosell-71, Bifidobacterium breve strain M-16V, Bifidobacterium longum, Bifidobacterium thermophilum, Lactobacillus acidophilus strain La-1, Lactobacillus brevis strain HA-112, Lactobacillus fermentum The subject is infected with or introduced into a specific probiotic bacterium having immunomodulatory activity, selected from the group consisting of Lactobacillus helveticus Lafti L10 strain, Lactobacillus helveticus Rosell-52 strain, Lactobacillus paracasei Lafti L26 strain, Lactobacillus paracasei subsp. paracasei strain 431, Lactobacillus rhamnosus HN001(DR20) strain, Streptococcus salivarius DSM 13084 strain, and Streptococcus thermophilus. See, e.g., Table 1 in Singh et al., Front Immunol. 2022; 13: 1002674; Published online 2022 Oct 3. doi: 10.3389 / fimmu.2022.1002674, which is incorporated by reference in its entirety for all purposes.

[0066] In another embodiment, the recombinant bacteriophage of the present disclosure is capable of inhibiting Bacteroides thetaiotamicron (e.g., strain Bl), Bacteroides fragilis (e.g., strain ATCC 51477-B1, strain B40-8, strain Bf-1), Bacteroides cassae (e.g., strain phiHSCl), Bacteroides ovatus (e.g., strain phiHSC02), Clostridium difficile (e.g., strains phiC2, phiC5, phiC6, phiC8, phiCD119, phiCD27), Klebsiella pneumoniae (e.g., strain KP01K2, Kl), or any of the following: Bacteroides thetaiotamicron (e.g., strain Bl), Bacteroides fragilis (e.g., strain ATCC 51477-B1, strain B40-8, strain Bf-1), Bacteroides caccae (e.g., strain phiHSCl), Bacteroides ovatus (e.g., strain phiHSC02), Clostridium difficile (e.g., strains phiC2, phiC5, phiC6, phiC8, phiCD119, phiCD27), Klebsiella pneumoniae (e.g., strain KP01K2, Kl), or any of the following: Bacteroides thetaiotamicron (e.g., strain Bl), Bacteroides fragilis (e.g., strain ATCC 51477-B1, strain B40-8, strain Bf-1), Bacteroides thetaiotamicron (e.g., strain Bl), Bacteroides thetaiotamicron (e.g., strain Bl), Bacteroides thetaiotamicron (e.g., strain Bl), Bactero It is intended to target bacteria other than E. coli, including Staphylococcus aureus (e.g., phiNMl, Kpn5, KP34, JDOOl), Staphylococcus aureus (e.g., phiNMl, 80α), Enterococcus faecalis (e.g., IME-EF1), Enterococcus faecium (e.g., ENB6, C33), and Pseudomonas aeruginosa (e.g., phiKMV, PAK-P1, LKD16, LKA1, δ-1, σ-1, Jl).

[0067] Thus, the bacteriophages of the present disclosure can target (e.g., specifically target) bacterial cells belonging to any one or more of the bacterial genera and / or species listed above, as well as other bacterial cells and microorganisms.

[0068] As used herein, "endogenous" bacterial cells may refer to non-pathogenic bacteria that are part of the normal internal ecosystem, such as the bacterial flora.

[0069] In yet another embodiment, the bacterial cell of the present disclosure is an anaerobic bacterial cell (e.g., a cell that does not require oxygen for growth). Anaerobic bacterial cells include, for example, facultative anaerobic cells such as Escherichia coli, Shewanella oneidensis, and Listeria monocytogenes. Anaerobic bacterial cells also include obligate anaerobic cells such as Bacteroides species and Clostridium species. For example, in humans, anaerobic bacterial cells are most abundant in the gastrointestinal tract. Thus, the bacteriophage of the present disclosure can target (e.g., specifically target) anaerobic bacterial cells.

[0070] Recombinant bacteriophage Provided herein is a recombinant bacteriophage (e.g., filamentous) comprising a recombinant phage genome, or a fragment or variant thereof, comprising a nucleic acid encoding a polypeptide comprising an exogenous peptide epitope, LPESFDGDPASNTAPLQPEQLQVF (SEQ ID NO: 1; HER2), PESFDGDPASNTAPLQPEQLQ (SEQ ID NO: 2; HER2), QPEKVQTLEAWVIHGG (SEQ ID NO: 3; CD38), ISKRNIQFSCKNIYR (SEQ ID NO: 4; CD38), TFGSVEVHNL (SEQ ID NO: 5; CD38), QTLEA (SEQ ID NO: 6; CD38), IQFSC (SEQ ID NO: 7; CD38), AFPEDRSQPG (SEQ ID NO: 8; PD-1), ALLEIASCL (SEQ ID NO: 9)-IDO1, MYPPPY (SEQ ID NO: 10)-CTLA-4, NPLVA (SEQ ID NO: 11) - claudin, ESKVDPSK (SEQ ID NO: 12) - prostate-specific membrane antigen (PSMA) mimitope, RSSSRTPS (SEQ ID NO: 13) - TNF epitope, AGICAS (SEQ ID NO: 14) - COVID-19 epitope, AG-I-CAS (SEQ ID NO: 15; Ile = "AUA" codon) AKPTAKREAIVKAYG (SEQ ID NO: 16) - monkeypoxvirus, ISPDGCYSL (SEQ ID NO: 17) - monkeypoxvirus, LTFDYVVTF (SEQ ID NO: 18) - monkeypoxvirus, VLYQDVNCT (SEQ ID NO: 19) - Covid Wuhan Spike, QQQQGQTITKKSAAEASKKPRQKRTAVLVQDVGCTFSTVFPVTSFGPLVR (SEQ ID NO: 20) - CovidItaly org9, VLYQGVNCT (SEQ ID NO: 21) - Covid Italy spike, DAPF (SEQ ID NO: 22) - avian influenza, KQIVERILEEE (SEQ ID NO: 23) - avian influenza, RYLTDMTLEE (SEQ ID NO: 24) - avian influenza, DDVKRENPEHEEYNN (SEQ ID NO: 25) - avian influenza, RESGSS (SEQ ID NO: 26) - avian influenza, FRHDSGY (SEQ ID NO: 27) - amyloid-beta AGICASAGICAS (SEQ ID NO: 28) - COVID-19 vaccine ESKVDPSKAW (SEQ ID NO: 29) - prostate-specific membrane antigen (PSMA), DAPFKQIVERILEEE (SEQ ID NO: 30) - avian influenza, RESGSSRYLTDMTLEE (SEQ ID NO: 31) - avian influenza, DDVKRENPEHEEYNN (SEQ ID NO: 32) - avian influenza, CVADYSVLY (SEQ ID NO: 33) - SARS-CoV2, HVGEIPVAY (SEQ ID NO: 34) - SARS-CoV2, NQKLIANQF(NQK-Q8) (SEQ ID NO: 35)-NQK-Q8, SARS-CoV2; NQKLIANAF(NQK-A8) (SEQ ID NO: 36)-NQK-A8, SARS-CoV2, RVAGDSGFAAY (SEQ ID NO: 37) - SARS-CoV2, ESFDGDPASNTAPLQPEQL (SEQ ID NO: 38)-PD-1, EFINVPEGGS (SEQ ID NO: 39) - avian influenza, EFINVPEEFINVPE (SEQ ID NO: 40) - avian influenza, VRSSSRTPSD (SEQ ID NO: 41) - TNF-α inhibitor, RSSSRTPSDK (SEQ ID NO: 42) - TNF-α inhibitor, MYPPPYMYPPPY (SEQ ID NO: 43)-CTLA-4, VELMYPPPYYLGIG (SEQ ID NO: 44)-CTLA-4, LMYPPPYYLGIGN (SEQ ID NO: 45)-CTLA-4, NPLVANPLVA (SEQ ID NO: 46) - Claudin (epithelial ovarian cancer), LPNAIGRGKWWRP (SEQ ID NO: 47) - sclerostin epitope, ETPEGAEA (SEQ ID NO: 48)-TNF-α, KRNIQFSCKNI (SEQ ID NO: 49)-CD38, LTFDY (SEQ ID NO: 50) - monkeypoxvirus, LTFDYLTFDY (SEQ ID NO: 51) - monkeypoxvirus, NTVSCNRPHCITE (SEQ ID NO: 52) - asthma, EERRR (SEQ ID NO: 53) - asthma, EERRREERRR (SEQ ID NO: 54) - asthma, LTHVKIND (SEQ ID NO: 55) - yellow fever, NANPNANPNANPNAN (SEQ ID NO: 56) - malaria, EIRQAGRPNKPDSI (SEQ ID NO: 57)-TL1A, QTLEAQTLEA (SEQ ID NO: 58) - CD38, IQFSCIQFSC (SEQ ID NO: 59) - CD38, DAPFDAPF (SEQ ID NO: 60) - avian influenza, RESGSSRESGSS (SEQ ID NO: 61) - avian influenza, VVVGADVGKS (SEQ ID NO: 62) - KRAS, PTPEEDQILKK (SEQ ID NO: 63)-BTKi, APKAQIKESLR (SEQ ID NO: 64)-PD1, ASPLLGSQHFDVPPRQL (SEQ ID NO: 65)-XPO1, QYDSWPHYPE (SEQ ID NO: 66)-LOXL2, KVIQAQTAFSANPANPAILS (SEQ ID NO: 67) - Syntenin MDA9, PIPHDGNLYPRLYPE (SEQ ID NO: 68) - Syntenin MDA9, SKCRRCRLLD (SEQ ID NO: 69)-CD95, QNTKCRCKPNFFC (SEQ ID NO: 70)-CD95, LCTLAWAHPLVGHM (SEQ ID NO: 71)-IL-23, ESVDPKNYPKK (SEQ ID NO: 72)-IL-1b, YLSETVSNALGPQGRRLEE (SEQ ID NO: 73)-LOXL4, INDPRLRNPSDKFIY (SEQ ID NO: 74)-NMDA1, SPRMSGLLSQTGYLSDNHILI (SEQ ID NO: 75)-DLL3, GLPSLGLTL (SEQ ID NO: 76) - nerve growth receptor, HRGEFS (SEQ ID NO: 77) - nerve growth factor, HRGEFSHRGEFS (SEQ ID NO: 78) - nerve growth factor, THLHWHTV (SEQ ID NO: 79)-β-amyloid, VQDYVQMKASELEQ (SEQ ID NO: 80) - calcitonin, REIEGRVMVTDPENVPEEDG (SEQ ID NO: 81)-PCSK9 (LDL cholesterol), AGAFAPSAAVA (SEQ ID NO: 82)-IL-4, GYYYRDDSLPVLLE (SEQ ID NO: 83) - retinoic acid (acne), EDPAFYMLKGKLQYELQ (SEQ ID NO: 84)-IL-21, ALFDAQAQV (SEQ ID NO: 85)-TYK2, LLYDLTEKQVY (SEQ ID NO: 86)-TYK2, ALDENTKLWYA (SEQ ID NO: 87)-JAK1, VLVESEHQV (SEQ ID NO: 88)-JAK1, APNRTITVDDKMSLRLH (SEQ ID NO: 89)-JAK1, SSDWVTLGTSASLLR (SEQ ID NO: 90) - Ceacam 1, STPYVGLSLSCAANQ (SEQ ID NO: 91) - Ceacam 1, PSPQYSWRINGIPQQHTQVLF (SEQ ID NO: 92) - Ceacam 5, STLYREELMTTLLYYEIFL (SEQ ID NO: 93)-TRPV1, CEKNSVLEVIAYSSSE (SEQ ID NO: 94) - TRPV1 Sub V, NPTTYQMDVNPEGKYS (SEQ ID NO: 95)-EGFR, HTPPLDPQELDILKT (SEQ ID NO: 96) - EGFR, RPKSNIVLLS (SEQ ID NO: 97)-CD-20, SGPKPLFRRMSSLVGPTQ (SEQ ID NO: 98)-CD-20, RPKSNIVLLSAEEKKEQT (SEQ ID NO: 99)-CD-20, DYEDRYYRE (SEQ ID NO: 100) - prion, RPLKGAPLPAL (SEQ ID NO: 101)-ALK, MTEQQWNFAGIEAAASAIQG (SEQ ID NO: 102) - tuberculosis, KTIAYDEEARR (SEQ ID NO: 103) - leprosy, EEFVVEFDLPGIKA (SEQ ID NO: 104) - leprosy, CVTIMTVDNPASTTNKDKLFAVWKITYKDT (SEQ ID NO: 105) - poliovirus 1, ILKEPVHGV (SEQ ID NO: 106)-HIV1, SLYNTVATL (SEQ ID NO: 107) - HIV1, LSEIKGVIVHRLEGV (SEQ ID NO: 108) - measles, MLMAASRAL (SEQ ID NO: 109) - Yersinia pestis, RAMRMVYYL (SEQ ID NO: 110) - Yersinia pestis, FFVRSIGSKGGKLAAGKYTDAVTV (SEQ ID NO: 111) - Yersinia pestis, EEPEESDEDDFGMGGLF (SEQ ID NO: 112) - Leishmania chagasi, CINGVCWTV (SEQ ID NO: 113) - Hepatitis C, KLVALGINAV (SEQ ID NO: 114) - Hepatitis C, NLVPMVATV (SEQ ID NO: 115) - shingles, TPRVTGGGAM (SEQ ID NO: 116) - shingles, TETEPPVAL (SEQ ID NO: 117)-ESRRA, REALHEAL (SEQ ID NO: 118)-ESRRA, DVIGRIENGERL (SEQ ID NO: 119)-TKI, leukemia epitope 2, RLIKEDVYL (SEQ ID NO: 120)-TKI, leukemia epitope 6, KTLETNPYVK (SEQ ID NO: 121)-Ku70, GRADALRVLSK (SEQ ID NO: 122)-IGF-1R, LSRELSSLASQPLSR (SEQ ID NO: 123)-IGF-1R, SLLPDSPVGQ (SEQ ID NO: 124)-IL-23, SPSQPWQRL (SEQ ID NO: 125)-IL-23, FLAEDALNTV (SEQ ID NO: 126) - breast cancer, CTHYMQLAC (SEQ ID NO: 127) - osteoporosis, QEANASAQTKCDIE (SEQ ID NO: 128) - Alaskapoxvirus, VVVGACVGKS (SEQ ID NO: 129) - KRAS, LPAIVREAI (SEQ ID NO: 130) - Dengue virus, SPRRKLISVDSRSVSL (SEQ ID NO: 131)-IL-21, DKPVYTPDQSVKVRV (SEQ ID NO: 132)-C5, and / or DNVLDHLTGRSC (SEQ ID NO: 133) - pertussis Also provided is a recombinant bacteriophage selected from the group consisting of:

[0071] In certain embodiments of the recombinant phage, the exogenous peptide epitope is functionally expressed on a coat protein of the phage selected from the group consisting of pIII, pVI, pVII, pVIII, and pIX. In certain embodiments of the recombinant phage, the coat protein is pIII. In another embodiment, the coat protein is pVIII. As used herein, "functionally expressed" refers to expression such that the exogenous peptide epitope is displayed on the surface of the phage and is capable of eliciting an immune response.

[0072] In further embodiments, the phage is selected from the group consisting of M13, fd, IKe, CTX-φ, Pf1, Pf2, Pf3, f1, MKE, M13KE, type 8, type f88, f88-4, Myoviridae (P1-like viruses, P2-like viruses, Mu-like viruses, SPO1-like viruses, phiH-like viruses), Siphoviridae (λ-like viruses, γ-like viruses, T1-like viruses, T5-like viruses, c2-like viruses, L5-like viruses, psiM1-like viruses, phiC31-like viruses, N15-like viruses), Podoviridae (phi29-like viruses, P22-like viruses, N4-like viruses), Tectiviridae (tectiviruses), Corticoviridae (corticoviruses), In yet another embodiment, the phage is selected from the group consisting of filamentous phage including M13, fd, IKe, CTX-φ, Pf1, Pf2, Pf3, f1, MKE, M13KE, Type 8, Type f88, and f88-4.

[0073] In a specific embodiment, the phage is M13KE. In yet another embodiment, the phage is M13KE and the insertion site is between the pill signal peptide and the pill protein coding sequence, as shown in Figure 5. In another embodiment, the phage is f88 or f88-4. In yet another embodiment, the phage is f88 or f88-4 and the insertion site is between the signal peptide and the pVIII coding sequence, as shown in Example 12. In yet another embodiment, the recombinant phage produces IgG antibodies that bind to the exogenous peptide epitope.

[0074] In certain embodiments of the present disclosure, nucleic acids encoding exogenous peptide epitopes are recombinantly delivered to a population of microbial cells, e.g., probiotic bacterial cells, by recombinantly incorporating naturally occurring, engineered (e.g., rationally engineered), or adaptively evolved bacteriophages. Bacteriophages, or phages, are viruses that infect and replicate in bacteria. Bacteriophages are composed of proteins encompassing a DNA or RNA genome and can have relatively simple or complex structures. Their genomes can encode as few as four genes or as many as several hundred genes. After injecting their genomes into the cytoplasm, bacteriophages replicate within the bacteria, replicating using either a lytic cycle, which lyses the bacterial cell, or a lysogenic (non-lytic) cycle, which leaves the bacterial cell intact. In some embodiments, the bacteriophages of the present disclosure are non-lytic (also referred to as lysogenic or temperate phages). Non-lytic phages also include phages that are actively secreted from infected cells without lysis, such as, but not limited to, filamentous phages such as f1, M13, M13KE, type 8, type f88, f88-4, fd, IKe, CTX-φ, Pf1, Pf2, and Pf3. Thus, after delivery of an exogenous peptide epitope into a bacterial cell by a phage, the bacterial cell can survive and stably maintain expression of the antigenic epitope. In some embodiments, lytic bacteriophages can be used as delivery vehicles. When used with phagemid systems, lytic phages essentially function as cargo shuttles and do not essentially lyse target cells.

[0075] Examples of non-lytic bacteriophages that may be used in the present invention include, but are not limited to, Myoviridae (P1-like viruses, P2-like viruses, Mu-like viruses, SPO1-like viruses, phiH-like viruses), Siphoviridae (λ-like viruses, γ-like viruses, T1-like viruses, T5-like viruses, c2-like viruses, L5-like viruses, psiM1-like viruses, phiC31-like viruses, N15-like viruses), Podoviridae (phi29-like viruses, P22-like viruses, N4-like viruses), Tectiviridae (tectiviruses), Corticoviridae (corticoviruses), Liposthrixviridae (alf-like viruses), and the like. Examples of suitable phages include those selected from the group consisting of: Aliphothrixvirus, Betaliphothrixvirus, Gammaliphothrixvirus, Deltaliphothrixvirus), Plasmaviridae (plasma viruses), Rudiviridae (rudivruria), Fuselloviridae (fuselloviruses), Inoviridae (inoviruses, plectroviruses, M13-like viruses, fd-like viruses), Microviridae (microviruses, spiromicroviruses, bdelomicroviruses, chlamydiamicroviruses), Leviviridae (leviviruses, alloleviviruses), and Cystoviridae (cystoviruses). Such phages may be naturally occurring or artificially engineered. In some embodiments, the bacteriophage is a coliphage (e.g., capable of infecting Escherichia coli). Those skilled in the art will readily appreciate that other bacteriophages can be used in accordance with the present disclosure.

[0076] In a specific embodiment, the bacteriophage used in the present invention is a filamentous phage. Filamentous phages constitute a large family of bacterial viruses that infect many Gram-negative bacteria. Suitable known filamentous phages include, for example, phages that infect Escherichia coli, such as f1, M13KE, type 8, type f88, f88-4, fd, and Ike. f1, M13, and fd phages have been used for filamentous phage display. Their genomes are over 98% identical, and their gene products are interchangeable.

[0077] Unlike the assembly of many other bacteriophages, the assembly of filamentous phage has the unique feature of being a secretory process. Incorporation of the coat polypeptide into the growing phage occurs within the cytoplasmic membrane, and the nascent phage is exported to the outside of the cell once assembled. Escherichia coli does not lyse during this process. It is well known to those skilled in the art that five viral coat proteins (pIII, pVI, pVII, pVIII, and pIX) are inserted into the cytoplasmic membrane before being incorporated into phage particles. For example, the majority of pIII translocates across the membrane into the periplasm, while the C-terminal hydrophobic tail anchors the protein to the membrane.

[0078] In certain embodiments, the bacteriophage used to prepare the recombinant phage of the present invention is M13 bacteriophage (e.g., M13KE, catalog number 8101S, manufactured by New England Biolabs). M13 is a filamentous bacteriophage belonging to the Inoviridae family and is composed of a circular, single-stranded DNA. M13 phage is approximately 900 nm long, 6-7 nm in diameter, and contains five proteins. The minor coat protein, P3, binds to a receptor at the tip of the F pilus of an Escherichia coli host cell. Therefore, in certain embodiments, the probiotic vaccines of the present invention and their use in the methods of preventing, treating, and / or reducing the risk of diseases of the present invention comprise delivering to bacterial cells a recombinant M13 bacteriophage of the present invention modified to functionally express at least one exogenous peptide epitope selected from SEQ ID NOs: 9-133 in any of the coat proteins selected from the group consisting of pIII, pVI, pVII, pVIII, and pIX. In certain embodiments, the exogenous peptide epitope is functionally expressed in the pIII gene of M13 bacteriophage. In certain embodiments, the bacteriophage is M13KE. In yet another embodiment, the phage is M13KE and the insertion site is between the pIII signal peptide and the pIII protein coding sequence, as shown in Figure 5.

[0079] In another embodiment, the probiotic vaccines of the present invention and their use in the methods of preventing, treating, and / or reducing the risk of diseases of the present invention comprise delivering to a bacterial cell a recombinant f88 bacteriophage of the present invention modified to functionally express at least one exogenous peptide epitope selected from SEQ ID NOs: 9-133 in any of its coat proteins selected from the group consisting of pIII, pVI, pVII, pVIII, and pIX. In a specific embodiment, the exogenous peptide epitope is functionally expressed in the pVIII gene of the f88 bacteriophage. In a specific embodiment, the bacteriophage is f88-4 (GenBank Accession # AF218363). In yet another embodiment, the phage is f88 or f88-4, and the insertion site is between the signal peptide and the pVIII coding sequence, as shown in Example 12.

[0080] Also provided is a method for producing the recombinant phage of the present invention, which comprises expressing in a bacterium a recombinant phage genome comprising a nucleic acid encoding a polypeptide comprising an exogenous peptide epitope selected from the group consisting of SEQ ID NOs: 9 to 133.

[0081] In another embodiment, the bacteriophage used in the present invention can be isolated from any environment in which bacteria exist, hi some embodiments, the recombinant bacteriophage of the present invention are isolated (e.g., harvested, obtained) from fecal or sewage, terrestrial, or marine environments.

[0082] Treatment method Also provided herein is a method for preventing, treating, and / or reducing the risk of cancer, autoimmune diseases, respiratory diseases (e.g., asthma), neurological diseases, infectious diseases, and other diseases (osteoporosis, hypercholesterolemia, etc.), which comprises administering to a subject in need thereof a probiotic vaccine of the present invention containing a heterologous peptide selected from SEQ ID NOs: 9 to 133 described herein and / or a recombinant bacteriophage of the present invention.

[0083] In certain embodiments, there is provided a method for preventing, treating, and / or reducing the risk of cancer, autoimmune diseases, respiratory diseases (e.g., asthma), neurological diseases, infectious diseases, and other diseases (osteoporosis, hypercholesterolemia, etc.), comprising administering to a subject in need thereof a probiotic vaccine of the present invention comprising a heterologous peptide selected from SEQ ID NOs: 9-133 described herein. In another embodiment, there is provided a method for preventing, treating, and / or reducing the risk of cancer, autoimmune diseases, respiratory diseases (e.g., asthma), neurological diseases, infectious diseases, and other diseases (osteoporosis, hypercholesterolemia, etc.), comprising administering to a subject in need thereof a recombinant bacteriophage of the present invention comprising a heterologous peptide selected from SEQ ID NOs: 9-133 described herein.

[0084] Also provided herein are methods for preventing, treating, and / or reducing the risk of cancer, autoimmune diseases, respiratory diseases (e.g., asthma), neurological diseases, infectious diseases, and other diseases (e.g., osteoporosis, hypercholesterolemia), comprising administering to a subject in need thereof a probiotic vaccine of the present invention in combination with a recombinant bacteriophage of the present invention comprising a heterologous peptide selected from SEQ ID NOS: 9-133 described herein. In certain embodiments, probiotic vaccines of the present invention comprising recombinant bacteriophages of the present invention have been shown to have the ability to penetrate mucosal layers and generate significant secretory IgA (S1gA) immune responses, enabling the body to respond to intramucosal antigens and malignancies (see Figure 3 and Example 10). In another embodiment, recombinant bacteriophages of the present invention (e.g., filamentous phage) have been shown to have the ability to generate significant IgA and IgG immune responses when administered by injection and / or intravenously (IV) (see Figures 2A and 2B and Example 9). Therefore, also provided herein are therapeutic methods comprising administering in combination the probiotic vaccine of the present invention (e.g., oral administration, etc.) and the recombinant bacteriophage of the present invention (e.g., injection, etc.).

[0085] cancer As used herein, the term "cancer," or grammatical variations thereof, refers to a malignant tumor. In particular, the term "cancer" refers to any of a group of diseases or disorders characterized by uncontrolled cell division and the ability of those cells to invade other tissues, either by direct growth into adjacent tissues via invasion, or by transplantation to distant sites via metastasis. Metastasis is defined as the stage at which cancer cells migrate through the bloodstream or lymphatic system. In certain embodiments, cancer is Multiple myeloma, epithelial carcinoma, epithelial ovarian cancer, mucosal melanoma, non-small cell lung cancer, melanoma, head and neck cancer, renal cell carcinoma, Hodgkin's lymphoma, cutaneous squamous cell carcinoma, glioblastoma, esophageal cancer, gastric cancer, duodenal cancer, small intestine cancer, appendix cancer, colon cancer, rectal cancer, colorectal cancer, anal cancer, pancreatic cancer, liver cancer, gallbladder cancer, spleen cancer, kidney cancer, bladder cancer, prostate cancer, testicular cancer, uterine cancer, endometrial cancer, ovarian cancer, vaginal cancer, vulvar cancer, breast cancer, lung cancer, thyroid cancer, thymus cancer, brain cancer, nervous system cancer, glioblastoma In certain embodiments, the cancer is selected from the group consisting of multiple myeloma, oral cancer, skin cancer, blood cancer, lymphoma, eye cancer, bone cancer, bone marrow cancer, muscle cancer, non-small cell lung cancer (NSCLC), head and neck squamous cell carcinoma (HNSCC), urothelial cancer, non-muscle invasive bladder cancer (NMIBC), colon or rectal cancer, esophageal cancer or certain gastroesophageal junction (GEJ) cancers, cervical cancer, renal cell carcinoma (RCC), advanced endometrial cancer, cutaneous squamous cell carcinoma (cSCC), and / or triple-negative breast cancer (TNBC), etc. In certain embodiments, the cancer is multiple myeloma.

[0086] In certain embodiments, as described above, provided herein are methods for preventing, treating, and / or reducing the risk of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, and / or gastric cancer, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from NPLVA (SEQ ID NO: 11), NPLVANPLVA (SEQ ID NO: 46), and / or FLAEDALNTV (SEQ ID NO: 126), or a fragment or variant thereof.

[0087] In certain embodiments, recombinant phage and / or probiotic vaccines comprising FLAEDALNTV (SEQ ID NO: 126) are used to prevent, treat or reduce the risk of breast cancer.

[0088] In another embodiment, as described above, provided herein is a method of preventing, treating and / or reducing the risk of multiple myeloma, comprising administering to a subject in need thereof: KRNIQFSCKNI (SEQ ID NO: 49), QTLEAQTLEA (SEQ ID NO: 58), and / or IQFSCIQFSC (SEQ ID NO: 59) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of:

[0089] In another embodiment of the use of PD-1 exogenous peptide epitopes (or PD-1-associated epitopes) for the prevention, treatment, and / or risk reduction of cancer, at least one exogenous peptide epitope corresponding to ESFDGDPASNTAPLQPEQL (SEQ ID NO: 38), APKAQIKESLR (SEQ ID NO: 64), EDPAFYMLKGKLQYELQ (SEQ ID NO: 84), and / or SPRRKLISVDSRSVSL (SEQ ID NO: 131), or a fragment thereof. Alternatively, the variants may prevent, treat and / or reduce the risk of PD-1 mediated cancers such as, for example, mucosal cancer, gastric cancer, melanoma, non-small cell lung cancer (NSCLC), head and neck squamous cell carcinoma (HNSCC), urothelial carcinoma, non-muscle invasive bladder cancer (NMIBC), colon or rectal cancer, esophageal cancer or certain gastroesophageal junction (GEJ) cancers, cervical cancer, renal cell carcinoma (RCC), advanced endometrial cancer, cutaneous squamous cell carcinoma (cSCC) and / or triple-negative breast cancer (TNBC). In another embodiment, the exogenous peptide epitope comprises ESFDGDPASNTAPLQPEQL (SEQ ID NO: 38), APKAQIKESLR (SEQ ID NO: 64), EDPAFYMLKGKLQYELQ (SEQ ID NO: 84) and / or SPRRKLISVDSRSVSL (SEQ ID NO: 131), with no more than 5, no more than 4, no more than 3, no more than 2, or no more than 1 amino acid substitution within each peptide epitope.Accordingly, provided herein is a method of preventing, treating and / or reducing the risk of cancer, comprising administering to a subject in need thereof a recombinant phage and / or a recombinant phage comprising at least one exogenous peptide epitope corresponding to ESFDGDPASNTAPLQPEQL (SEQ ID NO: 38), APKAQIKESLR (SEQ ID NO: 64), EDPAFYMLKGKLQYELQ (SEQ ID NO: 84) and / or SPRRKLISVDSRSVSL (SEQ ID NO: 131), or a fragment or variant thereof. and / or a probiotic vaccine, wherein the cancer is selected from the group consisting of gastric cancer, melanoma, non-small cell lung cancer (NSCLC), head and neck squamous cell carcinoma (HNSCC), urothelial carcinoma, non-muscle-invasive bladder cancer (NMIBC), colon or rectal cancer, esophageal cancer or certain gastroesophageal junction (GEJ) cancers, cervical cancer, renal cell carcinoma (RCC), advanced endometrial cancer, cutaneous squamous cell carcinoma (cSCC), and / or triple-negative breast cancer (TNBC).

[0090] In certain embodiments, as described above, provided herein are methods for preventing, treating and / or reducing the risk of mucosal melanoma, comprising administering to a subject in need thereof; ALLEIASCL (SEQ ID NO: 9), EDPAFYMLKGKLQYELQ (SEQ ID NO: 84), MYPPPY (SEQ ID NO: 10), MYPPPYMYPPPY (SEQ ID NO: 43), VELMYPPPYYLGIG (SEQ ID NO: 44), LMYPPPYYLGIGN (SEQ ID NO: 45), and / or SPRRKLISVDSRSVSL (SEQ ID NO: 131) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of:

[0091] In certain embodiments, as described above, provided herein are methods of preventing, treating and / or reducing the risk of prostate cancer, comprising administering to a subject in need thereof: ESKVDPSKAW (SEQ ID NO: 29), ESKVDPSK (SEQ ID NO: 12), MYPPPY (SEQ ID NO: 10), MYPPPYMYPPPY (SEQ ID NO: 43), VELMYPPPYYLGIG (SEQ ID NO: 44), LMYPPPYYLGIGN (SEQ ID NO: 45), ASPLLSGSQHFDVPPRQL (SEQ ID NO: 65), and / or KVIQAQTAFSANPANPAILS (SEQ ID NO: 67) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of:

[0092] In certain embodiments, as described above, provided herein are methods for preventing, treating, and / or reducing the risk of hematological cancer, leukemia, chronic lymphocytic leukemia (CLL), small lymphocytic lymphoma (SLL), Waldenstrom's macroglobulinemia, chronic graft-versus-host disease, mantle cell lymphoma (MCL), marginal zone lymphoma, and the like, comprising administering to a subject in need thereof: PTPEEDQILKK (SEQ ID NO: 63), DVIGRIENGERL (SEQ ID NO: 119), and / or RLIKEDVYL (SEQ ID NO: 120) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of:

[0093] In certain embodiments, as described above, provided herein are methods for preventing, treating, and / or reducing the risk of acute myeloid leukemia (AML), autoimmune and inflammatory diseases, canine cancer, and the like, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope corresponding to ASPLLSGSQHFDVPPRQL (SEQ ID NO: 65) or a fragment or variant thereof.

[0094] In certain embodiments, as described above, provided herein are methods for preventing, treating, and / or reducing the risk of pulmonary fibrosis, comprising administering to a subject in need thereof: QYDSWPHYPE (SEQ ID NO: 66), YLSETVSNALGPQGRRLEE (SEQ ID NO: 73), EDPAFYMLKGKLQYELQ (SEQ ID NO: 84), MYPPPY (SEQ ID NO: 10), MYPPPYMYPPPY (SEQ ID NO: 43), VELMYPPPYYLGIG (SEQ ID NO: 44) LMYPPPYYLGIGN (SEQ ID NO: 45), and / or SPRRKLISVDSRSVSL (SEQ ID NO: 131) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of:

[0095] In certain embodiments, as described above, provided herein are methods for preventing, treating, and / or reducing the risk of lung cancer, non-small cell lung cancer, etc., comprising administering to a subject in need thereof: SPRMSGLLSQTGYLSDNHILI (SEQ ID NO: 75), ASPLLSGSQHFDVPPRQL (SEQ ID NO: 65), SSDWVTLGTSASLLR (SEQ ID NO: 90), STPYVGLSLSCAANQ (SEQ ID NO: 91), PSPQYSWRINGIPQQHTQVLF (SEQ ID NO: 92), NPTTYQMDVNPEGKYS (SEQ ID NO: 95), HTPPLDPQELDILKT (SEQ ID NO: 96), RPLKGAPLPAL (SEQ ID NO: 101), EDPAFYMLKGKLQYELQ (SEQ ID NO: 84), MYPPPY (SEQ ID NO: 10), VVVGADVGKS (SEQ ID NO: 62), KRAS, VVVGACVGKS (SEQ ID NO: 129), KRAS, MYPPPYMYPPPY (SEQ ID NO: 43), VELMYPPPYYLGIG (SEQ ID NO: 44), LMYPPPYYLGIGN (SEQ ID NO: 45), GRADALRVLSK (SEQ ID NO: 122), LSRELSSLASQPLSR (SEQ ID NO: 123), and / or SPRRKLISVDSRSVSL (SEQ ID NO: 131) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of:

[0096] In certain embodiments, as described above, provided herein are methods for preventing, treating, and / or reducing the risk of thyroid eye disease, etc., comprising administering to a subject in need thereof: GRADALRVLSK (SEQ ID NO: 122), and / or LSRELSSLASQPLSR (SEQ ID NO: 123) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of:

[0097] Provided herein are methods for preventing, treating, and / or reducing the risk of cancer, comprising administering to a subject in need thereof: PIPHDGNLYPRLYPE (SEQ ID NO: 68), SKCRRCRLLD (SEQ ID NO: 69), QNTKCRCKPNFFC (SEQ ID NO: 70), TETEPPVAL (SEQ ID NO: 117), REALHEAL (SEQ ID NO: 118), VVVGADVGKS (SEQ ID NO: 62), KRAS, VVVGACVGKS (SEQ ID NO: 129), KRAS, and / or KTLETNPYVK (SEQ ID NO: 121) administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope, or a fragment or variant thereof, selected from the group consisting of: The methods are provided wherein the cancer is selected from the group consisting of gastric cancer, melanoma, mucosal melanoma, lung cancer, non-small cell lung cancer (NSCLC), head and neck squamous cell carcinoma (HNSCC), urothelial carcinoma, non-muscle-invasive bladder cancer (NMIBC), colon or rectal cancer, esophageal cancer or certain gastroesophageal junction (GEJ) cancers, cervical cancer, renal cell carcinoma (RCC), advanced endometrial cancer, cutaneous squamous cell carcinoma (cSCC), pancreatic ductal carcinoma, and / or prostate cancer.

[0098] In certain embodiments, recombinant phage and / or probiotic vaccines comprising KTLETNPYVK (SEQ ID NO: 121) are used to prevent, treat or reduce the risk of colon cancer.

[0099] infectious disease In another embodiment, as described above, provided herein is a method for preventing Korean fever virus infection and / or disease manifestations, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine of the present invention comprising at least one exogenous peptide epitope selected from KRNIQFSCKNI (SEQ ID NO: 49), QTLEAQTLEA (SEQ ID NO: 58) and / or IQFSCIQFSC (SEQ ID NO: 59), or a fragment or variant thereof.

[0100] In another embodiment, provided herein is a method of preventing Covid infection (e.g., Covid-19 infection) and / or disease manifestations, comprising administering to a subject in need thereof: AGICAS (SEQ ID NO: 14), AG-I-CAS (SEQ ID NO: 15; Ile = "AUA" codon), VLYQDVNCT (SEQ ID NO: 19) QQQQGQTITKKSAAEASKKPRQKRTAVLVQDVGCTFSTVFPVTSFGPLVR (SEQ ID NO: 20), VLYQGVNCT (SEQ ID NO: 21), AGICASA GICAS (SEQ ID NO: 28), CVADYSVLY (SEQ ID NO: 33), HVGEIPVAY (SEQ ID NO: 34), NQKLIANQF (NQK-Q8) (SEQ ID NO: 35), NQKLIANAF (NQK-A8) (SEQ ID NO: 36), and / or RVAGDSGFAAY (SEQ ID NO: 37) The method comprises administering a recombinant phage and / or probiotic vaccine of the present invention comprising at least one exogenous peptide epitope selected from the group consisting of:

[0101] In a specific embodiment, the probiotic vaccine of the present invention utilizes the AGICAS (SEQ ID NO: 14) exogenous peptide epitope, in which the isoleucine is encoded by the "AUA" (or ATA) codon, referred to herein as SEQ ID NO: 15. The AGICAS epitope is within the COVID-19 spike protein, and the related AGFCAS peptide is within platelet factor 4. This molecular mimicry is responsible for some of the cardiac problems associated with current vaccines, as the isoleucine "AUU" codon in the AGICAS peptide epitope is inadvertently mutated to the phenylalanine "UUU" codon, causing the vaccine to produce antibodies against the AGFCAS epitope in platelet factor 4, which can also lead to blood clots. According to the present invention, the codon used for Ile is "AUA," and two mutations are required to result in the "UUU" codon for PHE. By using this particular "AUA" codon, the likelihood of harmful PHE mutations is reduced by a factor of two. It is contemplated herein that use of the specific codon "AUU" for Ile reduces the probability of thrombosis from 1 in 300,000 to about 1 in 10 billion. It is also contemplated herein to use two or more copies of the AGICAS (SEQ ID NO: 14) peptide epitope, for example, 2 copies, 3 copies, 4 copies, 5 copies, 6 copies, 7 copies, 8 copies, 9 copies, 10 copies, or more copies. In a specific embodiment, two copies corresponding to AGICASAGICAS (SEQ ID NO: 28) are used herein.

[0102] Thus, in a particular embodiment, the isoleucine (Ile) in the AGICAS SEQ ID NO: 14 sequence is specifically encoded by the "AUA" codon. Similarly, in another embodiment, one or both of the Ile amino acids in the AGICASA GICAS SEQ ID NO: 28 sequence are specifically encoded by the "AUA" codon.

[0103] In another embodiment, provided herein is a method for preventing monkeypoxvirus infection and / or disease development, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine of the present invention comprising at least one exogenous peptide epitope selected from AKPTAKREAIVKAYG (SEQ ID NO: 16), ISPDGCYSL (SEQ ID NO: 17), LTFDYVVTF (SEQ ID NO: 18), LTFDY (SEQ ID NO: 50) and / or LTFDYLTFDY (SEQ ID NO: 51), or a fragment or variant thereof.

[0104] In certain embodiments, as described above, provided herein are methods for preventing, treating, and / or reducing the risk of yellow fever, the methods comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope corresponding to LTHVKIND (SEQ ID NO: 55) or a fragment or variant thereof.

[0105] In certain embodiments, as described above, provided herein are methods for preventing, treating and / or reducing the risk of malaria, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope corresponding to NANPNANPNANPNAN (SEQ ID NO: 56), or a fragment or variant thereof.

[0106] In certain embodiments, as described above, provided herein are methods for preventing, treating and / or reducing the risk of tuberculosis, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope corresponding to MTEQQWNFAGIEAAASAIQG (SEQ ID NO: 102), or a fragment or variant thereof.

[0107] In certain embodiments, as described above, provided herein is a method for preventing, treating and / or reducing the risk of leprosy, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from KTIAYDEEARR (SEQ ID NO: 103) and / or EEFVVEFDLPGIKA (SEQ ID NO: 104), or a fragment or variant thereof.

[0108] In certain embodiments, as described above, provided herein are methods for preventing, treating, and / or reducing the risk of poliovirus infection and / or disease manifestations, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope corresponding to CVTIMTVDNPASTTNKDKLFAVWKITYKDT (SEQ ID NO: 105), or a fragment or variant thereof.

[0109] In certain embodiments, as described above, provided herein are methods for preventing, treating, and / or reducing the risk of human immunodeficiency virus (HIV) infection and / or disease manifestations, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from ILKEPVHGV (SEQ ID NO: 106) and / or SLYNTVATL (SEQ ID NO: 107), or a fragment or variant thereof.

[0110] In certain embodiments, as described above, provided herein are methods for preventing, treating, and / or reducing the risk of measles, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope corresponding to LSEIKGVIVHRLEGV (SEQ ID NO: 108), or a fragment or variant thereof.

[0111] In certain embodiments, as described above, the present specification provides a method for preventing, treating, and / or reducing the risk of plague infection (Yersinia Pestis), comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from MLMAASRAL (SEQ ID NO: 109), RAMRMVYYL (SEQ ID NO: 110), and / or FFVRSIGSKGGKLAAGKYTDAVTV (SEQ ID NO: 111), or a fragment or variant thereof.

[0112] In certain embodiments, as described above, provided herein are methods for preventing, treating, and / or reducing the risk of Leishmania chagasi, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope corresponding to EEPEESDEDDFGMGGLF (SEQ ID NO: 112), or a fragment or variant thereof.

[0113] In certain embodiments, as described above, provided herein are methods for preventing, treating and / or reducing the risk of hepatitis C, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from CINGVCWTV (SEQ ID NO: 113) and / or KLVALGINAV (SEQ ID NO: 114), or a fragment or variant thereof.

[0114] In certain embodiments, as described above, provided herein are methods for preventing, treating, and / or reducing the risk of shingles, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from NLVPMVATV (SEQ ID NO: 115) and / or TPRVTGGGAM (SEQ ID NO: 116), or a fragment or variant thereof.

[0115] In certain embodiments, as described above, provided herein are methods for preventing, treating, and / or reducing the risk of Alaskapox, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from QEANASAQTKCDIE (SEQ ID NO: 128), or a fragment or variant thereof.

[0116] In certain embodiments, as described above, provided herein are methods for preventing, treating and / or reducing the risk of dengue virus infection and / or disease manifestations, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from LPAIVREAI (SEQ ID NO: 130), or a fragment or variant thereof.

[0117] In certain embodiments, as described above, provided herein are methods for preventing, treating, and / or reducing the risk of developing Bordetella pertussis infection and / or pertussis disease, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from DNVLDHLTGRSC (SEQ ID NO: 133), or a fragment or variant thereof.

[0118] In another embodiment, provided herein is a method for preventing, treating and / or reducing the risk of avian influenza infection and / or disease manifestations, comprising administering to a subject in need thereof: DAPF (SEQ ID NO: 22), KQIVERILEEE (SEQ ID NO: 23), RYLTDMTLEE (SEQ ID NO: 24), DDVKRENPEHEEYNN (SEQ ID NO: 25), RESGSS (SEQ ID NO: 26), DAPFKQIVERILEEE (SEQ ID NO: 30), RESGSSRYLTDMTLEE (SEQ ID NO: 31), EFINVPEGGS (SEQ ID NO: 39), EFINVPEEFINVPE (SEQ ID NO: 40), DAPFDAPF (SEQ ID NO: 60), and / or RESGSSRESGSS (SEQ ID NO: 61) The method comprises administering a recombinant phage and / or probiotic vaccine of the present invention comprising at least one exogenous peptide epitope selected from the group consisting of:

[0119] immunotherapy Immunotherapy (immuno-oncology therapy) refers to the induction, promotion and / or modulation of a therapeutic immune response by inducing the production of antibodies against a specific target, such as TNF. Thus, immunotherapy refers to the engineering or generation of a therapeutic immune response by inducing the production of antibodies against a specific target.

[0120] In certain embodiments, as described above, provided herein are methods for preventing, treating, and / or reducing the risk of rheumatoid arthritis, juvenile idiopathic arthritis, psoriatic arthritis, ankylosing spondylitis, Crohn's disease, ulcerative colitis, plaque psoriasis, hidradenitis suppurativa, uveitis, and the like, comprising administering to a subject in need thereof: RSSSRTPS (SEQ ID NO: 13), VRSSSRTPSD (SEQ ID NO: 41), RSSSRTPSDK (SEQ ID NO: 42), ETPEGAEA (SEQ ID NO: 48), and / or ALDENTKLWYA (SEQ ID NO: 87) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of:

[0121] In certain embodiments, as described above, provided herein are methods for preventing, treating and / or reducing the risk of multiple sclerosis, comprising administering to a subject in need thereof: ESVDPKNYPKK (SEQ ID NO: 72), RPKSNIVLLS (SEQ ID NO: 97), RPKSNIVLLSAEEKKEQT (SEQ ID NO: 99), and / or SGPKPLFRRMSSLVGPTQ (SEQ ID NO: 98) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of:

[0122] In certain embodiments, as described above, provided herein are methods for preventing, treating, and / or reducing the risk of osteoporosis, sclerosis, Van Buchem disease, osteonecrosis, osteoarthritis, rheumatoid arthritis, ankylosing spondylitis, bone tumors, multiple myeloma, bone metastasis, prostate cancer, glucocorticoid-induced osteoporosis, etc., comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope corresponding to LPNAIGRGKWWRP (SEQ ID NO: 47) and / or CTHYMQLAC (SEQ ID NO: 127), or a fragment or variant thereof.

[0123] In certain embodiments, as described above, provided herein are methods for preventing, treating, and / or reducing the risk of primary hyperlipidemia, heterozygous familial hypercholesterolemia, hyperlipidemia, established cardiovascular disease, such as reducing the risk of myocardial infarction, stroke, and coronary revascularization, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope corresponding to REIEGRVMVTDPENVPEEDG (SEQ ID NO: 81), or a fragment or variant thereof.

[0124] In certain embodiments, as described above, provided herein are methods for preventing, treating, and / or reducing the risk of acne, signs of aging, wrinkles, fine lines, scars, hyperpigmentation, Kaposi's sarcoma lesions, melasma, psoriasis, stretch marks, and the like, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope corresponding to GYYYRDDSLPVLLE (SEQ ID NO: 83), or a fragment or variant thereof.

[0125] In certain embodiments, as described above, provided herein are methods for preventing, treating, and / or reducing the risk of paroxysmal nocturnal hemoglobinuria, atypical hemolytic uremic syndrome, intravascular hemolysis, thrombosis, fatigue, the need for blood transfusions, and the like, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope corresponding to DKPVYTPDQSVKVRV (SEQ ID NO: 132), or a fragment or variant thereof.

[0126] In certain embodiments, as described above, provided herein are methods for preventing, treating, and / or reducing the risk of asthma, severe asthma, atopic dermatitis, eczema, eosinophilic or oral steroid-dependent asthma, chronic rhinosinusitis with nasal polyposis, eosinophilic esophagitis, prurigo nodularis, and the like, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope corresponding to NTVSCNRPHCITE (SEQ ID NO: 52), EERRR (SEQ ID NO: 53), EERRREERRR (SEQ ID NO: 54), and / or AGAFAPSAAVA (SEQ ID NO: 82), or a fragment or variant thereof.

[0127] In certain embodiments, as described above, provided herein is a method for preventing, treating, and / or reducing the risk of asthma or severe eosinophilic asthma, etc., comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope corresponding to (SEQ ID NO: 52), EERRR (SEQ ID NO: 53), and / or EERRREERRR (SEQ ID NO: 54), or a fragment or variant thereof.

[0128] In certain embodiments, as described above, provided herein are methods for preventing, treating, and / or reducing the risk of psoriasis, comprising administering to a subject in need thereof: LCTLAWAHPLVGHM (SEQ ID NO: 71), ALFDAQAQV (SEQ ID NO: 85), LLYDLTEKQVY (SEQ ID NO: 86), APNRTITVDDKMSLRLH (SEQ ID NO: 89), SLLPDSPVGQ (SEQ ID NO: 124), and / or SPSQPWQRL (SEQ ID NO: 125) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of:

[0129] In another embodiment of the present invention, provided herein is a method of preventing, treating and / or reducing the risk of an inflammatory disease selected from the group consisting of psoriatic arthritis, Crohn's disease, ulcerative colitis, psoriasis, rheumatoid arthritis, atopic dermatitis, ankylosing spondylitis, hidradenitis suppurativa, uveitis, and the like, comprising administering to a subject in need thereof: EIRQAGRPNKPDSI (SEQ ID NO: 57), and / or VLVESEHQV (SEQ ID NO: 88) The method comprises administering a recombinant phage or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of:

[0130] Neurological disorders In certain embodiments, as described above, provided herein are methods for preventing, treating, and / or reducing the risk of pain, comprising administering to a subject in need thereof: GLPSLGLTL (SEQ ID NO: 76), HRGEFS (SEQ ID NO: 77), HRGEFSHRGEFS (SEQ ID NO: 78), STLYREELMTTLLYYEIFL (SEQ ID NO: 93), and / or CEKNSVLEVIAYSSSE (SEQ ID NO: 94) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of:

[0131] In certain embodiments, as described above, provided herein are methods for preventing, treating and / or reducing the risk of chronic wasting, mad cow disease, etc., comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope corresponding to DYEDRYYRE (SEQ ID NO: 100), or a fragment or variant thereof.

[0132] In another embodiment, provided herein is a method of preventing, treating and / or reducing the risk of a neurological disorder, such as Alzheimer's disease, Parkinson's disease, dementia, multiple sclerosis, epilepsy, Huntington's disease, muscular dystrophy, or migraine, comprising administering to a subject in need thereof: FRHDSGY (SEQ ID NO: 27), THLHWHTV (SEQ ID NO: 79), VQDYVQMKASELEQ (SEQ ID NO: 80), and / or INDPRLRNPSDKFIY (SEQ ID NO: 74) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of: In a specific embodiment, the disease to be treated or prevented is Alzheimer's disease. In another embodiment, the disease to be treated or prevented with a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from VQDYVQMKASELEQ (SEQ ID NO: 80), or a fragment or variant thereof, is migraine.

[0133] Administration of probiotic vaccines The probiotic vaccines (e.g., bacterial cells containing the recombinant phage of the invention) and / or recombinant bacteriophage pharmaceutical compositions of the invention can be delivered or administered to a subject (e.g., a patient suffering from cancer) by any suitable delivery route, including, but not limited to, pulmonary, intramuscular, subcutaneous, intubated (intratracheal), or intraperitoneal routes. The route of administration will be determined by the attending medical professional to be the most appropriate route taking into account all factors related to the subject. The subject may be any suitable subject, including, but not limited to, humans, primates, dogs, cats, horses, cattle, etc. In one embodiment, the subject is a human.

[0134] In some embodiments, live cells of probiotic bacteria and / or recombinant bacteriophages can be orally administered as probiotic vaccines of the present invention via fermented dairy products and / or probiotic-fortified foods. Examples of delivery foods include probiotic solutions, gummies (e.g., gelatin-based), tablets, yogurt, coconut water, pickles, fermented bean pastes (e.g., tempeh, miso, doenjang), kefir, buttermilk or carne melk, kimchi, pao cai, sauerkraut, soy sauce, and pickled mustard greens. In another embodiment, probiotic bacteria can be delivered to the body as tablets, capsules, powders, and / or sachets containing the bacteria in lyophilized form.

[0135] In certain embodiments, the pharmaceutical carrier is a gelatin-based chewable formulation (e.g., a gummy or lozenge). In this embodiment, the gummies of the present invention are translucent, tasteless, or odorless, and comprise an anhydrous gelatin gummies base derived from naturally occurring ingredients, which facilitates the formulation of water-labile APIs or can be used when the stability of the drug is unknown. In this embodiment, the gummies base is heated, mixed with other ingredients, poured into a mold, and cooled. In one embodiment, the base has a relatively low working temperature when heated, forming a pourable solution, and upon cooling, forms a stable gelatin matrix with the desired API loading capacity. The gummies of the present invention can be easily sweetened and flavored to suit individual preferences.

[0136] In certain embodiments, commercially available KlearGummy™ (SpecializedRx Products, Minnesota) is used, which provides a ready-to-use, anhydrous gummy base that meets USP standards for extended shelf life (BUD) stability profile. This base has a slight translucent amber color and can be loaded with active ingredients, sweeteners, and flavorings of choice.

[0137] In another embodiment, the probiotic vaccine and / or recombinant bacteriophage of the present invention can be delivered to a subject by injection, infusion, inoculation, direct surgical delivery, or any combination thereof. In some embodiments, the probiotic vaccine and / or recombinant bacteriophage is administered to a human in the deltoid region or axillary region. For example, the probiotic vaccine and / or recombinant bacteriophage is administered as an intradermal injection in the axillary region. In another embodiment, the probiotic vaccine and / or recombinant bacteriophage is administered intravenously by injection or infusion.

[0138] In certain embodiments, incorporating the recombinant bacteriophages of the present invention into therapeutic formulations involves encapsulating them with a stabilizing material. This method allows for the production of a variety of antimicrobial materials, such as powders, semi-solids, and nanofibers, providing a wide range of options for effective delivery. Thus, various encapsulated phage formulations are contemplated in the present invention, including spray powders, lyophilized powders, emulsions, and liposomes. For further details, see Bulbake et al., "Antimicrobial Agents for Antimicrobial Agents," vol. 1, no. 1, pp. 111-114, 2002, which is incorporated herein by reference in its entirety for all purposes. Liposomal formulations in clinical use: An updated review . Pharmaceutics. 2017;9:12. doi: 10.3390 / pharmaceutics9020012;Esteban et al., Enhancement of the antimicrobial properties of bacteriophage-k via stabilization using oil-in-water nano-emulsions . Biotechnol. Prog. 2014;30:932-944. doi: 10.1002 / btpr.1898.

[0139] Also contemplated herein as therapeutic formulations of the recombinant phages of the present invention are immobilization methods in which the bacteriophage is coated / surrounded by specific stabilizing agents to provide protection from the external environment, in which the phage is bound to a substrate surface, and / or encapsulation methods using any of emulsification, lyophilization, spray drying, liposome encapsulation, and electrospinning, as described in Rosner et al., Pharmaceuticals (Basel). 2021 Apr; 14(4): 359; Published online 2021 Apr 13. doi: 10.3390 / ph14040359, the entire contents of which are incorporated herein by reference for all purposes.

[0140] In another embodiment, the compositions of the invention (e.g., recombinant bacteriophages or probiotic vaccines) are administered pulmonary. For example, freeze-drying and spray-drying methods can be used to create phage-coated powders, which can be used in creams for direct application, tablets for oral administration, or even incorporated into inhaler systems. See, for example, Malik et al., 2004, Pharmacology of Microbial Diseases, Vol. 1, No. 1, pp. 111-114, 2004, which is incorporated herein by reference in its entirety for all purposes. Formulation, stabilization and encapsulation of bacteriophage for phage therapySee Adv. Colloid Interface Sci. 2017;249:100-133. doi: 10.1016 / j.cis.2017.05.014. In certain embodiments of the invention, pulmonary administration comprises inhalation of a therapeutic dose (e.g., single dose or multiple doses) of a recombinant phage pharmaceutical composition of the invention described herein via nasal inhalation, oral inhalation, or both. The recombinant phage pharmaceutical composition can be administered in two or more separate administrations (doses). In one embodiment, the recombinant phage can be formulated as a dry powder from a dry powder inhaler (either alone, in a mixture, or in a dry blend, for example, with lactose) or as an aerosol spray (e.g., droplets of a stable dispersion or suspension of a probiotic vaccine in a gaseous medium). See, for example, Chang et al., Antimicrob. Agents Chemother. 2018;62:e01714-17. doi: 10.1128 / AAC.01714-17, the entirety of which is incorporated by reference herein for all purposes. Recombinant phage compositions of the present invention delivered by aerosol may be deposited in the respiratory tract by gravitational settling, inertial impaction, and / or diffusion. Any suitable device for generating an aerosol can be used, including, but not limited to, pressurized MDIs, nebulizers, vaporizers (inhalation via vaping devices, e-cigarettes), dry powder inhalers (DPIs), and soft mist inhalers.

[0141] In certain embodiments, the recombinant phage of the present invention is nebulized as described in Guellec et al., Viruses. 2023 Mar; 15(3): 602 (doi: 10.3390 / v15030602), the entire contents of which are incorporated herein by reference for all purposes. It is believed herein that the ability of the phage to self-replicate in the presence of the target may reduce dosage and improve compliance. The specificity of the recombinant phage of the present invention limits its impact on the microflora involved in maintaining tissue homeostasis. Phages are generally considered safe, can be administered via various routes, and have a low incidence of adverse events.

[0142] In another embodiment, a suitable carrier for administering cells can be routinely selected by those skilled in the art. For example, pharmaceutical carriers include buffered saline, such as cell culture medium (e.g., LB medium), and in another embodiment, DMSO to maintain cell viability.

[0143] In one embodiment, the probiotic vaccine and / or recombinant bacteriophage pharmaceutical composition of the present invention comprises a suspension further comprising a pharmaceutically acceptable aqueous carrier. The suspension can comprise a probiotic vaccine bacterium and / or recombinant phage and a liquid carrier. The liquid carrier can be aqueous. The liquid carrier of the suspension can comprise water, coconut water, and / or LB medium, and optionally one or more excipients selected from the group consisting of a buffer, a tonicity adjuster, a preservative, a demulcent, a viscosity adjuster, an osmotic agent, a surfactant, an antioxidant, an alkalizing agent, an acidifying agent, an antifoaming agent, and a colorant. For example, in one embodiment, the suspension can comprise a probiotic vaccine bacterium of the present invention, coconut water, a buffer, and a salt. Optionally, it can further comprise a surfactant. In some embodiments, the suspension consists essentially of or consists of coconut water, a suspension of the probiotic vaccine bacterium of the present invention and / or a recombinant phage of the present invention in coconut water and a buffer. The suspension can further comprise an osmotic salt.

[0144] In certain embodiments, the pharmaceutical carrier is coconut water. In one embodiment, the probiotic vaccine bacteria infected with the recombinant phage of the present invention can remain in the coconut water medium used to culture the respective probiotic vaccine bacteria and then be administered directly to a subject. In another embodiment, the probiotic vaccine bacteria infected with the recombinant phage of the present invention can be cultured in a separate medium and then introduced into coconut water as a final formulation and then be administered directly to a subject.

[0145] In another embodiment, it is also contemplated herein that the probiotic vaccine bacteria infected with the recombinant phage of the present invention can be left in the LB medium used to grow the respective probiotic vaccine bacteria and can be administered directly to a subject.

[0146] In certain embodiments, the cells are administered in an injectable cryopreservation medium, the composition comprising the cells including DMSO and hetastarch, Plasmalyte A and / or a glucose solution as cryoprotectants, and human serum albumin as a protein component.

[0147] The amount of probiotic vaccine and / or recombinant bacteriophage administered to a subject as a cancer vaccine to practice the methods described herein, and the most convenient route of such administration, as well as the formulation of the vaccine itself, are determined based on a variety of factors. These factors include the subject's physical characteristics (e.g., age, weight, sex), the physical characteristics of the tumor (e.g., location, size, growth rate, accessibility), and the extent of other treatments (e.g., chemotherapy, radiation therapy) administered in conjunction with the overall treatment plan. Despite the various factors that must be considered when practicing the methods of the present invention for preventing, treating, and / or reducing the risk of developing cancer or an infectious disease, a single dose of about 5 x 10 virions in about 0.05 mL to about 2 mL of solution (e.g., saline) is generally administered to mammals, preferably humans. 6 ~about 2×10 8 In another embodiment, a mammal, preferably a human, can be administered about 1 x 10 probiotic vaccine cells in about 0.05 mL to about 2 mL of solution (e.g., saline) in a single dose. 3 ~Approx. 1×10 15 , 1×10 4 ~Approx. 1×10 14 , 1×10 5 ~Approx. 1×10 13 , 1×10 5 ~Approx. 1×10 12 , 1×10 5 ~Approx. 1×10 11 , 1×10 5 ~Approx. 1×10 10 , 1×10 5 ~Approx. 1×10 9 , 1×10 5 ~Approx. 1×10 8 , 1×10 6 ~Approx. 1×10 12 , 1×10 6 ~Approx. 1×10 11 , 1×10 6 ~Approx. 1×10 10 , 1×10 6 ~Approx. 1×10 9 , 1×10 6 ~Approx. 1×10 8 probiotic vaccine cells are administered.

[0148] Additional doses may be administered depending on the aforementioned factors, such as the severity of the tumor lesion, and other factors. In a further embodiment, about 1 x 10 4 / ml, approx. 1×10 5 / ml, approx. 1×10 6 / ml, approx. 1×10 7 / ml, approx. 1×10 8 / ml, approx. 1×10 9 / ml, approx. 1×10 10 / ml, approx. 1×10 11 / ml, approx. 1×10 12 / ml, approx. 1×10 13 / ml, approx. 1×10 14 / ml, approx. 1×10 15 / ml of probiotic vaccine cells is administered about 1 to about 5 times at 2-week intervals. 8 / ml of probiotic vaccine cells is administered about 1 to about 5 times at 2-week intervals.

[0149] Combination therapy According to the present invention, it is contemplated that a plurality of the probiotic vaccines of the present invention may be administered in combination to a subject in need thereof in a specific therapy for preventing, treating, and / or reducing the risk of cancer, autoimmune diseases, respiratory diseases (e.g., asthma), neurological diseases, infectious diseases, and other diseases (osteoporosis, hypercholesterolemia, etc.), which method comprises administering a plurality of the probiotic vaccines of the present invention described herein. In another embodiment, this specific combination therapy may also be administered in combination with at least one other therapeutic agent.

[0150] The present invention also contemplates the use of a combination of recombinant phages of the present invention in a specific therapy for preventing, treating, and / or reducing the risk of cancer, autoimmune diseases, respiratory diseases (e.g., asthma), neurological diseases, infectious diseases, and other diseases (osteoporosis, hypercholesterolemia, etc.) in a subject in need thereof, the method comprising administering a plurality of recombinant phages of the present invention described herein. In another embodiment, the specific combination therapy can be further combined with at least one other therapeutic agent.

[0151] The present invention also contemplates the use of a probiotic vaccine of the present invention in combination with a recombinant phage of the present invention in a specific therapy for preventing, treating, and / or reducing the risk of cancer, autoimmune diseases, respiratory diseases (e.g., asthma), neurological diseases, infectious diseases, and other diseases (osteoporosis, hypercholesterolemia, etc.) to a subject in need thereof, which method comprises administering a combination of both a probiotic vaccine described herein and a recombinant phage described herein. In another embodiment, this specific combination therapy can also be used in combination with at least one other therapeutic agent.

[0152] The present invention also contemplates that the independent methods of administering the probiotic vaccines of the present invention for a particular treatment or the recombinant phages of the present invention for a particular treatment can be used in combination with other therapeutic agents for preventing, treating, and / or reducing the risk of cancer, autoimmune diseases, respiratory diseases (e.g., asthma), neurological diseases, infectious diseases, and other diseases (osteoporosis, hypercholesterolemia, etc.), including administering to a subject in need thereof the probiotic vaccines of the present invention, recombinant phages, or both, in combination with the other therapeutic agents. Thus, probiotic vaccinations of the present invention and / or direct administration of recombinant bacteriophages of the present invention (for vaccination or treatment) can be used in combination with other therapeutic agents.

[0153] For example, subjects administered at least one probiotic vaccine and / or at least one recombinant bacteriophage of the present invention can also receive chemotherapy, immuno-oncology therapy, radiation therapy, and / or surgery before, simultaneously with, or after direct vaccination and / or probiotic vaccination. Chemotherapy is used to shrink or slow cancer growth. Chemotherapy is recommended for many cancers after initial surgery, but chemotherapy is sometimes administered before surgery to shrink the cancer. The number of cycles of chemotherapy treatment depends on the stage of the disease. Chemotherapy may neutralize the anti-tumor immune response generated by vaccine therapy. Furthermore, chemotherapy can be safely combined with immunotherapy, possibly with additive or synergistic effects, as long as the combination is rationally designed. Examples of chemotherapeutic agents that can be used to treat cancer subjects include, but are not limited to, carboplatin, cisplatin, cyclophosphamide, docetaxel, doxorubicin, etoposide, gemcitabine, oxaliplatin, paclitaxel, Taxol™, topotecan, vinorelbine, etc. In some embodiments, the subject receiving the probiotic vaccination is undergoing chemotherapy, radiation therapy, and / or surgical treatment for gynecological or peritoneal cancer. Immunotherapy (immuno-oncology therapy) refers to the induction, promotion, and / or modulation of an immune response by eliciting the production of antibodies against selected targets, such as TNF. Immunotherapies used in combination with the probiotic vaccines of the present invention (or the recombinant phage of the present invention) include treatment with Keytruda® (pembrolizumab), Opdivo® (nivolumab), etc.

[0154] In addition to or apart from chemotherapy, subjects receiving the probiotic vaccine of the present invention can be treated with any other treatment that is beneficial for specific cancers.For example, subjects with ovarian cancer, fallopian tube cancer, or peritoneal cancer can be treated with a COX-2 inhibitor before, simultaneously with, or after receiving the probiotic vaccine of the present invention, as described in WO2005 / 037995 by Yu and Akasaki.In another embodiment, subjects receiving the probiotic vaccine of the present invention can be treated with bevacizumab (Avastin®) before, simultaneously, or after receiving the probiotic vaccine. [Example]

[0155] The following examples are provided to better illustrate the claimed disclosure and should not be construed as limiting the scope of the disclosure. Where specific materials are mentioned, they are for illustrative purposes only and are not intended to limit the disclosure. Those skilled in the art may develop equivalent means or reactants without the exercise of inventive capacity and without departing from the scope of the disclosure.

[0156] Example 1 Construction of PD-1 / AFP10 epitope-containing Nissle probiotic vaccine (M13KE-SALV-0004): An mRNA sequence encoding the amino acid sequence -AFPEDRSQPG- (SEQ ID NO: 8; referred to herein as "AFP10" and "SALV-0004") was inserted between the pIII signal peptide and pIII protein coding sequence of M13KE RF I DNA to construct M13KE-AFP 10 RF I DNA. Sequence information of the recombination insertion site within the M13KE phage is shown in Figure 5.

[0157] Preparation of M13KE vector Logarithmic-phase Escherichia coli ER2738 strains were infected with M13KE phage. After incubation, the cells were harvested by centrifugation, and the plasmid was extracted using the Axygen MidiPrep Plasmid Extraction Kit. M13KE RF I DNA was further purified and then loaded onto an agarose gel for QC.

[0158] M13KE RF I DNA was digested with Kpn1 / EagI and separated by agarose gel electrophoresis.

[0159] Construction of M13KE-SALV-0004 phage DNA encoding the AFP10 epitope (SALV-0004; SEQ ID NO: 8) was synthesized, digested with KpnI and EagI, and ligated to M13KE RF I DNA. The AFP10-M13KE RF I DNA construct was transformed into competent Escherichia coli ER2738 cells. After resuscitation, logarithmic-phase E. coli ER2738 was infected and cultured overnight at 32°C on TOP-Agar LB (containing IPTG / X-gal) plates. Many blue spots were observed on the plates where the recombinant M13KE-AFP10 phage had been transformed into E. coli ER2738, whereas no blue spots were observed in the control group. Several blue spots were selected and verified by sequencing. The results indicated that all clones contained the correct construct corresponding to the correct mRNA sequence.

[0160] Phage preparation Logarithmic growth phase Escherichia coli ER2738 strain was infected with M13KE phage carrying the correct AFP10 epitope sequence (SALV-0004; SEQ ID NO: 8). After overnight incubation, the culture supernatant was collected, glycerol was added to a final concentration of 10%, and stored at -20°C.

[0161] The recombinant M13KE-SALV-0004(AFP10) phage of the present invention described above was used to infect Escherichia coli Nissle 1917 strain and used as a probiotic, referred to herein as "N4" probiotic vaccine. These Nissle bacteria infected with bacteriophage MK13E-SALV-0004 expressing the AFP10 epitope were then used as a probiotic oral vaccine to induce the production of anti-PD-1 antibodies in subjects against cancers such as gastric cancer, melanoma, non-small cell lung cancer (NSCLC), head and neck squamous cell carcinoma (HNSCC), urothelial carcinoma, non-muscle-invasive bladder cancer (NMIBC), colon or rectal cancer, esophageal or certain gastroesophageal junction (GEJ) cancers, cervical cancer, renal cell carcinoma (RCC), advanced endometrial cancer, cutaneous squamous cell carcinoma (cSCC), and / or triple-negative breast cancer (TNBC). Nissle probiotic phage (i.e., phage + E. coli) infected with M113KE-AFP10, which contains the epitope described above, was then produced. Oral administration of M113KE-AFP10-infected Nissle probiotics was expected to produce both SlgA and IgG anti-PD-1 antibodies in the subjects, whereas intravenous injection of the recombinant phage alone (separate from the probiotic bacteria) resulted in the production of IgA and IgG in the subjects, which showed reactivity to PD-1 on human cancer cells in the subjects.

[0162] Example 2 Construction of a HER2 epitope-containing probiotic vaccine (EcN's M13KE-SALV-0006): A bacteriophage oral vaccine containing a functionally expressed HER2 epitope on the M13KE phage infected cells of the Escherichia coli Nissle 1917 strain was developed. Specifically, the HER2 epitope was incorporated into the MKE, fl, and / or pIII coat protein genes of the M13 bacteriophage to produce a vaccine that overcomes immune tolerance to HER2. The carrier acted as an adjuvant, improving epitope stability and presentation to B cells.

[0163] Antibodies against two distinct regions, the N-terminus and C-terminus of the fragment, were found to exhibit growth inhibition. Epitope mapping of the C-terminal antibody revealed a 24-amino acid region (LPESFDGDPASNTAPLQPEQLQVF; SEQ ID NO: 1) with two distinct epitopes that mediated efficient growth inhibition. These results demonstrate that antibodies targeting this region of HER2 domain III, unlike the well-known monoclonal antibodies trastuzumab and pertuzumab, bind to HER2 on live cells and exert growth inhibition.

[0164] In a specific embodiment of the invention described herein, the P4378-394 B-cell epitope (PESFDGDPASNTAPLQPEQLQ; SEQ ID NO: 2) derived from the HER2 extracellular domain was displayed by insertion into gene III (i.e., pIII) of a filamentous phage. Previous preclinical studies have shown that immunization with the P4378-394 epitope, either as a single epitope or as a multi-epitope formulation, induces HER2-specific IgG antibodies with potent antitumor activity. Furthermore, a phase I clinical trial using a multi-epitope vaccine containing the P4378-394 epitope demonstrated that the vaccine was safe, well-tolerated, and effective in overcoming immune tolerance to HER2.

[0165] M13KE-HER2RFI DNA was constructed by inserting an mRNA sequence encoding the amino acid sequence -ESFDGDPASNTAPLQPEQL- (SEQ ID NO: 38) between the pIII signal peptide and pIII protein coding sequence of M13KE-HER2RFI DNA.

[0166] Preparation of M13KE vector Logarithmic-phase Escherichia coli ER2738 strains were infected with M13KE phage. After incubation, the cells were harvested by centrifugation, and the plasmid was extracted using the Axygen MidiPrep Plasmid Extraction Kit. M13KE RF I DNA was further purified and then loaded onto an agarose gel for QC.

[0167] M13KE RF I DNA was digested with Kpn1 / EagI and separated by agarose gel electrophoresis.

[0168] Construction of recombinant M13KE-SALV-0006 phage DNA encoding the HER2 epitope (SALV-0006; SEQ ID NO: 38) was synthesized, digested with Kpn1 / EagI, and ligated to M13KE RF I DNA. The HER2-M13KE RF I DNA construct was transformed into competent Escherichia coli ER2738 strain cells. After resuscitation, logarithmic-phase Escherichia coli ER2738 strains were infected and cultured overnight at 32°C on TOP-Agar LB (containing IPTG / X-gal) plates. The clones transformed with the M13KE-HER2 phage (SEQ ID NO: 38) of the present invention into Escherichia coli ER2738 strain showed numerous blue spots on the plates, whereas no blue spots were observed in the control group. Several blue spots were selected and verified by sequencing. The results indicated that all clones contained the correct construct corresponding to the mRNA sequence. GAATCATTTGACGGAGATCCCGCTAGTAACACCGCGCCGCTGCAGCCGGAGCAGTTG.

[0169] Phage preparation Logarithmic growth phase Escherichia coli ER2738 strain was infected with M13KE phage carrying the correct HER2 epitope sequence (M13KE-SALV-0006 phage; SEQ ID NO: 38). After overnight incubation, the culture supernatant was collected, supplemented with 10% glycerol, and stored at -20°C.

[0170] The M13KE-HER2 phage described above was used to infect Escherichia coli Nissle 1917 strains, which were identified as male and used as probiotics. Nissle bacteria of the present invention infected with a bacteriophage expressing a HER2 epitope (referred to herein as "N6" and comprising SEQ ID NO: 38) are nonlytic and continuously secrete (or excrete) the SALV-0006 recombinant phage, allowing them to be used as a probiotic oral vaccine to continuously promote the production of anti-HER2 antibodies against cancer (e.g., breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, and / or gastric cancer) in patients / subjects. M13KE-HER2-infected Nissle probiotic phage (e.g., phage + Escherichia coli) containing the epitope described above is generated. In the present invention, oral administration of Nissle probiotic phage infected with M13KE-HER2 is expected to produce secretory antibodies (SlgA) and IgG antibodies in patients, whereas injection of recombinant phage alone (separate from probiotic bacteria) will produce IgG in patients with reactivity against HER2 on the patient's human cancer cells.

[0171] Example 3 Construction of SALV-0013 Probiotic Vaccine: SALV-0013, corresponding to the peptide epitope SEQ ID NO:29, was inserted into the MK13 phage as described above to form the recombinant bacteriophage MK13 / SALV-0013. Bacteriophage MK13 / SALV-0013 was then used to infect cells of Nissle 1917 and ER2738 strains to generate the "N13" and "ER13" recombinant phage-infected E. coli strains, respectively.

[0172] An mRNA sequence encoding the amino acid sequence -ESKVDPSKAW- (SEQ ID NO: 29; referred to herein as "SALV-0013") was inserted between the pill signal peptide and pill protein coding sequence of M13KE RF I DNA to construct M13KE-SALV-0013RF I DNA. Sequence information for the recombination insertion site within the M13KE phage is shown in Figure 6.

[0173] Preparation of M13KE vector Logarithmic-phase Escherichia coli TG1 / ER2738 strains were infected with M13KE phage. After incubation, the cells were harvested by centrifugation, and the plasmid was extracted using the Axygen MidiPrep Plasmid Extraction Kit. M13KE RF I DNA was further purified and then loaded onto an agarose gel for QC.

[0174] M13KE RF I DNA was digested with Kpn1 / EagI and separated by agarose gel electrophoresis.

[0175] Construction of M13KE-SALV-0013 recombinant phage DNA encoding the SALV-0013-epitope (SEQ ID NO: 29) was synthesized, digested with KpnI / EagI, and ligated with M13KE RF I DNA. The SALV-0013-M13KE RF I DNA construct was transformed into competent Escherichia coli TG1 cells. After resuscitation, logarithmic-phase E. coli TG1 was infected and cultured overnight at 32°C on TOP-Agar LB (containing IPTG / X-gal) plates. Many blue spots were observed on plates where the recombinant M13KE-SALV-0013 phage had been transformed into E. coli TG1, whereas no blue spots were observed in the control group. Several blue spots were selected and verified by sequencing. The results indicated that all clones contained the correct construct corresponding to the correct mRNA sequence.

[0176] Phage preparation Logarithmic growth phase Escherichia coli TG1 strain was infected with M13KE-SALV-0013 phage containing the correct SALV-0013 epitope sequence. After overnight incubation, the culture supernatant was collected, glycerol was added to a final concentration of 10%, and the culture was stored at -20°C. The titer was determined by gradient dilution and was 1.36 × 10 10 pfu / mL.

[0177] The recombinant M13KE-SALV-0013 phage of the present invention described above was used to infect Escherichia coli Nissle 1917 strains to produce probiotic vaccines of the present invention that can be used as probiotic therapies. For example, this infected Nissle strain carrying bacteriophage expressing the SALV-0013 epitope was then used as a probiotic oral vaccine to generate anti-PMSA antibodies against cancers such as prostate cancer, melanoma, non-small cell lung cancer (NSCLC), head and neck squamous cell carcinoma (HNSCC), urothelial carcinoma, non-muscle-invasive bladder cancer (NMIBC), colon or rectal cancer, esophageal cancer or certain gastroesophageal junction (GEJ) cancers, cervical cancer, renal cell carcinoma (RCC), advanced endometrial cancer, cutaneous squamous cell carcinoma (cSCC), and / or triple-negative breast cancer (TNBC). We produced Nissle probiotic phage (i.e., phage + E. coli) infected with M113KE-SALV-0013 containing the epitope described above. In animal experiments described in the Examples, oral administration of Nissle probiotics infected with M113KE-SALV-00013 was confirmed to produce both SlgA and IgG anti-PSMA antibodies. Meanwhile, intravenous injection or infusion of the recombinant phage alone (separate from the probiotic bacteria) resulted in the production of both IgA and IgG antibodies in the subject's body, demonstrating reactivity to PSMA on the subject's human cancer cells.

[0178] Example 4 Construction of a Nissle probiotic vaccine containing CD38 epitope: M13KE-CD38 RF I DNA was constructed by inserting an mRNA sequence encoding one of the amino acid sequences QPEKVQTLEAWVIHGG (SEQ ID NO: 3), ISKRNIQFSCKNIYR (SEQ ID NO: 4), TFGSVEVHNL (SEQ ID NO: 5), QTLEA (SEQ ID NO: 6), IQFSC (SEQ ID NO: 7), KRNIQFSCKNI (SEQ ID NO: 49), QTLEAQTLEA (SEQ ID NO: 58), and IQFSCIQFSC (SEQ ID NO: 59) between the pIII signal peptide and pIII protein coding sequence of M13KE RF I DNA.

[0179] Preparation of M13KE vector Logarithmic-phase Escherichia coli ER2738 strains were infected with M13KE phage. After incubation, the cells were harvested by centrifugation and the plasmid was extracted using the Axygen MidiPrep Plasmid Extraction Kit. M13KE RF I DNA was further purified and loaded onto an agarose gel for QC.

[0180] M13KE RF I DNA was digested with Kpn1 / EagI and separated by agarose gel electrophoresis.

[0181] Construction of M13KE-CD38 phage DNA encoding the CD38 epitope (encoding QPEKVQTLEAWVIHGG (SEQ ID NO: 3), ISKRNIQFSCKNIYR (SEQ ID NO: 4), TFGSVEVHNL (SEQ ID NO: 5), QTLEA (SEQ ID NO: 6), IQFSC (SEQ ID NO: 7), KRNIQFSCKNI (SEQ ID NO: 49), QTLEAQTLEA (SEQ ID NO: 58), and IQFSCIQFSC (SEQ ID NO: 59)) was synthesized, digested with KpnI / EagI, and ligated to M13KE RF I DNA. The HER2-M13KE RF I DNA construct was transformed into Escherichia coli ER2738 competent cells. After resuscitation, logarithmic-phase E. coli ER2738 strains were infected and cultured overnight at 32°C on TOP-Agar LB (containing IPTG / X-gal) plates. Many blue spots were observed on plates where M13KE-CD38 was transformed into Escherichia coli ER2738, whereas no blue spots were observed in the control group. Several blue spots were selected and verified by sequencing. The results showed that all clones contained the correct constructs corresponding to the mRNA sequences encoding QPEKVQTLEAWVIHGG (SEQ ID NO: 3), ISKRNIQFSCKNIYR (SEQ ID NO: 4), TFGSVEVHNL (SEQ ID NO: 5), QTLEA (SEQ ID NO: 6), IQFSC (SEQ ID NO: 7), KRNIQFSCKNI (SEQ ID NO: 49), QTLEAQTLEA (SEQ ID NO: 58), and IQFSCIQFSC (SEQ ID NO: 59).

[0182] Phage preparation Logarithmic-phase Escherichia coli ER2738 strain was infected with M13KE phage carrying the correct CD38 exogenous peptide epitope. After overnight incubation, the culture supernatant was collected, glycerol was added to a final concentration of 10%, and stored at -20°C.

[0183] The M13KE-CD38 phage described above was used to infect Escherichia coli Nissle 1917 strains as a probiotic. These Nissle-infected bacteria, containing bacteriophages expressing the CD38 epitope, were then used as a probiotic oral vaccine to generate anti-CD38 antibodies against cancers (e.g., multiple myeloma) in subjects. Nissle probiotic phage (i.e., phage + E. coli) infected with M113KE-CD38 containing the epitope described above was produced. Oral administration of Nissle probiotics infected with M113KE-CD38 is expected to generate both SlgA and IgG anti-CD38 antibodies in patients. Meanwhile, injection of the recombinant phage alone (separate from the probiotic bacteria) resulted in the production of IgG in the subject's body, which showed reactivity to CD38 on the patient's human cancer cells.

[0184] Example 5 Preparation of Oral Probiotic LB Formulation: Two milliliters of concentrated Nissle strain from the starter batch of Escherichia coli Nissle 1917 (EcN) strain was added to a 50-ml flask of LB medium and cultured in a shaking incubator at 37°C and 140 rpm for 4 hours. Next, 20 μl of the recombinant phage particles of the present invention (e.g., SALV-0004, SALV-0006, and SALV-0013) was added to the Nissle bacteria and cultured at 37°C and 140 rpm for 4 hours. The infected host bacterial culture was then added to 400 ml of LB medium and cultured at 37°C and 140 rpm for 4 hours. The culture was then dispensed into 20-ml aliquots in 50-ml test tubes and centrifuged at 3,000 rpm for 10 minutes. Two milliliters of the supernatant was collected using a pipette, and the remaining supernatant was returned to the flask, leaving the pellet in the test tube. Next, 2 ml of the supernatant was returned to the test tube and vortexed at 600 rpm until the pellet dissolved, producing a bacteriophage concentrate. Using a pipette, 2 ml of concentrated bacteriophage was collected from the 50 ml tube and transferred to a 2 ml test tube for storage. The resulting concentrate was frozen and stored at -20°C until use.

[0185] Example 6 Preparation of IV Therapy Formulation Starting with 2 ml of concentrated probiotic recombinant bacteriophage obtained as described herein, 2 ml of the concentrate was centrifuged at 10,000 rpm for 15 minutes. The supernatant was drawn into a 3 ml syringe, and the pellet was left in a test tube. The needle was removed from the syringe, a 0.2 μm Swinnex filter was attached, and the supernatant was drawn through the filter into a 2 ml test tube. The resulting filtered supernatant was ready for either intravenous or nasal administration.

[0186] Example 7 Preparation of an Oral Probiotic Coconut Water Formulation: Two milliliters of concentrated Nissle strain from a starter batch of Escherichia coli Nissle 1917 (EcN) strain was added to coconut water in a 50-ml flask and cultured in a shaking incubator at 37°C and 140 rpm for 4 hours. Next, 20 μl of recombinant phage particles of the present invention (e.g., SALV-0004, SALV-0006, and SALV-0013) was added to the Nissle bacteria / coconut water solution in the 50-ml flask and cultured at 37°C and 140 rpm for 4 hours. The infected Nissle bacteria solution from the 50-ml flask was then added to 400 ml of coconut water in a larger flask and cultured at 37°C and 140 rpm for 4 hours. Next, 20 ml of the 400 ml solution was aliquoted into a 50-ml test tube and centrifuged at 3,000 rpm for 10 minutes. Two milliliters of the supernatant was removed using a pipette, and the remaining supernatant was returned to the 400-ml flask, leaving the pellet in the test tube. Two milliliters of the supernatant was then returned to the test tube and vortexed at 600 rpm until the pellet dissolved, producing a bacteriophage concentrate in coconut water. Using a pipette, 2 milliliters of concentrated bacteriophage was removed from the 50-ml tube and transferred to a 2-ml tube for storage. The concentrate dissolved in coconut water was frozen at -20°C until use.

[0187] SALV-006 ("N6") infected with Nissle 1917 strain was found to grow at approximately equivalent rates in both coconut water and LB broth. Plating experiments comparing 10-fold dilutions of coconut water and LB broth showed that coconut water medium produced significantly higher plaque-forming units than LB broth, demonstrating equivalence. The inventive method of culturing the inventive probiotic bacterial therapeutic in coconut water advantageously utilizes coconut water as a delivery medium, offering significant cost and logistical advantages over prior art methods.

[0188] Example 8 Preparation of a probiotic vaccine gummy formulation for oral administration To prepare Luria-Bertani (LB) broth, 400 ml of distilled water was mixed with 10 g of HIMEDIA Luria-Bertani broth (Miller) and heated to 37°C on a heating plate equipped with a magnetic mixer. From this LB broth, 20 aliquots of treatment drug were prepared, sufficient for three doses in a test group of four animals (e.g., rats): two for titration and six for pre-dose.

[0189] To inoculate LB medium, 2 ml vials of starter batches of each of the inventive probiotic Nissle bacterial vaccines (e.g., SALV-0004 (N4)), SALV-0006 (N6), and SALV-00013 (N13)) were thawed at room temperature and mixed with LB medium. After thawing, the starter batches were gently vortexed and added to the LB medium. To cultivate the bacteria of the present invention, including the recombinant bacteriophages of the present invention (SALV-0004, SALV-0006, and SALV-00013), the culture medium infected with the starter batches was placed in a shaking incubator and cultured at 37°C and 140 rpm for 4 to 8 hours. The medium began to become cloudy after a few hours and became nearly opaque after 5 hours.

[0190] To prepare the gummy formulation, 250 ml of Welch's grape juice, 1 tablespoon of sugar, and 2 teaspoons of agar powder were mixed and stirred on a hot plate for 2 minutes until boiling. Once boiling, the mixture was removed from the heat and allowed to cool. 1.5 ml of the mixture was pipetted into 5 ml test tubes in a preheated warming block.

[0191] To prepare treatment aliquots for animal testing, 20 ml aliquots were taken from each tube and centrifuged at 3,500 rpm for 10 minutes. 500 μl was removed from each tube, the supernatant was discarded, and 500 μl was returned to the respective tube. Each tube was vortexed until the pellet was broken up. 500 μl of the liquefied pellet was pipetted and added to 1.5 ml of the gummy formulation in a warming tray.

[0192] To plate and mold the therapeutic gummy formulations, 500 μL of liquefied pellets were added to 1.5 mL of gummy formulation, and each 5 mL tube was capped and shaken. The mixture was then briefly vortexed to thoroughly mix and tapped to remove air bubbles. The mixture was poured into gummy molds and refrigerated until ready to dispense.

[0193] Example 9 Generation of target IgA and IgG responses: In this example, oral formulations of ER6, N6, and N13 probiotic bacterial vaccines, gummy formulations of N6G, and recombinant phage of the present invention obtained from the supernatant of the N6 batch were filtered as described herein and intravenously administered as described herein. These formulations were administered to rats orally, as oral gummies, or intravenously.

[0194] The left graph in Figure 2A shows that intravenous administration of "IV6" therapy, corresponding to the recombinant phage MK13E-SALV-0006 sequence of the present invention in an intravenous formulation, demonstrated a statistically significant response to IgA throughout the entire period following injection.

[0195] The graph on the right in Figure 2B shows that by day 17, the blank phage (unmodified) and control groups were the two lowest groups in terms of serum IgG antibody production. It also shows that IV6, the only formulation injected IV, produced the highest statistically significant response for IgG on days 10 and 17.

[0196] Example 10 Orally Administered SALV-0013 Elicits a Significant SIgA Response Figure 3 shows that in animal experiments, N13 (Escherichia coli Nissle 1917 strain infected with recombinant phage M13KE-SALV-0013) orally administered via the gummy formulation of the present invention elicited an overwhelming SIgA immune response. The delivery mechanism of the present invention, represented by the "N" series of data points, demonstrated a significant response compared to the control upon oral administration. Secretory IgA (SIgA) plays an important role in protecting and regulating homeostasis of the intestinal, respiratory, and urogenital mucosal epithelia, which separate the internal environment from the external environment. Orally delivered Nissle 1917 strain-hosted treatments of the present invention (e.g., N13, N6, and N4) demonstrated significantly higher levels of SIgA than the control group. Furthermore, the control blank phage did not elicit an SIgA response, indicating that it is inactive without modification. It is also noteworthy that N13 (e.g., SALV-0013; ESKVDPSKAW), which is only 10 aa long, actually showed the strongest SIgA response, indicating that shorter 10 aa sequences can generate significant secretory immune responses.

[0197] Furthermore, these experimental results demonstrate that the gummy formulation of N6G is more effective than oral administration and is a practical administration form. These results also demonstrate the ability of the probiotic vaccine of the present invention (e.g., SALV-0013) to penetrate mucosal barriers and elicit an immune response when administered orally.

[0198] As shown in Example 11 below, when administered in combination with both oral and IV formulations, in a preclinical prostate cancer xenograft mouse study, animals treated with SALV-0013 were observed to have significantly reduced tumor size compared to the control group and a similar oral / IV combination of SALV-0004 and SALV-0006.

[0199] As a result, the probiotic vaccine of the present invention (SALV-0013) can be administered orally, penetrate the mucosa, deliver trillions of target epitopes, and induce an immune response.

[0200] Example 11 Prostate Cancer Xenograft Athymic Mouse Study: Athymic nude mice were implanted with the LNCaP prostate cancer cell line. The LNCaP cell line is an epithelial cell line derived from human prostate cancer. The FGC (Fast Growth Colony) cell line, a derivative of the LNCaP cell line, possesses all of the major characteristics reported for the original LNCaP cultured cells, including androgen dependence, and is considered a standard in the art. See, e.g., Steenbrugge et al., Urol Res. 1989;17(2):71-7. doi: 10.1007 / BF00262024.

[0201] In this example, three recombinant phages of the present invention were tested in a combined IV / oral therapy against a human prostate cancer cell line injected into immunodeficient mice. SALV-0004 (SEQ ID NO: 8), SALV-0006 (SEQ ID NO: 38), and SALV-0013 (SEQ ID NO: 29) were each administered at approximately 1.5 x 10 10 pfu / ml were administered orally and intravenously as follows:

[0202] Oral administration was performed at 0.25 ml per day for 3 days. Intravenous administration was performed as a single dose of 0.15 ml injected over approximately 1 minute on Day 1. For intravenous therapeutic recombinant bacteriophage, the therapeutic agent was prepared by filtering the orally administered bacteriophage through a 0.2 μm filter.

[0203] These mice were athymic, meaning that their thymus was underdeveloped and they were unable to produce T cells that are beneficial for tumor treatment. The results are shown in Figure 4, which demonstrate that SALV-0004, SALV-0006, and SALV-0013 each produced a favorable anti-cancer response. In particular, the SALV-0013 (SEQ ID NO: 29) group showed a 42% reduction in mean tumor size compared to the control group, and no mortality, behavioral abnormalities, or weight loss were observed.

[0204] Example 12 Construction of TNF / VRS10 and TNF / RSS10 epitope-containing Nissle probiotic vaccines (f88-SALV-1001 and f88-SALV-1002): VRS10 and RSS10 DNA were chemically synthesized with a codon-optimized design for Escherichia coli. The mRNA sequences encoding the amino acid sequence -VRSSSRTPSD- (SEQ ID NO: 41; herein referred to as "VRS10" and "SALV-1001") and the amino acid sequence -RSSSRTPSDK- (SEQ ID NO: 42; herein referred to as "RSS10" and "SALV-1002") were inserted between the signal peptide at the 5' end of f88-4pVIII and the pVIII coding sequence using restriction enzymes HindIII / PstI to construct f88-4-VRS10 and F88-4-RSS10 DNA. The sequence information for the recombination insertion site within the f88-4 phage is as follows: f88-4-VRS10: (signal peptide-VRS10-pVIII) MKKSLLVLKASVAVATLVPMLSFA-VRSSSRTPSD-PAEGDDPAKAAFDSLQASATEYIGYAWAMVVVIVGATIGIKLFKKFTSKAS f88-4-RSS10: (signal peptide-VRS10-pVIII) MKKSLLVLKASVAVATLVPMLSFA-RSSSRTPSDK-PAEGDDPAKAAFDSLQASATEYIGYAWAMVVVIVGATIGIKLFKKFTSKAS

[0205] Preparation of f88-4 vector Logarithmic-phase Escherichia coli ER2738 strains were infected with f88-4 phage. After incubation, the cells were harvested by centrifugation and the plasmid was extracted using the Axygen MidiPrep Plasmid Extraction Kit. F88-4 RF I DNA was further purified and loaded onto an agarose gel for QC.

[0206] F88-4 RF I DNA was digested with HindIII / PstI and separated by agarose gel electrophoresis.

[0207] Construction of f88-4-SALV-1001 and f88-4-SALV-1002 phages DNA encoding the VRS10 epitope (SALV-1001; SEQ ID NO: 41) and RSS10 epitope (SALV-1002; SEQ ID NO: 42) was synthesized, digested with HindIII / PstI, and ligated to f88-4 RF I DNA. The VRS10-f88-4 RF I DNA and RSS10-f88-4 RF I DNA constructs were independently transformed into Escherichia coli ER2738 competent cells. After resuscitation, logarithmic-phase E. coli ER2738 strains were infected and grown overnight at 32°C on TOP-Agar LB (containing IPTG / X-gal) plates. The results showed that many blue spots were observed on plates where the recombinant f88-4-VRS10 and f88-4-RSS10 phages were transformed into Escherichia coli ER2738, whereas no blue spots were observed in the control group. Several blue spots were selected and verified by sequencing. The results showed that all clones contained the correct construct corresponding to the correct mRNA sequence.

[0208] Phage preparation Logarithmic growth phase Escherichia coli ER2738 strain was infected with f88-4 phage carrying the correct VRS10 epitope sequence (SALV-1001; SEQ ID NO: 41) or RSS10 epitope sequence (SALV-1002; SEQ ID NO: 42). After overnight incubation, the culture supernatant was collected, glycerol was added to a final concentration of 10%, and the culture was stored at -20°C. Titers were measured by gradient dilution, yielding 8.6 × 10 for f88-4-VRS10. 11 pfu / mL, and 2.9 × 10 for f88-4-RSS10. 12 pfu / mL.

[0209] The recombinant f88-4-SALV-1001 (VRS10) and f88-4-SALV-1002 (RSS10) phages of the present invention described above were used to infect Escherichia coli Nissle 1917 strain to produce "N-f88-1001" (E. coli Nissle 1917 strain infected with recombinant phage f88-4-SALV-1001) and "N-f88-1002" (E. coli Nissle 1917 strain infected with recombinant phage f88-4-SALV-1002), respectively, which were used as probiotic vaccines in the methods of the present invention described herein.

[0210] The foregoing description of the disclosed embodiments is provided to enable those skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles described herein may be applied to alternative embodiments without departing from the spirit or scope of the present invention. It should therefore be understood that the description and drawings set forth herein represent presently preferred embodiments of the invention and, therefore, are representative of the subject matter broadly contemplated by the present invention. It should also be understood that the scope of the present invention fully encompasses alternative embodiments that may become apparent to those skilled in the art, and therefore, is not limited in scope.

Claims

1. 1. A method for preventing, treating and / or reducing the risk of cancer, an autoimmune disease, a respiratory disease, a neurological disease and / or an infectious disease, comprising administering to a subject in need thereof: a. an injectable composition comprising a recombinant phage or probiotic vaccine comprising at least one exogenous peptide epitope corresponding to SEQ ID NOs: 9-133, or a fragment or variant thereof; and b. Oral compositions comprising recombinant phage or probiotic vaccines comprising at least one exogenous peptide epitope corresponding to SEQ ID NOs: 9-133, or a fragment or variant thereof. The method comprises administering a combination of:

2. 10. A method for preventing, treating and / or reducing the risk of cancer, autoimmune diseases, respiratory diseases, neurological diseases and / or infectious diseases, the method comprising administering to a subject in need thereof the probiotic vaccine of claim 5.

3. 3. The method of claim 2, wherein the probiotic vaccine is administered orally.

4. The method of claims 2-3, further comprising administering a recombinant bacteriophage selected from claim 15.

5. The method of claim 4, wherein the recombinant phage is administered intravenously or intramuscularly.

6. 2. The method of claim 1, wherein secretory IgA (SIgA) is produced.

7. 3. The method of claim 1 or 2, wherein secretory IgA (SIgA) is produced in an amount that is 2-100 fold, 2-90 fold, 2-80 fold, 2-70 fold, 2-60 fold, 2-50 fold, 2-40 fold, 2-30 fold, 2-20 fold, 2-10 fold, 2-9 fold, 2-8 fold, 2-7 fold, 2-6 fold, 2-5 fold, or 2-4 fold greater than the control when compared to a control.

8. 1. A probiotic vaccine comprising a recombinant filamentous phage, The phage: ALLEIASCL (SEQ ID NO: 9), MYPPPY (SEQ ID NO: 10), NPLVA (SEQ ID NO: 11), ESKVDPSK (SEQ ID NO: 12), RSSSRTPS (SEQ ID NO: 13), AGICAS (SEQ ID NO: 14), AG-I-CAS (SEQ ID NO: 15; Ile = "AUA" codon), AKPTAKREAIVKAYG (SEQ ID NO: 16), ISPDGCYSL (SEQ ID NO: 17), LTFDYVVTF (SEQ ID NO: 18), VLYQDVNCT (SEQ ID NO: 19), QQQQGQTITKKSAAEASKKPRQKRTAVLVQDVGCTFSTVFPVTSFGPLVR (SEQ ID NO: 20), VLYQGVNCT (SEQ ID NO: 21), DAPF (SEQ ID NO: 22), KQIVERILEEE (SEQ ID NO: 23), RYLTDMTLEE (SEQ ID NO: 24), DDVKRENPEHEEYNN (SEQ ID NO: 25), RESGSS (SEQ ID NO: 26), FRHDSGY (SEQ ID NO: 27), AGICASAGICAS (SEQ ID NO: 28), ESKVDPSKAW (SEQ ID NO: 29), DAPFKQIVERILEEE (SEQ ID NO: 30), RESGSSRYLTDMTLEE (SEQ ID NO: 31), DDVKRENPEHEEYNN (SEQ ID NO: 32), CVADYSVLY (SEQ ID NO: 33), HVGEIPVAY (SEQ ID NO: 34), NQKLIANQF (NQK-Q8) (SEQ ID NO: 35), NQKLIANAF (NQK-A8) (SEQ ID NO: 36), RVAGDSGFAAY (SEQ ID NO: 37), ESFDGDPASNTAPLQPEQL (SEQ ID NO: 38), EFINVPEGGS (SEQ ID NO: 39), EFINVPEEFINVPE (SEQ ID NO: 40), VRSSSRTPSD (SEQ ID NO: 41), RSSSRTPSDK (SEQ ID NO: 42), MYPPPYMYPPPY (SEQ ID NO: 43), VELMYPPPYYLGIG (SEQ ID NO: 44), LMYPPPYYLGIGN (SEQ ID NO: 45), NPLVANPLVA (SEQ ID NO: 46), LPNAIGRGKWWRP (SEQ ID NO: 47), ETPEGAEA (SEQ ID NO: 48), KRNIQFSCKNI (SEQ ID NO: 49), LTFDY (SEQ ID NO: 50), LTFDYLTFDY (SEQ ID NO: 51), NTVSCNRPHCITE (SEQ ID NO: 52), EERRR (SEQ ID NO: 53), EERRRREERRR (SEQ ID NO: 54), LTHVKIND (SEQ ID NO: 55), NANPNANPNANPNAN (SEQ ID NO: 56), EIRQAGRPNKPDSI (SEQ ID NO: 57), QTLEAQTLEA (SEQ ID NO: 58), IQFSCIQFSC (SEQ ID NO: 59), DAPPFDAPF (SEQ ID NO: 60), RESGSSRESGSS (SEQ ID NO: 61), VVVGADVGKS (SEQ ID NO: 62), PTPEEDQILKK (SEQ ID NO: 63), APKAQIKESLR (SEQ ID NO: 64), ASPLLGSQHFDVPPRQL (SEQ ID NO: 65), QYDSWPHYPE (SEQ ID NO: 66), KVIQAQTAFSANPANPAILS (SEQ ID NO: 67), PIPHDGNLYPRLYPE (SEQ ID NO: 68), SKCRRCRLLD (SEQ ID NO: 69), QNTKCRCKPNFFC (SEQ ID NO: 70), LCTLAWAHPLVGHM (SEQ ID NO: 71), ESVDPKNYPKK (SEQ ID NO: 72), YLSETVSNALGPQGRRLEE (SEQ ID NO: 73), INDPRLRNPSDKFIY (SEQ ID NO: 74), SPRMSGLLSQTGYLSDNHILI (SEQ ID NO: 75), GLPSLGLTL (SEQ ID NO: 76), HRGEFS (SEQ ID NO: 77), HRGEFSHRGEFS (SEQ ID NO: 78), THLHWHTV (SEQ ID NO: 79), VQDYVQMKASELEQ (SEQ ID NO: 80), REIEGRVMVTDPENVPEEDG (SEQ ID NO: 81), AGAFAPSAAVA (SEQ ID NO: 82), GYYYRDDSLPVLLE (SEQ ID NO: 83), EDPAFYMLKGKLQYELQ (SEQ ID NO: 84), ALFDAQAQV (SEQ ID NO: 85), LLYDLTEKQVY (SEQ ID NO: 86), ALDENTKLWYA (SEQ ID NO: 87), VLVESEHQV (SEQ ID NO: 88), APNRTITVDDKMSLRLH (SEQ ID NO: 89), SSDWVTLGTSASLLR (SEQ ID NO: 90), STPYVGLSLSCAANQ (SEQ ID NO: 91), PSPQYSWRINGIPQQHTQVLF (SEQ ID NO: 92), STLYREELMTTLLYYEIFL (SEQ ID NO: 93), CEKNSVLEVIAYSSSSE (SEQ ID NO: 94), NPTTYQMDVNPEGKYS (SEQ ID NO: 95), HTPPLDPQELDILKT (SEQ ID NO: 96), RPKSNIVLLS (SEQ ID NO: 97), SGPKPLFRRMSSLVGPTQ (SEQ ID NO: 98), RPKSNIVLLSAEEKKEQT (SEQ ID NO: 99), DYEDRYYRE (SEQ ID NO: 100), RPLKGAPLPAL (SEQ ID NO: 101), MTEQQWNFAGIEAAASAIQG (SEQ ID NO: 102), KTIAYDEEARR (SEQ ID NO: 103), EEFVVEFDLPGIKA (SEQ ID NO: 104), CVTIMTVDNPASTTNKDKLFAVWKITYKDT (SEQ ID NO: 105), ILKEPVHGV (SEQ ID NO: 106), SLYNTVATL (SEQ ID NO: 107), LSEIKGVIVHRLEGV (SEQ ID NO: 108), MLMAASRAL (SEQ ID NO: 109), RAMRMVYYL (SEQ ID NO: 110), FFVRSIGSKGGKLAAGKYTDAVTV (SEQ ID NO: 111), EEPEESDEDDFGMGGLF (SEQ ID NO: 112), CINGVCWTV (SEQ ID NO: 113), KLVALGINAV (SEQ ID NO: 114), NLVPMVATV (SEQ ID NO: 115), TPRVTGGGAM (SEQ ID NO: 116), TETEPPVAL (SEQ ID NO: 117), REALHEAL (SEQ ID NO: 118), DVIGRIENGERL (SEQ ID NO: 119), RLIKEDVYL (SEQ ID NO: 120), KTLETNPYVK (SEQ ID NO: 121), GRADALRVLSK (SEQ ID NO: 122), LSRELSSLASQPLSR (SEQ ID NO: 123), SLLPDSPVGQ (SEQ ID NO: 124), SPSQPWQRL (SEQ ID NO: 125), FLAEDALNTV (SEQ ID NO: 126), CTHYMQLAC (SEQ ID NO: 127), QEANASAQTKCDIE (SEQ ID NO: 128), VVVGACVGKS (SEQ ID NO: 129), LPAIVREAI (SEQ ID NO: 130), SPRRKLISVDSRSVSL (SEQ ID NO: 131), DKPVYTPDQSVKVRV (SEQ ID NO: 132), and / or DNVLDHLTGRSC (SEQ ID NO: 133); and Bacteria infected with the recombinant phage The probiotic vaccine comprises a nucleic acid encoding a recombinant polypeptide comprising an exogenous peptide epitope selected from the group consisting of:

9. 9. The probiotic vaccine of claim 8, wherein the exogenous peptide epitope is functionally expressed on a coat protein of the phage selected from the group consisting of pIII, pVI, pVII, pVIII and pIX.

10. The probiotic vaccine according to claims 8 to 9, wherein the coat protein is pIII or pVIII.

11. The phage may be selected from the group consisting of filamentous phage, M13, fd, IKe, CTX-φ, Pf1, Pf2, Pf3, f1, M13, MKE, M13KE, type 8, type f88, f88-4, Myoviridae (P1-like viruses, P2-like viruses, Mu-like viruses, SPO1-like viruses, phiH-like viruses), Siphoviridae (λ-like viruses, γ-like viruses, T1-like viruses, T5-like viruses, c2-like viruses, L5-like viruses, psiM1-like viruses, phiC31-like viruses, N15-like viruses), Podoviridae (phi29-like viruses, P22-like viruses, N4-like viruses), Tectiviridae (tectiviruses), Corticoviridae (corticoviruses), Liposhrixviridae (lipoviruses), and the like.

11. The probiotic vaccine according to any one of claims 8 to 10, wherein the probiotic vaccine is selected from the group consisting of Iridaceae (Alphaliphothrixvirus, Betaliphothrixvirus, Gammaliphothrixvirus, Deltaliphothrixvirus), Plasmaviridae (Plasmaviruses), Rudiviridae (Rudiviruses), Fuselloviridae (Fuselloviruses), Inoviridae (Inoviruses, Plectroviruses, M13-like viruses, fd-like viruses), Microviridae (Microviruses, Spiromicroviruses, Bdelomicroviruses, Chlamydiamicroviruses), Leviviridae (Leviviruses, Alloleviviruses) and Cystoviridae (Cystoviruses).

12. The probiotic vaccine according to claims 8 to 11, wherein the phage is M13KE or type 8 or type f88.

13. The bacterium is selected from the group consisting of Escherichia coli (E. coli) Nissle 1917, Escherichia coli (E. coli) ER2738, Bacillus amyloliquefaciens, Bacillus polyfermenticus Bispan strain, Bifidobacterium animalis subsp. lactis BB-12 strain, Bifidobacterium animalis subsp. lactis GPS1209 strain, and Bifidobacterium animalis subsp. lactis. HN019 (DR1064) strain, Bifidobacterium bifidum BB-12 strain, Bifidobacterium bifidum Rosell-71 strain, Bifidobacterium breve M-16V strain, Bifidobacterium longum, Bifidobacterium thermophilum, Lactobacillus acidophilus La-1 strain, Lactobacillus brevis HA-112 strain, Lactobacillus fermentum HA-179 strain, Lactobacillus helveticus Lafti L10 strain, Lactobacillus helveticus Rosell-52 strain, Lactobacillus paracasei Lafti L26 strain, Lactobacillus paracasei subsp. paracasei431 strain, Lactobacillus rhamnosus strain HN001 (DR20), Streptococcus salivarius strain DSM 13084, Streptococcus thermophilus, Bacillus coagulans strain GBI-30,6086, Bifidobacterium animalis subsp. lactis strain BB-12, Bifidobacterium longum subsp. infantis, Escherichia coli Nissle 1917 strain, Lactobacillus acidophilus NCFM strain, Lactobacillus paracasei Stl 1 strain (or NCC2461 strain), Lactobacillus johnsonii Lai strain (also called Lactobacillus LCI strain, Lactobacillus johnsonii NCC533 strain), Lactobacillus plantarum 299v strain, Lactobacillus reuteri ATCC 55730 strain (Lactobacillus reuteri SD2112 strain), Lactobacillus reuteri Protectis strain (DSM 17938 strain, ATCC Lactobacillus reuteri Prodentis (a daughter strain of ATCC strain 55730), Lactobacillus reuteri Prodentis (a combination of DSM 17938 / ATCC strain 55730 and ATCC strain PTA 5289), Lactobacillus rhamnosus GG, Saccharomyces boulardii, Lactobacillusa mixture of Lactobacillus rhamnosus strain GR-1 and Lactobacillus reuteri strain RC-14, a mixture of Lactobacillus acidophilus strain NCFM and Bifidobacterium bifidum strain BB-12, a mixture of Lactobacillus acidophilus strain CL1285 and Lactobacillus casei strain LBC80R, a mixture of Lactobacillus plantarum strain HEAL9 and Lactobacillus paracasei 8700:2 strains, a mixture of Lactobacillus bulgaric ...

13. The probiotic vaccine according to any one of claims 8 to 12, wherein the probiotic strain is selected from the group consisting of a mixture of Lactococcus bulgaricus, Lactococcus thermophiles and Lactobacillus bifidus.

14. The probiotic vaccine according to any one of claims 8 to 13, wherein the bacterium is F factor positive and is selected from Escherichia coli Nissle 1917 strain or Escherichia coli ER2738 strain.

15. The probiotic vaccine of any one of claims 8 to 14, wherein the probiotic vaccine generates both IgG and secretory IgA (SIgA) antibodies that bind to the exogenous peptide epitope.

16. A recombinant filamentous phage comprising a recombinant phage genome comprising a nucleic acid encoding a polypeptide comprising an exogenous peptide epitope selected from the group consisting of SEQ ID NOs: 9-133, or a fragment or variant thereof.

17. 17. The recombinant phage of claim 16, wherein the exogenous peptide epitope is functionally expressed on a coat protein of the phage selected from the group consisting of pIII, pVI, pVII, pVIII and pIX.

18. The recombinant phage according to any one of claims 16 to 17, wherein the coat protein is pill or pVIII.

19. The phage may be selected from the group consisting of M13, fd, IKe, CTX-φ, Pf1, Pf2, Pf3, f1, MKE, M13KE, type 8, type f88, f88-4, Myoviridae (P1-like viruses, P2-like viruses, Mu-like viruses, SPO1-like viruses, phiH-like viruses), Siphoviridae (λ-like viruses, γ-like viruses, T1-like viruses, T5-like viruses, c2-like viruses, L5-like viruses, psiM1-like viruses, phiC31-like viruses, N15-like viruses), Podoviridae (phi29-like viruses, P22-like viruses, N4-like viruses), Tectiviridae (tectiviruses), Corticoviridae (corticoviruses), Liposthrixviridae (alf-like viruses), 19. The recombinant phage of any one of claims 16 to 18, wherein the recombinant phage is selected from the group consisting of filamentous phages including: Lipothrixvirus, Betalipothrixvirus, Gammalipothrixvirus, Deltalipothrixvirus), Plasmaviridae (Plasmaviruses), Rudiviridae (Rudivruridae), Fuselloviridae (Fuselloviruses), Inoviridae (Inoviruses, Plectroviruses, M13-like viruses, fd-like viruses), Microviridae (Microviruses, Spiromicroviruses, Bdelomicroviruses, Chlamydiamicroviruses), Leviviridae (Leviviruses, Alloleviviruses) and Cystoviridae (Cystoviruses).

20. The recombinant phage according to claims 16 to 19, wherein the phage is a filamentous phage selected from the group consisting of M13, fd, IKe, CTX-φ, Pf1, Pf2, Pf3, fl, MKE, M13KE, type 8, type f88, and f88-4.

21. The recombinant phage according to any one of claims 16 to 20, wherein the phage is M13KE or f88-4.

22. The recombinant phage of any one of claims 16 to 21, wherein the recombinant phage produces an IgG antibody that binds to an exogenous peptide epitope.

23. 16. A method for preventing or treating cancer, comprising administering to a subject in need thereof one or more of the probiotic vaccines of any one of claims 8-15, or one or more of the recombinant phages of any one of claims 9-15, wherein the exogenous peptide epitope is selected from the group consisting of SEQ ID NOs: 9-12, 29, 38, 43-46, 49, 58, 59, 62-70, 73, 75, 84, 90-92, 95, 96, 101, 117-123, 126, 129 and / or 131.

24. The cancer may be multiple myeloma, epithelial cancer, epithelial ovarian cancer, mucosal melanoma, non-small cell lung cancer, melanoma, head and neck cancer, renal cell carcinoma, Hodgkin's lymphoma, cutaneous squamous cell carcinoma, glioblastoma, esophageal cancer, gastric cancer, duodenal cancer, small intestine cancer, appendix cancer, large intestine cancer, colon cancer, rectal cancer, colorectal cancer, anal cancer, pancreatic cancer, liver cancer, gallbladder cancer, spleen cancer, kidney cancer, bladder cancer, prostate cancer, testicular cancer, uterine cancer, endometrial cancer, ovarian cancer, vaginal cancer, vulvar cancer, breast cancer, lung cancer, thyroid cancer, thymus cancer, brain cancer, nervous system cancer, glioma, oral cancer, 24. The method of claim 23, wherein the cancer is selected from the group consisting of cavity cancer, skin cancer, blood cancer, lymphoma, eye cancer, bone cancer, bone marrow cancer, muscle cancer, non-small cell lung cancer (NSCLC), head and neck squamous cell carcinoma (HNSCC), urothelial cancer, non-muscle invasive bladder cancer (NMIBC), colon or rectal cancer, esophageal cancer or certain gastroesophageal junction (GEJ) cancers, cervical cancer, renal cell carcinoma (RCC), advanced endometrial cancer, cutaneous squamous cell carcinoma (cSCC) and / or triple-negative breast cancer (TNBC).

25. 25. The method of claim 23 or 24, wherein the cancer is multiple myeloma.

26. A method for preventing, treating and / or reducing the risk of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer and / or gastric cancer, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from NPLVA (SEQ ID NO: 11), NPLVANPLVA (SEQ ID NO: 46) and / or FLAEDALNTV (SEQ ID NO: 126), or a fragment or variant thereof.

27. 27. The method of claim 26, wherein a recombinant phage and / or probiotic vaccine comprising FLAEDALNTV (SEQ ID NO: 126) is used to prevent, treat or reduce the risk of breast cancer.

28. 1. A method of preventing, treating and / or reducing the risk of multiple myeloma, comprising administering to a subject in need thereof: KRNIQFSCKNI (SEQ ID NO: 49), QTLEAQTLEA (SEQ ID NO: 58), and / or IQFSCIQFSC (SEQ ID NO: 59) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of:

29. 1. A method of preventing, treating and / or reducing the risk of cancer, comprising administering to a subject in need thereof a recombinant phage and / or a recombinant phage comprising at least one exogenous peptide epitope selected from the group consisting of ESFDGDPASNTAPLQPEQL (SEQ ID NO: 38), APKAQIKESLR (SEQ ID NO: 64), EDPAFYMLKGKLQYELQ (SEQ ID NO: 84) and / or SPRKLISVDSRSVSL (SEQ ID NO: 131), or a fragment or variant thereof. or administering a probiotic vaccine, wherein the cancer is selected from the group consisting of mucosal cancer, gastric cancer, melanoma, non-small cell lung cancer (NSCLC), head and neck squamous cell carcinoma (HNSCC), urothelial carcinoma, non-muscle invasive bladder cancer (NMIBC), colon or rectal cancer, esophageal cancer or certain gastroesophageal junction (GEJ) cancers, cervical cancer, renal cell carcinoma (RCC), advanced endometrial cancer, cutaneous squamous cell carcinoma (cSCC), and / or triple-negative breast cancer (TNBC).

30. 1. A method of preventing, treating and / or reducing the risk of mucosal melanoma, comprising administering to a subject in need thereof: ALLEIASCL (SEQ ID NO: 9), EDPAFYMLKGKLQYELQ (SEQ ID NO: 84), MYPPPY (SEQ ID NO: 10), MYPPPYMYPPPY (SEQ ID NO: 43), VELMYPPPYYLGIG (SEQ ID NO: 44), LMYPPPYYLGIGN (SEQ ID NO: 45), and / or SPRRKLISVDSRSVSL (SEQ ID NO: 131) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of:

31. 1. A method of preventing, treating and / or reducing the risk of prostate cancer, comprising administering to a subject in need thereof: ESKVDPSKAW (SEQ ID NO: 29), ESKVDPSK (SEQ ID NO: 12), MYPPPY (SEQ ID NO: 10), MYPPPYMYPPPY (SEQ ID NO: 43), VELMYPPPYYLGIG (SEQ ID NO: 44), LMYPPPYYLGIGN (SEQ ID NO: 45), ASPLLSGSQHFDVPPRQL (SEQ ID NO: 65), and / or KVIQAQTAFSANPANPAILS (SEQ ID NO: 67) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of:

32. 1. A method for preventing, treating, and / or reducing the risk of blood cancer, leukemia, chronic lymphocytic leukemia (CLL), small lymphocytic lymphoma (SLL), Waldenstrom's macroglobulinemia, chronic graft-versus-host disease, mantle cell lymphoma (MCL), marginal zone lymphoma, etc., comprising administering to a subject in need thereof: PTPEEDQILKK (SEQ ID NO: 63), DVIGRIENGERL (SEQ ID NO: 119), and / or RLIKEDVYL (SEQ ID NO: 120) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of:

33. A method for preventing, treating and / or reducing the risk of acute myeloid leukemia (AML), autoimmune and inflammatory diseases, canine cancer, etc., comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope corresponding to ASPLLSGSQHFDVPPRQL (SEQ ID NO: 65), or a fragment or variant thereof.

34. 1. A method for preventing, treating and / or reducing the risk of pulmonary fibrosis, comprising administering to a subject in need thereof: QYDSWPHYPE (SEQ ID NO: 66), YLSETVSNALGPQGRRLEE (SEQ ID NO: 73), EDPAFYMLKGKLQYELQ (SEQ ID NO: 84), MYPPPY (SEQ ID NO: 10), MYPPPYMYPPPY (SEQ ID NO: 43), VELMYPPPYYLGIG (SEQ ID NO: 44), LMYPPPYYLGIGN (SEQ ID NO: 45), and / or SPRRKLISVDSRSVSL (SEQ ID NO: 131) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of:

35. A method for preventing, treating, and / or reducing the risk of lung cancer, non-small cell lung cancer, etc., comprising administering to a subject in need thereof: SPRMSGLLSQTGYLSDNHILI (SEQ ID NO: 75), ASPLLSGSQHFDVPPRQL (SEQ ID NO: 65), SSDWVTLGTSASLLR (SEQ ID NO: 90), STPYVGLSLSCAANQ (SEQ ID NO: 91), PSPQYSWRINGIPQQHTQVLF (SEQ ID NO: 92), NPTTYQMDVNPEGKYS (SEQ ID NO: 95), HTPPLDPQELDILKT (SEQ ID NO: 96), RPLKGAPLPAL (SEQ ID NO: 101), EDPAFYMLKGKLQYELQ (SEQ ID NO: 84), MYPPPY (SEQ ID NO: 10), VVVGADVGKS (SEQ ID NO: 62), VVVGACVGKS (SEQ ID NO: 129), MYPPPYMYPPPY (SEQ ID NO: 43), VELMYPPPYYLGIG (SEQ ID NO: 44), LMYPPPYYLGIGN (SEQ ID NO: 45), GRADALRVLSK (SEQ ID NO: 122), LSRELSSLASQPLSR (SEQ ID NO: 123), and / or SPRRKLISVDSRSVSL (SEQ ID NO: 131) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of:

36. 1. A method for preventing, treating, and / or reducing the risk of thyroid eye disease, etc., comprising administering to a subject in need thereof: GRADALRVLSK (SEQ ID NO: 122), and / or LSRELSSLASQPLSR (SEQ ID NO: 123) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of:

37. 1. A method for preventing, treating and / or reducing the risk of cancer, comprising administering to a subject in need thereof: PIPHDGNLYPRLYPE (SEQ ID NO: 68), SKCRRCRLLD (SEQ ID NO: 69), QNTKCRCKPNFFC (SEQ ID NO: 70), TETEPPVAL (SEQ ID NO: 117), REALHEAL (SEQ ID NO: 118), VVVGADVGKS (SEQ ID NO: 62), VVVGACVGKS (SEQ ID NO: 129), and / or KTLETNPYVK (SEQ ID NO: 121) administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of: The method, wherein the cancer is selected from the group consisting of gastric cancer, melanoma, mucosal melanoma, lung cancer, non-small cell lung cancer (NSCLC), head and neck squamous cell carcinoma (HNSCC), urothelial carcinoma, non-muscle-invasive bladder cancer (NMIBC), colon or rectal cancer, esophageal cancer or certain gastroesophageal junction (GEJ) cancers, cervical cancer, renal cell carcinoma (RCC), advanced endometrial cancer, cutaneous squamous cell carcinoma (cSCC), pancreatic ductal carcinoma, and / or prostate cancer.

38. 38. The method of claim 37, wherein the recombinant phage and / or probiotic vaccine comprises KTLETNPYVK (SEQ ID NO: 121) and is used to prevent, treat or reduce the risk of colorectal cancer.

39. A method for preventing infection and / or disease manifestations of Korean fever virus, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of: KRNIQFSCKNI (SEQ ID NO: 49), QTLEAQTLEA (SEQ ID NO: 58) and / or IQFSCIQFSC (SEQ ID NO: 59), or a fragment or variant thereof.

40. 1. A method of preventing COVID-19 infection and / or disease manifestations, comprising administering to a subject in need thereof: AGICAS (SEQ ID NO: 14), AG-I-CAS (SEQ ID NO: 15; Ile="AUA" codon), VLYQDVNCT (SEQ ID NO: 19), QQQQGQTITKKSAAEASKKPRQKRTAVLVQDVGCTFSTVFPVTSFGPLVR (SEQ ID NO: 20), VLYQGVNCT (SEQ ID NO: 21), AGICASAGICAS (SEQ ID NO: 28), CVADYSVLY (SEQ ID NO: 33), HVGEIPVAY (SEQ ID NO: 34), NQKLIANQF (NQK-Q8) (SEQ ID NO: 35), NQKLIANAF (NQK-A8) (SEQ ID NO: 36), and / or RVAGDSGFAAY (SEQ ID NO: 37) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of:

41. A method for preventing, treating and / or reducing the risk of hepatitis C, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of CINGVCWTV (SEQ ID NO: 113) and / or KLVALGINAV (SEQ ID NO: 114), or a fragment or variant thereof.

42. A method for preventing, treating and / or reducing the risk of shingles, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of NLVPMVATV (SEQ ID NO: 115) and / or TPRVTGGGAM (SEQ ID NO: 116), or a fragment or variant thereof.

43. 1. A method for preventing, treating and / or reducing the risk of avian influenza infection and / or disease manifestations, comprising administering to a subject in need thereof: DAPF (SEQ ID NO: 22), KQIVERILEEE (SEQ ID NO: 23), RYLTDMTLEE (SEQ ID NO: 24), DDVKRENPEHEEYNN (SEQ ID NO: 25), RESGSS (SEQ ID NO: 26), DAPFKQIVERILEEE (SEQ ID NO: 30), RESGSSRYLTDMTLEE (SEQ ID NO: 31), EFINVPEGGS (SEQ ID NO: 39), EFINVPEEFINVPE (SEQ ID NO: 40), DAPPFDAPF (SEQ ID NO: 60), and / or RESGSSRESGSS (SEQ ID NO: 61) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of:

44. 1. A method of preventing, treating, and / or reducing the risk of rheumatoid arthritis, juvenile idiopathic arthritis, psoriatic arthritis, ankylosing spondylitis, Crohn's disease, ulcerative colitis, plaque psoriasis, hidradenitis suppurativa, or uveitis in a subject in need thereof: RSSSRTPS (SEQ ID NO: 13), VRSSSRTPSD (SEQ ID NO: 41), RSSSRTPSDK (SEQ ID NO: 42), ETPEGAEA (SEQ ID NO: 48), and / or ALDENTKLWYA (SEQ ID NO: 87) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of:

45. 1. A method of preventing, treating and / or reducing the risk of multiple sclerosis, comprising administering to a subject in need thereof: ESVDPKNYPKK (SEQ ID NO: 72), RPKSNIVLLS (SEQ ID NO: 97), RPKSNIVLLSAEEKKEQT (SEQ ID NO: 99), and / or SGPKPLFRRMSSLVGPTQ (SEQ ID NO: 98) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of:

46. 1. A method for preventing, treating and / or reducing the risk of osteoporosis, sclerosis, Van Buchem disease, osteonecrosis, osteoarthritis, rheumatoid arthritis, ankylosing spondylitis, bone tumors, multiple myeloma, bone metastasis, prostate cancer, glucocorticosteroid-induced osteoporosis, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of LPNAIGRGKWWRP (SEQ ID NO: 47) and / or CTHYMQLAC (SEQ ID NO: 127), or a fragment or variant thereof.

47. A method for preventing, treating and / or reducing the risk of primary hyperlipidemia, heterozygous familial hypercholesterolemia, hyperlipidemia, myocardial infarction, stroke and coronary revascularization, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope corresponding to REIEGRVMVTDPENVPEEDG (SEQ ID NO: 81), or a fragment or variant thereof.

48. A method for preventing, treating and / or reducing the risk of acne, signs of aging, wrinkles, fine lines, scars, hyperpigmentation, Kaposi's sarcoma lesions, melasma, psoriasis, and stretch marks, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope corresponding to GYYYRDDSLPVLLE (SEQ ID NO: 83), or a fragment or variant thereof.

49. A method for preventing, treating and / or reducing the risk of paroxysmal nocturnal hemoglobinuria, atypical hemolytic uremic syndrome, intravascular hemolysis, thrombosis, fatigue and the need for blood transfusions, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope corresponding to DKPVYTPDQSVKVRV (SEQ ID NO: 132) or a fragment or variant thereof.

50. 1. A method for preventing, treating and / or reducing the risk of asthma, severe asthma, atopic dermatitis, eczema, eosinophilic or oral steroid-dependent asthma, chronic rhinosinusitis with nasal polyposis, eosinophilic esophagitis, prurigo nodularis, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of NTVSCNRPHCITE (SEQ ID NO: 52), EERRR (SEQ ID NO: 53), EERRRREERRR (SEQ ID NO: 54), and / or AGAFAPSAAVA (SEQ ID NO: 82), or a fragment or variant thereof.

51. A method for preventing, treating and / or reducing the risk of asthma or severe eosinophilic asthma, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of (SEQ ID NO: 52), EERRR (SEQ ID NO: 53) and / or EERRRREERRR (SEQ ID NO: 54), or a fragment or variant thereof.

52. 1. A method of preventing, treating and / or reducing the risk of psoriasis, comprising administering to a subject in need thereof: LCTLAWAHPLVGHM (SEQ ID NO: 71), ALFDAQAQV (SEQ ID NO: 85), LLYDLTEKQVY (SEQ ID NO: 86), APNRTITVDDKMSLRLH (SEQ ID NO: 89), SLLPDSPVGQ (SEQ ID NO: 124), and / or SPSQPWQRL (SEQ ID NO: 125) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of:

53. 1. A method for preventing, treating, and / or reducing the risk of an inflammatory disease selected from the group consisting of psoriatic arthritis, Crohn's disease, ulcerative colitis, psoriasis, rheumatoid arthritis, atopic dermatitis, ankylosing spondylitis, hidradenitis suppurativa, and uveitis, comprising administering to a subject in need thereof: EIRQAGRPNKPDSI (SEQ ID NO: 57), and / or VLVESEHQV (SEQ ID NO: 88) The method comprises administering a recombinant phage or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of:

54. 1. A method for preventing, treating and / or reducing the risk of pain, comprising administering to a subject in need thereof: GLPSLGLTL (SEQ ID NO: 76), HRGEFS (SEQ ID NO: 77), HRGEFSHRGEFS (SEQ ID NO: 78), STLYREELMTTLLYYEIFL (SEQ ID NO: 93), and / or CEKNSVLEVIAYSSSSE (SEQ ID NO: 94) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of:

55. A method for preventing, treating and / or reducing the risk of chronic wasting, mad cow disease, comprising administering to a subject in need thereof a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope corresponding to DYEDRYYRE (SEQ ID NO: 100), or a fragment or variant thereof.

56. 1. A method for preventing, treating, and / or reducing the risk of a neurological disorder selected from the group consisting of Alzheimer's disease, Parkinson's disease, dementia, multiple sclerosis, epilepsy, Huntington's disease, muscular dystrophy, and migraine, comprising administering to a subject in need thereof: FRHDSGY (SEQ ID NO: 27), THLHWHTV (SEQ ID NO: 79), VQDYVQMKASELEQ (SEQ ID NO: 80), and / or INDPRLRNPSDKFIY (SEQ ID NO: 74) The method comprises administering a recombinant phage and / or probiotic vaccine comprising at least one exogenous peptide epitope selected from the group consisting of:

57. 59. The method of claim 58, wherein the disease is Alzheimer's disease.

58. 57. The method of claim 56, wherein the disease is migraine and the recombinant phage and / or probiotic vaccine comprises at least one exogenous peptide epitope selected from VQDYVQMKASELEQ (SEQ ID NO: 80).

59. 59. The method of any one of claims 23 to 58, wherein secretory IgA (SigA) is produced.

60. 60. The method of claim 59, wherein secretory IgA (SigA) is produced in an amount that is 2-100 fold, 2-90 fold, 2-80 fold, 2-70 fold, 2-60 fold, 2-50 fold, 2-40 fold, 2-30 fold, 2-20 fold, 2-10 fold, 2-9 fold, 2-8 fold, 2-7 fold, 2-6 fold, 2-5 fold, or 2-4 fold greater than the control when compared to the control.

61. The probiotic vaccine according to any one of claims 11 to 18, wherein the vaccine continuously produces lysogenic phages.

62. The probiotic phage according to any one of claims 11 to 18, or the recombinant phage according to any one of claims 12 to 18, wherein -TSGSGSGSGSGSGSG- is used as a linker between the coat protein and the exogenous peptide epitope.

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