FIELD OF THE INVENTION

The present invention generally relates to polypeptides which are immunomodulators and immunostimulants. Most specifically, the present invention relates to modified bacterial polypeptides having immunomodulatory and immunostimulatory activity. Yet more specifically, the modified polypeptides of the invention are the adjuvant components of a pharmaceutical composition for eliciting an immune response against the specific antigen present in the pharmaceutical composition. In a preferred embodiment, the invention provides a pharmaceutical composition for a vaccine which comprises an adjuvant component which is any one of the modified polypeptides according to the sequence listing of the present invention, one or more antigens, together with a pharmaceutically acceptable excipient.

STATE OF THE ART

In veterinary and medical sciences, immunomodulation is an area wherein extensive studies have been conducted to devise methods to improve disease resistance as well as to prevent or control immune disorders of host by optimum regulation of the immune system. Today, most infectious diseases of man and animals are treated and controlled mainly by using broad-spectrum antibiotics and vaccines. However, the antibacterial agents are becoming increasingly ineffective due to rapid emergence of resistant microbial strains. So, there is high requirement for novel and improved alternative therapeutic and prophylactic strategies to manage several diseases which are flaring at alarming pace because of the increase in international traffic, globalization and changing food habits. Immunomodulation is focused on manipulation of immune system to control the infections and other adverse health effects with precise regulation to avoid any complications while suppressive or potentiating efforts are made to benefit the animal and human health.

The immune system can be manipulated specifically by vaccination or non-specifically by immunomodulation.

It is generally accepted that vaccination is the most efficient and cost-effective form of preventing infectious diseases. Although most vaccines currently licensed are administered by parenteral route, this vaccination strategy usually fails to elicit adequate mucosal immune responses. Thus, diseases caused by mucosal pathogens are still among the major causes of death in developing countries. It is well documented that oral immunization can generate strong protective immunity at the intestinal mucosa as well as systemic (1-5). However, oral administration of antigens faces two major issues, one being degradation of the antigen by enzymes present at the gastrointestinal tract and the other being the induction of immune tolerance against the administered antigen (6-8). Current oral vaccines based on attenuated pathogens are usually capable of bypassing these difficulties but on the other hand present important safety concerns (9). The safe alternative consists on the development of oral vaccines based on killed pathogens or subunit thereof, but this holds major challenges since they are poorly immunogenic and usually require multiple doses and the use of an effective oral adjuvant (10, 11).

Two bacterial products are being used as oral adjuvants in the mouse model, cholera toxin (CT) fromV. choleraeand heat-labile enterotoxin (LT) fromE. coli(12). Since enterotoxicity seriously limits the practical use of these compounds in humans, modifications have been generated to reduce this effect. A modified version of CT lacking the A subunit (CTB) is now currently licensed as part of the Dukoral® vaccine for human use (13) and also a double mutant of LT (dmLT) which retains its adjuvant properties, is under clinical trial (14). However, neither of these molecules is capable of overcoming antigen degradation in the gastrointestinal tract. Interesting strategies to address this issue include antigen delivery through intestine-targeted pH-dependent microparticles (15), biodegradable nano or microparticles (16-18) antigen targeting to M cells (19) and protease inhibitors (20). However, there is still a need for appropriate adjuvants, and delivery systems that can be orally administered.

In addition, and as aforementioned, the immune system can also be manipulated non-specifically by immunomodulation. An immunomodulator may be defined as any biological or synthetic substance that can stimulate/suppress either innate or adaptive or both arms of the immune system. Immunomodulators include both immunostimulatory and immunosuppressive agents. Immunostimulatory agents are substances capable of enhancing host defense mechanisms to provide protection against infections. Their modes of actions include augmentation of anti-infectious immunity by the cells of the immune system, encompassing lymphocyte subsets, macrophages, dendritic cells and natural killer cells.

Dendritic cells (DC) represent a sentinel-like system with the capacity to produce cytokines and chemokines, capture and process antigens, to migrate into secondary lymphoid organs, and to activate naive T lymphocytes. They can be found in all lymphoid and most non-lymphoid tissues, including mucosal surfaces, like the lung and the gut, where intricate networks of DCs are situated to sense potential harmful exposures.

Dendritic cells (DC) are main gate-keepers of the immune system, bridging the innate and adaptive immune system. DCs are able to mature into inflammatory DCs at sites of inflammation in both autoimmune and allergic disease, thereby sustaining a continuous activation of the immune system at sites of inflammation. Maturation/activation of dendritic cells (DCs) is a key step in the induction of adaptive immune responses. The modulation and manipulation of DC functions through exposure of DCs to immunomodulatory agents is an approach that could result in therapeutic benefit in many clinical situations.

SUMMARY OF THE INVENTION

The present invention provides an immunomodulating polypeptide selected from the group of modified Ecotin fromSalmonella entericawithout signal peptide of SEQ ID NO. 1, modified aprin fromPseudomonas aeruginosawithout signal peptide, of SEQ ID NO. 2, modified staphostatin B fromStaphylococcus aureusof SEQ ID NO. 3, modified staphostatin A fromStaphylococcus aureusof SEQ ID NO. 4, and modified serine carboxypeptidase Y inh. fromHelicobacter pyloriof SEQ ID NO. 5.

Experimental evidence herein provided shows that each one of the polypeptides of the invention has proven to induce the activation of dendritic cells (DCs), thus being valuable immunomodulating agents.

The invention thus also concerns a pharmaceutical composition comprising (i) an immunomodulating polypeptide selected from the group of modified Ecotin fromSalmonella entericawithout signal peptide of SEQ ID NO. 1, modified aprin fromPseudomonas aeruginosawithout signal peptide, of SEQ ID NO. 2, modified staphostatin B fromStaphylococcus aureusof SEQ ID NO. 3, modified staphostatin A fromStaphylococcus aureusof SEQ ID NO. 4, and modified serine carboxypeptidase Y inh. fromHelicobacter pyloriof SEQ ID NO. 5, and (ii) a pharmaceutically acceptable excipient.

Experimental evidence shows that each one of the polypeptides of the invention has proven useful as an adjuvant for orally administered antigens. Administration of the polypeptides with an antigen results in the production of serum IgG and/or mucosal IgA against the antigen. Ag-specific CD4+and CD8+T cells increase also when polypeptides are delivered.

Accordingly, an object of this invention is an immunological adjuvant for vaccines, preferably administered via the oral route, where the adjuvant is a polypeptide selected from the group of modified Ecotin fromSalmonella entericawithout signal peptide of SEQ ID NO. 1, modified aprin fromPseudomonas aeruginosa, of SEQ ID NO. 2, modified staphostatin B fromStaphylococcus aureusof SEQ ID NO. 3, modified staphostatin A fromStaphylococcus aureusof SEQ ID NO. 4, and modified serine carboxypeptidase Y inh. fromHelicobacter pyloriof SEQ ID NO. 5.

A further object of this invention is a pharmaceutical composition for stimulating protective immunity at mucosal surfaces throughout the host.

An additional object of the invention is a pharmaceutical composition for inducing a protective immune response to any antigen, wherein the composition comprises at least an antigen in combination with one or more polypeptides selected from the group of modified Ecotin fromSalmonella entericawithout signal peptide of SEQ ID NO. 1, modified aprin fromPseudomonas aeruginosawithout signal peptide, of SEQ ID NO. 2, modified staphostatin B fromStaphylococcus aureusof SEQ ID NO. 3, modified staphostatin A fromStaphylococcus aureusof SEQ ID NO. 4, and modified serine carboxypeptidase Y inh. fromHelicobacter pyloriof SEQ ID NO. 5.

According to another embodiment, the composition of the invention is a component of a vaccine against any specific antigen where a specific antibody or cellular response would be useful in active acquired immunity to a particular disease.

According to another embodiment, the composition of the invention is a component of a vaccine or formulation composed of any specific antigen to which the induction of a specific immune response is required or desired.

Accordingly, the present invention provides a vaccine or formulation comprising at least an antigen and a polypeptide selected from the group of modified Ecotin fromSalmonella entericawithout signal peptide of SEQ ID NO. 1, modified aprin fromPseudomonas aeruginosawithout signal peptide, of SEQ ID NO. 2, modified staphostatin B fromStaphylococcus aureusof SEQ ID NO. 3, modified staphostatin A fromStaphylococcus aureusSEQ ID NO. 4, and modified serine carboxypeptidase Y inh. fromHelicobacter pyloriof SEQ ID NO. 5, and optionally a pharmaceutically acceptable vehicle.

According to another object of the present invention, the nucleotide sequences encoding the polypeptides of the invention are also provided. Said nucleic acid sequences are selected from SEQ ID NO. 6 to 10, wherein SEQ ID NO. 6 codifies for the modified Ecotin fromSalmonella(without signal peptide); SEQ ID NO. 7 codifies for the modified aprin fromPseudomonas aeruginosa(without signal peptide); SEQ ID NO. 8 codifies for the modified staphostatin B fromStaphylococcus aureus; SEQ ID NO. 9 codifies for the modified staphostatin A fromStaphylococcus aureus; and SEQ ID NO. 10 codifies for the modified serine carboxypeptidase Y inhibitor fromHelicobacter pylori.

According to another object of the present invention, a method is provided for stimulating innate immune response in a subject, comprising administering to said subject an effective amount of the pharmaceutical composition comprising one or more polypeptides of the invention.

According to another object of the present invention, a method is provided for enhancing the immune response against an antigen in a subject, comprising administering to said subject an effective amount of the pharmaceutical composition or of a vaccine comprising one or more polypeptides of the invention.

Another object of the invention is a method for inducing protective (sometimes termed adaptive) immunity to an antigen by administering to a subject in need thereof at least one dose of an effective amount of an antigen and an effective amount of one or more of the polypeptides of the invention.

In particular, another object of the invention is a method for inducing protective (sometimes termed adaptive) immunity to pathogen-derived antigens by administering to a subject in need thereof at least one dose of an effective amount of a pathogen-derived antigen and an effective amount of a polypeptide selected from the group of modified Ecotin fromSalmonella entericawithout signal peptide of SEQ ID NO. 1, modified aprin fromPseudomonas aeruginosawithout signal peptide, of SEQ ID NO. 2, modified staphostatin B fromStaphylococcus aureusof SEQ ID NO. 3, modified staphostatin A fromStaphylococcus aureusof SEQ ID NO. 4, and modified serine carboxypeptidase Y inh. fromHelicobacter pyloriof SEQ ID NO. 5, in an oral or parenteral acceptable pharmaceutical composition.

The antigen may consist of a peptide, protein, polysaccharide or any molecule or chemical moiety component of an organism or of a non-living organism or extract of said organism, to which the host may induce an immune response. Alternatively, the antigen may consist of substances containing epitopes shared by substances (allergens) to which the host has previously established an allergic immune response evoking allergic manifestations.

According to a preferred embodiment, the antigen is selected from the group of pathogen-derived antigens, tumor cell antigens, self-antigens and allergens. Pathogen-derived antigens may be from bacteria, virus, parasites or fungi.

According to a preferred embodiment, the pathogen-derived antigen is selected from the group of viral antigens, bacterial antigens, parasitic antigens and fungal antigens.

According to another embodiment, the antigen is an antigen of a virus selected from the group comprising, without limitation, Poxviridae, Herpesviridae, Herpes Simplex virus 1, Herpes Simplex virus 2, Adenoviridae, Papovaviridae, Enteroviridae, Picornaviridae, Papillomaviridae Parvoviridae, Reoviridae, retroviridae, influenza viruses, parainfluenza viruses, mumps, measles, respiratory syncytial virus, rubella, Arboviridae, Rhabdoviridae, Arenaviridae, Hepatitis A virus, Hepatitis B virus, Hepatitis C virus, Hepatitis E virus, Non-A/Non-B Hepatitis virus, Rhinoviridae, Coronaviridae, Rotaviridae, Vaccinia virus.

According to another embodiment, the antigen is an antigen of a bacterium selected from the group comprising, without limitationStreptococcus pyogenes, Streptococcus pneumoniae, Neisseria gonorrheae, Neisseria meningitidis, Corynebacterium diphtherias, Clostridium botulinum, Clostridium perfringens, Clostridium tetani, Hemophilus influenzae, Klebsiella pheumoniae, Klebsiella ozaenae, Klebsiella rhinoscleromatis, Staphylococcus aureus, Vibrio cholerae, Escherichia coli, Pseudomonas aeruginosa, Campylobacter(Vibrio) fetus,Aeromonas hydrophila, Bacillus cereus, Edwardsiella tarda, Yersinia enterocolitica, Yersinia pestis, Yersinia pseudotuberculosis, Shigella dysenteriae, Shigella flexneri, Shigella sonnei, Salmonella typhimurium, Treponema pallidum, Treponema pertenue, Treponema carateneum, Borrelia vincentii, Borrelia burgdorferi, Leptospira icterohemorrhagiae, Mycobacterium tuberculosis, Francisella tularensis, Brucella abortus, Brucella suis, Brucella melitensis, Mycoplasmaspp.,Rickettsia prowazeki, Rickettsia tsutsugamushi, Chlamydiaspp,Listeria monocytogenes.

According to another embodiment, the antigen is an antigen of a protozoan selected from the group comprising, without limitation Entomoebahistolytica, Trichomonas tenas, Trichomonas hominis, Trichomonas vaginalis, Trypanosoma gambiense, Trypanosoma rhodesiense, Trypanosoma cruzi, Leishamania donovani, Leishamania tropica, Leishmania braziliensis, Pneumocystis pneumonia, Plasmodium vivax, Plasmodium falciparum, andPlasmodiummalaria.

According to another embodiment, the antigen is an antigen of a Helminth selected from the group comprising, without limitationEnterobius vermicularis, Trichuris trichiura, Ascaris lumbricoides, Trichinella spiralis, Strongyloides stercoralis, Schistosoma japonicum, Schistosoma mansoni, Schistosoma haematobium, and hookworm.

According to another embodiment, the antigen is an antigen of a fungus selected from the group comprising, without limitationCoccidioides immitis, Aspergillus fumigatus, Candida albicans, Blastomyces dermatitidis, Cryptococcus neoformans, andHistoplasma capsulatum.

According to another embodiment, the antigen is a tumor-derived antigen selected from the group including, for example, without limitation AFP (alphafetoprotein), ETA (epithelial tumor antigen), CEA (carcinoembryonic antigen), MAGE (melanoma-associated antigen), MUC-1, abnormal products of ras, p53, TPBG (trophoblast glycoprotein), CEA (colorectal carcinoma), Immature laminin receptor (RCC), TAG-72 (prostate carcinoma), HPV E6, E7 (Cervical carcinoma), BING-4 (Melanoma), Calcium-activated chloride channel 2 (Lung carcinoma), Cyclin-B1(Multi), 9D7 (RCC), Ep-CAM (Breast carcinoma), EphA3 (Multi), Her2/neu (multi), Telomerase (multi), Mesothelin (ductal pancreatic carcinoma), SAP-1 (colorectal carcinoma), surviving (multi), BAGE family (multi), CAGE family (multi), GAGE family (multi), MAGE family (multi), SAGE family (multi), XAGE family (multi), NY-ESO-1/LAGE-1 (multi), PRAME (multi), SSX-2 (melanoma, multi), Melan-A/MART-1 (melanoma), Gp100/pme117 (melanoma), Tyrosinase (melanoma), TRP-1/2 (melanoma), P. polypeptide (melanoma), MC1R (melanoma), Prostate-specific antigen (PSA, prostate), β-catenin (melanoma, prostate, HCC), BRCA1/2 (breast, ovarian carcinoma), CDK4 (multi), CML66 (CML), Fibronectin (multi), MART-2 (melanoma), p53 (multi), Ras (multi), TGF-βRII (colorectal carcinoma), MUC1 (ductal carcinoma, RCC), Ig, TCR (B, T leukemia, lymphoma, myeloma).

According to another embodiment, the antigen is a protective antigen. Protective antigens are specifically targeted by the acquired immune response of the host and able to induce protection in the host against infectious and non-infectious diseases. Examples of protective antigens for different diseases—without limitation—are: Lef; LF; PA; PA63; PA63-LAMP1; PA83; Pag; PagA; TPA-PA63; TPA-PA63-LAMP1; UQ-PA63; BoNT/A; BoNT/A1; BoNT/B; BoNT/C(Hc50); BotA; FHc; Fragment C fromClostridium tetani; TTC; Tetanus toxoid; Tox; Diphteria toxoid; spaA; Hly; p60; Ag85B; Ag85A; Ag85A; Ag85B; Apa; bfrA; DnaK; ESAT-6; EsxA (ESAT-6); EsxB; FbpA (Ag85A); FbpB; FbpD; HBHA; KatG; Mpt63; MPT64; MPT83; Mtb72F; PE20; PepA; PPE14; PPE18; PPE31; PstS1; PstS3; Rv1806; Rv1807; Rv2660c; ClfA; can; efb; FnbA; isdB; MecA; BipA; BPS; Rib; Sip; binding protein Eag; collagen binding, ancillary pilus subunit Cne; Fbp; Fnz; HAP; Sfs; CbpA; ClpP; GltX; Gnd; Lp1A; phpA-79; PiaA; Ply; PsaA; PspA; psrP; SrtA; emm1; FbaA; FBP54; Sfb; Sib35; APP7_1016; ApxIA; apxIA; ApxIIA; apxIIA; CP; Lipopolysaccharide; AroA; AsaP1; brkA; Cpn60; CyaA; FhaB; Prn; PtxA; PtxB; PtxC; PtxD; PtxE; S1; DbpA; OspA; OspB; OspC; AsnC; L7/L12; P39; BAB1_0278; BCSP31; Bfr; Bp26; cob; DnaK; Gap; IalB; L7/L12; Omp16; Omp19; Omp25; Omp31; ORF; P39; Rp1L; SodC; SurA; Tig; FliC; LolC/E; LPS; PotF; CadF; CjaA fromC. jejuni; CjaA fromC. jejuniRM 1221; FlaA fromC. jejuni81-176; FlaA fromC. jejuniNCTC 11168; FlaC; FspA1; FspA2; Peb1A; PorA; MOMP; PmpE/F-2; PmpG-1; Rp1F; Tarp; TC0439 adherence factor; TC0512 outer membrane protein, putative; TC0757 hypothetical protein; TC0767 hypothetical protein; TC0768 hypothetical protein; ompA; CAB049 putative penicillin-binding protein; CAB613 putative peptidase; DnaK; DnaX; GatA; GatB; GatC; Om1A; Omp1; Pomp90A; CopN; FabD; HSP-60; LcrE; PknD; Ssb; MOMP; OmpA; HspB; P1; esa1; et18; eta6; fliC; C0393; C3389; C4424; CS1; Eae; ECSE_P3-0034; EltB; FimH; FyuA; Hma; IreA; IroN; Iss; IutA; StxB2; Tir; FopA; FopB; GroEL; Ig1C1; Ig1C2; LpnA; LPS; TUL4; Wbt1; D15; Hap; Hpd; HtrA; NucA; Omp26; OmpP1; OmpP2; OmpP5; P145; P4; Pal; Tbp2; gapA; Catalase; GltA; Hsp60; HspA; HspB; NAP; oipA; UreB; VacA; ligB; ompL1; TM-1; P97; ExbB; GNA1030; GNA1162; GNA1220; GNA1870; GNA1946; GNA2001; GNA2091; GNA2132; GNA33; GNA992 (hsf); gp120; LctP; LpdA; LPS; NadA; NspA; MC58; P1; TBP2; TbpA; OmpA; ompH; ompH; OprF; OprI; PcrV; pilA; OmpA; OmpB; SpaP; DnaJ; GroEL; Hs1U; IroN; SopB; TviB; GuaB; IpaB; IpaC; IpaD; Spa32; GlpQ; TmpB; Tp92; OmpU; CtxB; TcpA; TcpF; LcrV; Caf1; HpmB; HtpG; Irp7; LcrV; Lo1A; Pla; Psn; V antigen; YapF; YdeN; YerA; YopD; YopE; YopH; YopO; YPMT1.84 (F1 capsule antigen); YP00420; YP00612; YscF; pYV0055; Fusion protein (NCBI protein GI 6018112); G from BRSV; gB; gIV; UL44; UL49; US6; US7; Env/Rex/Tax; F gene; E2; VP1; VP2; E0; Gp55; E2; 18H6 synthetic E protein; E protein; E protein from Dengue 1; E protein from Dengue 3; E protein from Dengue 4; M protein; NS1; PrM; prM from Dengue 1; prM from Dengue 2; prM from Dengue 3; VP1; EBOV NP; GP from Reston ebolavirus; GP from Sudan ebolavirus; GP from Zaire ebolavirus; NP; SGP; VP24 from Reston ebolavirus; VP24 from Zaire ebolavirus; VP30 from Reston ebolavirus; VP30 from Zaire ebolavirus; VP35 from Reston ebolavirus; VP35 from Zaire ebolavirus; VP40 from Reston ebolavirus; VP40 from Zaire ebolavirus; ZGP; env from FeLV-A/Glasgow; Env from strain FRA; Gag; 1D; 2A; 3C; 3D; FMDV A12 genome; P1; VP1; gB; UL27; ANDVsSgp1; G1; G1 and G2; Gc; HTNVsSgp1; M Segment; N protein [Topografov virus]; PUUVsSgp1; S nucleocapsid protein from Dobrava-Belgrade virus; SNVsSgp1; G glycoprotein; HAVgp2; HBVgp2 pre-S1/pre-S2/S; Hepatitis B surface antigen; Large S protein; PreS2 middle surface protein; PreS2+S (Middle protein); S protein from Duck Hepatitis B virus; Small envelope antigen; Small S protein; p27; gB from HSV-1 KOS; ICP27 from HSV-1; ICP27 from HSV-2; UL27 from HSV-1; UL27 from HSV-2; UL44 from HSV-1; UL44 from HSV-2; US4; US5; US6; US7; US8; G glycoprotein; N nucleocapsid protein; VP1; Glycoprotein B; pp65; Env; env from clade B; env from HIV 2; env from HIV-1 isolate 037 clone 08; env from HIV-1 isolate 715; env from HIV-1 strain 96ZM651; env from HIV-1 strain 976 clone 17; env from HIV-1 strain Ba-L; env from SHIV-89.6; env from SHIV89.6P; env from SIV; Env Gp160; env MN isolate; Envelope polyprotein from HXB2; gag from clade A; Gag from HIV 1; gag from HIV 2; gag from HIV subtype B; gag from HIV-1 vector pNL4-3; gag from HXB2; gag from SHIV-89.6; Gag protein from SIV-mnd 2; gag-pol; gag-pol SIV; gp120; gp120 from MN; gp120 Indian subtype C; gp41; Nef; nef from HIV 2; pol from HIV 1; pol from SHIV-89.6; rev from HIV 1; rev from SHIV-89.6; Tat; tat from HIV 2; tat from SHIV-89.6; vif from SHIV-89.6; vpr from SHIV-89.6; vpu from SHIV-89.6; Calreticulin from humans; E6; E7; L1; L2; F protein; G protein; M membrane protein; N protein; S1; VP2; VP2-VP4-VP3; G glycoprotein; ORF; VP2; HA from influenza A virus, NA from Influenza A virus, NP from Influenza A virus (A/chicken/Henan/12/2004(H5N1)); NP from Influenza A virus, NP from Influenza B virus; Envelope protein; M protein; NS1; NS3; NS5; PreM; PrM; GPC; LASVsSgp1 nucleoprotein; LASVsSgp2 glycoprotein; NP; GP; GP from Lake Victoria marburgvirus Musoke; GP from Zaire ebolavirus; NP; NP from Lake Victoria marburgvirus; NP from Zaire ebolavirus; NP Marburgvirus—Ravn; SGP; VP35; MDV040; MDV095; F fusion protein; H hemagglutinin protein; M matrix protein; N nucleocapsid protein; P phosphoprotein; HN; gM; gN; F fusion protein; F protein from strain D26; HN hemagglutinin-neuraminidase; F fusion protein; G glycoprotein; VP4; ORF2; GP5 from CH-1a; GP5 from YA1; ORF7; ubiquitin; GB; GI; GII; GIII; Gp50; IE180; UL44; US6; VP60; env; gag; VP22; RVFVsMgp1 M protein; RVFVsSgp2 N protein; RPVgp5 F; RPVgp6 H; VP2; VP4; VP6; VP7; S protein; env; E2 from Sindbis virus; TBEVgp1 polyprotein; A27L; A33R; A34R; A36R; A4L; B5R; DBL; HSR; UR; ORF67; ORF68; A21L; H3L; UR from Monkeypox virus Zaire-96-I-16; 26S mRNA; C-E3-E2-E1-6K; E1 glycoprotein; E2 envelope protein; Glycoprotein; nucleocapsid; E protein; PrM; 26S; 6K; E1; E2; E (envelope glycoprotein); NS1 (non-structural protein 1); 11C5; 12D3; 21B4; RAP-1; CP15/60; CP23; gam56; Gam82; 3-1E; 3-1E; GX3262; MIC2; Rho; TA4; CEL-170/4; Gal/GalNAc lectin; GEh29; SREHP; CWP2; CP cysteine proteinase; Gp46; M2; P4; WD; A2; Beta-tubulin; BT1; F1-ATPase; GRP-78; HASPB1; Hsp70; KMP-11; MspC; NH; ORFF; P1; H1; HASPB1; LIPO-A; P36/LACK; SMT; Cpa; Cpb; Gp63; H2B; KMP-11; LACK; LmSTI1; PSA-2; SP; SW3.1; TSA; MIC10; NcDG1; NcDG2; NcMAG1; NcMIC1; NcPDI; NcSAG1; NcSRS2; ABRA; AMA-1 from (P. berghei; P. chabaudi;P. falciparum3D7 orP. knowlesi); CS fromP. bergheistr. ANKA; CS fromP. falciparum; CS fromP. yoeliistr. 17XNL; CSP fromP. knowlesi; eba-175; EMP1; GLURP; HEP17; HSP60 fromP. yoelii; Hsp90; Kbeta; LSA-3; MSP-1 fromP. falciparum; MSP1 fromP. berghei; MSP1 fromP. knowlesi; MSP1 fromP. vivax; MSP1 fromP. yoeliistr. 17XNL; MSP3; MSP4/5; MSP8; PF10_0155 enolase; pfCelTOS; Pvs25; RESA; SERA-5; SSP2; SSP2 fromP. knowlesi; TRAP fromP. falciparum; SAG1; FABP; II12a; Paramyosin; Sj23; TPI; calpain; GPX; Sm23; SOD; Spag-1; TA17050 merozoite-piroplasm surface antigen Tams1; P67; GRA1; GRA4; hsp70; MIC13; MIC3; MICS; Rop2; ROPE; SAG1; SAG2; MAPP15; TSA; amastigote surface protein-2; ASP-2; ASP-2 from strain Tulahuen; ASP-2 from the Y strain; ASP1; CRP-10; Cruzipain; G2; Par1; protein G4; Tc00.1047053434931.10; Tc24 fromTrypanosoma cruzi; TS; TS from strain Y; TSA-1; AMN1; ELI-Ag1; GEL1; PEP1; PLB; Pmp1; Para; TcrP; URE; Angpt1; Cd276; Cd86; Fas; Flt3; Il18; Il25; Il33; LPS lipopolysaccharide; Ph1 p 12 (Phleum pratense); Ph1 p I allergen [Phleum pratense]; Ph1 p7 protein [Phleum pratense]; Tbx21; Tnfrsf13b; Tnfrsf9; COL2A1; Collagen type II; hsp65 fromM. leprae; ACPP; AFP; AFP from human; ANTXR1; Cc121a; Cd401g; Cd80; Cd86; CEA; CEACAM3; CTAG1B; CTLA4; E2A; Eng; EPCAM; ERBB2; Fap; Fas1; FASLG; Flt3; Fosl1; Fragment C from tetanus toxin; GAST; GM-CSF; gp100; gp75; GPC3; GRP; H2-K1; Heat shock protein; hsp70; HuD; IgG Fc; Il18; Immunoglobulin; KDR; KLK3; MAGE-1; MAGEA3; Mcam; MLANA; MUC1; Muc1; PAP; Pdgfrb; Pme117; Prostate specific antigen; Prostate stem cell antigen; Prostate-specific membrane antigen; PSCA; SCFV; SCGB2A2; SILV; SLC5A5; Steap1; Surface antigen from Hepatitis B; Survivin; TERT; TNFRSF18; TPBG; TRP-1; TRP-2; Trp2; Trp53; Tyrosinase; Tyrosinase from Mouse; TYRP1; Ubiquitin; VEGFA; WT1; BAX; Gad65; IgG Fc; MBP from guinea pig; Mog; IRBP from humans; Ricin A chain.

According to another embodiment, the antigen comprises antigenic material derived from heat-killed/formalin killed whole organisms or extracts of organisms causing diseases such as the ones listed above without limitation.

Additionally, the present invention provides experimental data showing that the Ecotin polypeptide is a good antigen againstSalmonellainfection in mice.

It is apparent to someone who is skilled in the art that this invention will be useful for any specific antigen where a specific neutralizing antibody response or specific cellular immune response would be useful for eliminating the physiological or disease state associated with that antigen.

