CN106478802B - TNF-α protein B cell epitope, multiple antigenic peptide and application containing this epitope - Google Patents

Abstract

The present invention provides a kind of B cell epitope of TNF-α albumen and the multiple antigenic peptide based on the epitope, the 8 branched peptide conformations synthesized in conjunction with MAP scheme, and the application in preparation ulcerative colitis vaccine.The immunological investigation field that MAP design scheme is applied to small molecule autoantigen is creatively filtered out into the B cell Dominant Epitopes of humanTNF-α's albumen by a large number of experiments.On this basis, using MAP design scheme, obtain the 8 branched peptide conformations using lysine as core matrix, which belongs to autoantigen epitope vaccine, thus have convenient drug administration and number it is few, only need to inject on a small quantity can induce generate antibody the advantages of；And have safety and tolerance are good, prevention and treatment is simultaneous the advantages such as to apply, it is expected to provide important theoretical basis for the research and development of self-antigen MAP vaccine, new thinking can also be provided for clinical treatment ulcerative colitis.

Description

TNF-α protein B cell epitope, multiple antigenic peptide and application containing this epitope

Technical field

The invention belongs to immunology and disease for digest fields, and in particular to a kind of the B cell epitope and base of TNF-α albumen In the multiple antigenic peptide of the epitope, the 8 branched peptide conformations synthesized in conjunction with MAP scheme, and in preparation ulcerative colitis vaccine Application.

Background technique

TNF-α be it is a kind of there is the active cell factor of complex biological, in addition to can in vivo, outside outer killing tumor cell, Also there is induction inflammatory reaction, viral infection resisting, adjust immunity of organism, promote the multiple functions such as cell Proliferation and activation.However Studies have shown that when TNF-α overexpression, it, which can be cooperateed with, generates a variety of pathology damages, as inflammatory bowel disease, infectious diseases, Dyscrasia etc.；Meanwhile TNF-α is also a kind of human body autoantigen that can be used as therapy target, neutralizes internal excessive TNF-α then Above-mentioned pathologic damage can be made to mitigate.There are mainly two types of the immunotherapeutics for being directed to this anthropoid autoantigen at present: First method is the passive immunity that internal autoantigen is directly neutralized to patient injection monoclonal antibody: such as anti-tnf-alpha Dan Ke Grand antibody (infliximab, infliximab), is most common passive immunotherapy biological agent.Infliximab It is a kind of chimera monoclonal anti-TNF-alpha antibody of synthesis, is made of 75% humanized and 25% mouse, it can be with monokaryon macrophage Cell and the T cell film mating type TNF-α of activation combine or in conjunction with the TNF-α dissociated in blood plasma, and are neutralized, to reach To the effect for reducing TNF-α.Now think, infliximab can be used for reactionless to hormone and immunosuppressor or not be resistant to And be not suitable for treatment be treated surgically, severe activity Crohn disease (Crohn disease, CD) patient；It is clinical Test shows that it also has treatment ulcerative colitis (Ulcerative colitis, UC) curative effect more certainly.But The defects of that there are dosages is big by Infliximab, high production cost, need to be repeatedly used for a long, Yi Yinqi hypersensitivity；In addition, needle To defect existing for such monoclonal antibodies drug, Nature magazine also once delivered entitled " whether super antibody drug can quilt Tame? " article, it is indicated that potential hazard that such drug-induced immune response generates, which has become, limits its pass used Key factor.Second of immunotherapeutic is to make to generate in vivo to the immune autovaccine with auto-antigen epitope of patient to be directed to The active immunity of the polyclonal antibody of autoinducer molecule: in view of vaccine have production cost it is low, it is easy to use, only need to inject on a small quantity just The advantages that generating antibody and the defect of antibody and soluble recepter drug can be overcome can be induced, therefore, by passively receiving list Clonal antibody drug diversion seeks the active immunity vaccine (i.e. autovaccine) for mankind itself's albumen, it has also become anti-from status The new developing direction of sub- immunization therapy.

The research for surely belonging to epiposition vaccine most popular at present of autovaccine research field.Epiposition vaccine includes t cell epitope Vaccine and B cell epiposition vaccine.The research and development of epiposition vaccine can provide a better treatment means for a variety of diseases, but by Belong to autoantigen in autoinducer molecule, molecular weight is smaller, structure is single, often results in immunogenicity reduction, cannot lure in vivo The ideal immune response of hair, especially humoral immune reaction, this is also that B cell epiposition vaccine studies relatively small number of main original Cause.In order to solve this problem, small peptide vaccine mostly uses the method for epitope polypeptide crosslinking carrier protein to improve its immunogenicity, But since carrier protein itself is also macromolecular exotic antigen, the antibody often induced is mostly to be directed to carrier protein rather than purpose table Position.

In recent years in the research field of tumor associated antigen (tumor associated antigen, TAA), there is scholar to mention Multiple antigenic peptide (muptile antigenic peptides, MAP) design scheme is gone out, and immunogenicity small using molecular weight is weak Lysine (K) be core matrix, the identical or different monomeric peptide of several (generally 4 or 8) epitopes is coupled Together, dendritic structure is formed.This design pattern can not only simulate natural epitopes conformation very well, and no longer need to coupling and carry Body protein just can activate humoral immunity, induce high titre, high-affinity antibody.Therefore, using the branched of MAP conceptual design More epitope complex peptides are expected to the defect for improving the quality of peptide vaccine, removing carrier protein.

MAP design scheme is mainly used for constructing the T cell of the macromoleculars antigens such as tumor associated antigen and B cell epitope is mostly anti- The research of former peptide has more application in the research of heparinase.Since autoantigen is relatively with macromolecular antigen, there is day Right immune tolerance, molecular weight is smaller, epitope peptide is shorter and is more difficult to screen, be less susceptible to the defects of causing humoral immune reaction, therefore, MAP design scheme was not applied to itself small molecule antigens by this field researcher.However, we are based in the past to B cell The numerous studies basis of epitope MAP vaccine, early period under the subsidy of state natural sciences fund, successfully use MAP scheme constructs With more strongly immunogenic TNF-α B cell epiposition vaccine.

Summary of the invention

In view of the above-mentioned deficiencies in the prior art, it is an object of the present invention to provide the B cell epitope and base of a kind of TNF-α albumen In the epitope, in conjunction with the multiple antigenic peptide for the 8 branched peptide conformations that MAP scheme synthesizes, and in preparation ulcerative colitis vaccine Application.

The purpose of the present invention is what is be achieved through the following technical solutions: a kind of B cell epitope of TNF-α albumen, the table The amino acid sequence of position is VMVPRRKAGTSKS or VCFTLNKSQSNQE.Dominant Epitopes are VCFTLNKSQSNQE.

A kind of ulcerative colitis multiple antigenic peptide, the multiple antigenic peptide are to design by MAP, and immunogene small with molecular weight Property weak lysine (K) be core matrix, be coupled together with epitope polypeptide that sequence is VCFTLNKSQSNQE, form 8 branches The multiple antigenic peptide of peptide conformation.

A kind of B cell epitope for humanTNF-α's albumen that amino acid sequence is VMVPRRKAGTSKS or VCFTLNKSQSNQE Application in preparation ulcerative colitis vaccine.

A kind of application of ulcerative colitis multiple antigenic peptide in preparation ulcerative colitis vaccine.

The beneficial effects of the present invention are: MAP design scheme is creatively applied to small molecule autoantigen by the present invention Immunological investigation field predict, screen, identifying with good hydrophily, accessibility, plasticity by a large number of experiments, And it is located at the B cell Dominant Epitopes of albumen stretched out structure or humanTNF-α's albumen in random coil structure in secondary structure. On this basis, using MAP design scheme, the 8 branched peptide conformations using lysine as core matrix are obtained, the 8 branched peptide conformation Polypeptide vaccine compared with the existing technology, has the advantage that

TNF-α B cell epitope MAP vaccine belongs to autoantigen epitope vaccine, thus have convenient drug administration and number it is few, only Need to inject on a small quantity can induce the advantages of generating antibody；And have relative low price, safety and tolerance it is good, prevention and treatment it is simultaneous The advantages such as apply.Therefore, it if the exacerbation and repeatedly of the ulcerative colitis state of an illness can be prevented and treated by TNF-α MAP vaccine, is bound to mention High patients of ulcerative colitis compliance is improved the quality of living and Economy type medicine expense, while also having agreed with new medical reform " prevention is The thinking of master " and the theory of Xi Jinping General Secretary's " Health China ", are expected to provide for the research and development of self-antigen MAP vaccine important Theoretical basis can also provide new thinking for clinical treatment ulcerative colitis.

Detailed description of the invention

Fig. 1 is humanTNF-α's B cell epitope DNAstar prediction result figure；

Fig. 2 is humanTNF-α's B cell epitope Bcepred prediction result figure；

Fig. 3 is 8 candidate branched peptide MAP structural schematic diagrams；

Fig. 4 is MAP1 and MAP2 and the binding force schematic diagram that TNF-α antibody is commercialized；

Fig. 5 is that Western blotting detects each MAP antibody specificity result figure；

Fig. 6 is MAP immunized mice serum specific antibody titre dynamic change figure；

Fig. 7 is that MAP antibody weakens TNF-α to the lethal effect schematic diagram of L929 cell；

Fig. 8 is the mouse Colon tissue MPO activity schematic diagram for being vaccinated with MAP vaccine, shows that MPO activity is substantially reduced；

Fig. 9 is inoculation MAP vaccine mouse Colon Histopathologic damage plan, is shown after being inoculated with MAP vaccine, pathologic It damages substantially reduced；

Figure 10 is that inoculation MAP vaccine mouse Colon tissue CAM 120/80 expresses schematic diagram, is shown after being inoculated with MAP vaccine, CAM 120/80 expression significantly reduces.

Specific embodiment

Embodiment 1: the screening of humanTNF-α's B cell epitope Dominant Epitopes and the building of multiple antigenic peptide.

1. retrieving NCBI GenBank database, humanTNF-α's protein amino acid sequence is obtained；Using DNAStar, Antivirus design tool Bcepred on GOLDENKEY (Military Medical Science Institute) software and internet, analysis obtain parent The strongest amino acid sequence information of antigenicity and B cell epitope in pool；According to Epitope prediction, selection is rich in hydrophilic ammonia Base acid length is about the peptide fragment of 6-25 amino acid, while avoiding the hydrophobic amino acid residues of 4 or more continuous adjacents and containing 2 With the peptide fragment of cysteine, in order to avoid pass through the polymer of disulfide bond formation complexity.

2. analyzing the prediction result of software (Fig. 1) and internet design tool (Fig. 2).With 13 amino acid residues for one Group analyzes its hydrophily, accessibility, plasticity and the antigenicity in antigenic index assessment humanTNF-α's molecule high-hydrophilic region. Several analysis methods are finally integrated, the overlapping region of each result is taken, determine 22-34 (VMVPRRKAGTSKS), 54-66 (VCFTLNKSQSNQE) peptide fragment composed by amino acid has good hydrophily, accessibility and plasticity, in secondary structure It is the B cell candidate's epi-position of TNF-α albumen in albumen stretched out structure or random coil structure.

3. using MAP design scheme, entrusting Zhongtai Bio-Chem. Co., Ltd., Hangzhou by the with lysine (K) for core matrix 22-34 (VMVPRRKAGTSKS), 54-66 (VCFTLNKSQSNQE) amino acid C-terminal be coupled at core with 8 branched peptide conformations On the amino of matrix lysine, isolated and purified by HPLC, Mass Spectrometric Identification analysis and peptide fragment amino acid sequencing, prepare TNF- respectively α B cell candidate's epi-position MAP polypeptide, and it is respectively designated as MAP1 and MAP2, MAP1, MAP2 structural schematic diagram are shown in Fig. 3.

4. the screening of candidate MAP is as follows:

(1) binding ability of the MAP antibody from MAP in vitro: 1) the different MAP for being respectively 10 μ g/mL with concentration, dilution For the carbonate buffer solution of pH9.6；2) it is coated with micro reaction plate, every 100 μ L of hole, 4 DEG C overnight；3) 1 is washed with PBS buffer solution Time, every hole is added 1%BSA, 37 DEG C 1-2 hours；4) it is washed 1 time, is patted dry with PBS；5) primary antibody is added (after PBS 1:4000 dilution MAP antibody), be added coated reaction plate, every 100 μ L of hole are compared with PBS.37 DEG C of incubation 60min；6) it is added with 1% BSA diluted goat anti-rabbit igg-HRP, every hole 100 μ L, 37 DEG C of incubation 60min；7) color developing agent OPD, 37 DEG C of incubation 10-20 is added Minute；8) terminate liquid, every 50 μ l of hole is added；8) it reads at wavelength 492nm, is as a result indicated with OD mean value, as a result such as Fig. 4 institute Show, the binding force of MAP2 polypeptide and TNF-α holoprotein antibody is significantly stronger than MAP1.

(2) MAP antibody specificity is examined: by 1 × 10 after centrifuge washing⁹(the cell strain stability and high efficiency expression of H22 cell TNF-α is purchased from the Central China University of Science and Technology), the buffer suspension liquid vortex oscillator cell dispersion 5min of 1mL ice pre-cooling is added；In 4 DEG C with 3000r/min be centrifuged 5min；Supernatant is abandoned, 2 isometric × SDS is added as early as possible to extract total protein；It is made of total protein SDS-PAGE is compareed with commercialization TNF-α albumen；Western blot uses chemoluminescence method, and specific steps are referring to reagent Box specification carry out, antibody used be commercialization anti-tnf-alpha holoprotein antibody (be purchased from Cilag AG Ltd, ) or the anti-tnf-alpha B cell epitope MAP antibody that is purified Switzerland；Immunofluorescence observation gained band is complete with commercialization The kDa numerical value of band obtained by protein antibodies makees ratio, identifies the specificity of obtained purified antibody, and internal reference selects GAPDH.Quotient There is 1 gem-pure band at molecular weight about 17kD in product antibody；MAP1 and MAP2 goes out at molecular weight about 17kD Now clear band, compares through semi-quantitative analysis, and the gray value of MAP2 band is apparently higher than MAP1.According to commercial antibody explanation Book is TNF-α albumen at 17kD.And TNF-α recombinant protein antiserum and negative control occur in corresponding position without obvious band, See Fig. 5.

(3) MAP stimulated in vitro person monocytic cell (PBMC) proliferative capacity and IFN-β releasability are examined: using classics Standard 3H-TdR incorporation test.During the stimulation of lymphocyte receptor specific antigen is converted into lymphoblast, the synthesis of DNA It obviously increases, and the synthesis of its transforming degree and DNA are positively correlated, at this time by the precursor substance thymidine of synthetic DNA It is marked, is added in cultivating system with radioactive isotope (3H-TdR), that is, the lymphocyte being converted absorbs and mixes DNA points In son.After culture terminates, the exit dose of the 3H-TdR mixed in lymphocyte is measured to judge the transforming degree of lymphocyte. The result shows that MAP multiple antigenic peptide can significantly stimulate PBMC to be proliferated in vitro and discharge IFN-β, wherein MAP2 stimulates PBMC to discharge The ability of IFN-β is significantly stronger than MAP1.

Affinity test, the inspection of MAP antibody specificity, MAP vaccine pierce comprehensive analysis MAP antibody in vitro in vitro with MAP Swash the tests such as person monocytic cell's (PBMC) proliferative capacity and IFN-β releasability as a result, screening and identifying 54-66 bit amino Sour (VCFTLNKSQSNQE) is TNF-α B cell Dominant Epitopes.Therefore, multiple antigenic peptide MAP2 designs by MAP, uses molecule Measuring small and weak immunogenicity lysine is core matrix, is coupled together with sequence for the epitope polypeptide of VCFTLNKSQSNQE, The MAP for forming 8 branched peptide conformations is the multiple antigenic peptide vaccine of advantage B cell epitope.

Embodiment 2: the inside and outside immunology effect textual criticism of the multiple antigenic peptide of 1 the method for embodiment building.

Since rat and humanTNF-α have high homology, the present embodiment is textual criticism object with rat, according to embodiment The Dominant Epitopes that method described in 1 is screened, identified, it is then mostly anti-with the TNF-α B cell epitope MAP that its Dominant Epitopes synthesizes Former peptide vaccine carries out the inside and outside immunology effect textual criticism of rat, to prove more antigens of the 8 branched peptide conformations of the invention constructed Positive effect of the peptide in people and mouse ulcerative colitis.

Wherein, vitro effect textual criticism includes: influence of the MAP antibody to TNF-α Cytotoxicity in vitro l cell L929； The influence of the liver cancer cells HCC97H of MAP antibody molten broken 3H-TdR label external to TNF-α.Vivo effect textual criticism includes: TNF-α Influence of the B cell epitope MAP vaccine to model mice disease activity index；MAP vaccine is to model mice serum CA125, ESR, DAO Deng horizontal influence；The influence that MAP vaccine shifts model mice lymphonodi mesenterici；MAP vaccine is viscous to model mice colon Film pathological change, the expression of colonic tissue CAM 120/80 and the active influence of colonic tissue MPO.It is specific as follows:

1. BALB/c mouse is immunized in the subcutaneous multiple spot of MAP2,1 time every 2 weeks, totally 4 times, Freund ' s Freund's incomplete adjuvant and Th are logical It is used for booster immunization with auxiliary small peptide, collects animal blood serum with eye socket blood taking method in batches within immune latter 10 days every time, standard is indirect ELISA method measures and compares MAP antibody titer in each group (MAP2, TNF-α shell egg bletilla PBS control group) serum.First immunisation 2 Specific antibody can be detected after week, the average highest antibody titer that MAP2 immune antibody titer measures after 6 weeks is 1: 130000,8 tops Zhou Houda, Mean antibody titers 1:140000, TNF-α control group and PBS control group are without special Property antibody generate, see Fig. 6.The specific antibody that MAP2 is induced out is isolated and purified through Sephadase column chromatography collects IgG, and Zhongtai Bio-Chem. Co., Ltd., Hangzhou is entrusted to be separated with antibody affinity chromatography, specific polyclonal contained in purifying antiserum resists Body；Measuring gained antibody mean concentration with Coomassie brilliant blue after purification is 28.6mg/dL.

2. after the intervention of MAP antibody, observing TNF-α in vitro to the lethal effect of l cell L929 cell: giving birth to The good Mouse L929 cell of long status is adjusted to 10⁶The cell suspension of/mL, it is outstanding that 100 μ L cells are added in every hole in 96 well culture plates Liquid, 5%CO2,37 DEG C of overnight incubations；Abandon supernatant, every hole is added TNF-α and MAP antibody 5%CO2,37 DEG C of culture 16h, in abandoning Clearly, the concentration that 30 μ L are added in every hole is 500 μ g/mL violet staining 3-5min, carefully washes away crystal violet with flowing water, falls dry remnants Moisture, every hole are added 100 μ L of destainer, measure light absorption value at microplate reader 570nm.As a result as shown in fig. 7, as MAP antibody is dense The increase of degree, TNF-α gradually weaken (P < 0.05) to the lethal effect of l cell L929 cell.

3. after the intervention of MAP antibody, observation TNF-α is in vitro to the molten of human primary liver cancer's cell HCC97H of 3H-TdR label Broken rate: TNF-α receptor is widely distributed in kinds of tumor cells, the TNF-α receptor and TNF-α knot of certain tumour cell film surfaces After conjunction, the death of these tumour cells can lead to, it can be by detecting the killing ability to tumour cell, the life of Lai Fanying TNF-α Object activity.HCC97H liver cancer cells are first marked with 3H-TdR, and 3H-TdR is discharged to extracellular after being killed, and pass through measurement tumour The 3H-TdR of cell release shows that after the intervention of MAP antibody, TNF-α is substantially reduced the molten broken rate of HCC97H cell.

3. vivo immunization effect is investigated:

A, the foundation of mouse UC model: grope repeatedly with reference to the method for Kihara N etc., establish disease after being improved The more uniform DSS BALB/c mouse UC model of load, blank control group mouse often daily equal amount of distilled water stomach-filling conduct pair According to.

B, choose 18-20 grams of weight, 5-6 week old male BALB/c UC model mice 30, be divided into 3 groups, between group weight without Statistical difference.1st group 10, immunization method is that MAP vaccine is immunized in subcutaneous multiple spot, and immunizing dose is 50 μ g/, is exempted from for the first time Epidemic disease Freund's complete adjuvant, after booster immunization incomplete Freund's adjuvant three times, adjuvant and antigen 1: 1 is sufficiently mixed, and exempts from every other week Epidemic disease is primary, is immunized 4 times altogether；2nd group 10, with 50 μ g/ of Lysine substrate, only subcutaneous multiple spot is immune, and frequency and the 1st group of phase is immunized Together.Third group 10, immunization method is immune for subcutaneous multiple spot, and immunizing dose is 100 μ L physiological saline/only, and frequency and the 1st is immunized Group is identical.Start within 4 weeks after immune the mean disease activity index for recording each group mouse weekly, as the result is shown experimental mice disease Activity index is substantially less than matrix group or saline control group.Every group of experimental animal is 4 weeks after immune, each execution 5 in 8 weeks. Mouse fasting 12h, free water before collection of specimens.Ketalar and Su Mian Xin are made into anesthetic with l:l ratio, by 1mg/ Kg dosage intraperitoneal injection of anesthesia mouse to be put to death, dorsal position is fixed after anesthesia:

I, serum: eyeball of mouse is extractd with sterile tweezer, is taken a blood sample out of eye socket repeatedly with siphon pipe, blood about lml is taken, is packed into warp In the wet centrifuge tube of 1% heparin, it is stored at room temperature 30min, 4 DEG C of 3000r/min are centrifuged 10min, collect supernatant, dispense to 2 After sterile Eppendorf pipe, -80 DEG C of preservations, for detecting the content of serum D-LAC, ESR, CRP and DAO.As a result it prompts, After ulcerative colitis mouse inoculation MAP vaccine, D-LAC, ESR, CRP content and DAO content are significantly reduced in serum.

II, lymphonodi mesenterici Bacteria Culture observe bacterial translocation situation: opening abdomen rapidly after taking blood, take intestines under aseptic condition Mesentery lymph node about 200mg, is put into sterile grinder, and 2mL sterile saline is added, and hand lapping 2min takes 100ul to grind Grinding fluid is inoculated into blood agar culture dish.The results show that being vaccinated with the exedens mouse lymphonodi mesenterici Bacteria Culture of MAP vaccine Positive rate is significantly reduced compared with control group, and vaccine inoculation bacterial translocation situation is prompted to mitigate.

III, colonic tissue: exposure abdominal tissue, careful separation colon (from ileocecus to anus) are longitudinally cutd open along mesenterium Intestinal segment is opened, is rinsed with ice physiological saline, intestinal lumen contents is washed away, away from colonic tissue 2cm is taken at anus lcm, with 10% Fu Er Malin fixes, routine paraffin wax embedding, and continuous 4 μm of slices cut 3 altogether, the H&E dyeing and immunohistochemistry dye for histotomy Color.Be close to and take colonic tissue 10mg again, put the heat insulation tank that liquid nitrogen has been contained in merging after Eppendorf pipe into rapidly, after be put into -80 DEG C Refrigerator freezing saves, and the active measurement of colonic tissue MPO is used for, as a result as shown in figure 8, being vaccinated with the mouse (MP of MAP vaccine Group) colonic tissue MPO activity is substantially reduced compared with matrix group (M group) and TH control group.

(× 100, × 400) are observed, compare mouse colonic tissue under light microscopic after iv, colonic tissue sections observation: H&E dyeing Pathological change, as a result as shown in figure 9, the ulcerative colitis mouse Colon tissue karyocyte infiltration of inoculation MAP vaccine and gland It is substantially reduced that body destroys situation.

V, colonic tissue CAM 120/80 immunohistochemistry detects: using SP method, antibody used is rabbit-anti CAM 120/80 monoclonal antibody And adding the goat anti-rabbit igg working solution of biotin labeling, microscopically observation simultaneously takes the photograph piece, and cell membrane dyeing is sun in brown color person Property, the results are shown in Figure 10, the significant drop of ulcerative colitis mouse Colon tissue CAM 120/80 expression of inoculation MAP vaccine It is low.

By investigating data above it is found that the multiple antigenic peptide vaccine for the 8 branched peptide conformations that the present invention constructs is in prevention and treatment ulcer Property the colitis state of an illness exacerbation and repeatedly in have positive effect.

Detailed features and method of the invention that the present invention is explained by the above embodiments, but the invention is not limited to Above-mentioned detailed features and method do not mean that the present invention must rely on above-mentioned detailed features and method and could implement.Institute Belong to those skilled in the art it will be clearly understood that any improvement in the present invention, to material selected by the present invention and step etc. Effect replacement and increase, selection of concrete mode of auxiliary material and step etc. all fall within protection scope of the present invention and openly Within the scope of.

Claims (4)

1. a kind of B cell epitope of TNF-α albumen, which is characterized in that the amino acid sequence of the epitope is VCFTLNKSQSNQE。

2. a kind of prevention and treatment ulcerative colitis aggravates or multiple antigenic peptide repeatedly, which is characterized in that the multiple antigenic peptide is to pass through MAP design, is core matrix with lysine, is coupled together with sequence for the epitope polypeptide of VCFTLNKSQSNQE, forms 8 points The multiple antigenic peptide of branch peptide conformation.

3. epitope described in a kind of claim 1 preparation aggravate for preventing and treating ulcerative colitis or vaccine repeatedly in answer With.

4. a kind of multiple antigenic peptide as claimed in claim 2 preparation aggravate for preventing and treating ulcerative colitis or vaccine repeatedly in Application.