Production, Identification, Animal Experiment Of Adjuvant Vaccine Of Anti-idiotypic Antibody Of Anti-human Bladder Carcinoma And Its Preliminary Clinical Application

Based on Jerne's immune network study,Nisonoff and Lamoyi pointed out that a kind of anti-idiotypic antibody(Ab2) which acted as epitope of antigen could connect with antibody(Ab).In fact,it could take the place of antigen to induce specific response.This reminded us that anti- idiotypic antibody could be used for active immune therapy.Traditional therapeutic method.such as chemotherapy and radiotherapy couldn't kill all the tumor cells in patients.This was why tumor could recur after clinical cure.Tumor treatment should modulate human immune condition,changed the living surroundings of tumor and inhibited tumor grow or reversed its growth. So active immune therapy was very important.Anti-idiotypic antibody could simulate antigen and regulate immunity. At the same time.it could overcome human immune inhibition or break up immune tolerance.lt had a giant potential in tumor therapy.In recent years,patients of melanocytoma and rectal carcinoma had been treated with active immune therapy in foreign countries and many other anti-idiotypic antibody had been produced.But there was no anti-idiotypic antibody of bladder carcinoma.Bladder carcinoma antigen (Ag)was drawn from human bladder transitional cell carcinoma by use of even grind and gel.Monoclonal antibody of anti-human bladder carcinoma was produced by use of bladder carcinoma antigen through B-lymphocyte hybridoma technology. F2 fragment from monoclonal antibody of anti-human bladder carcinoma was prepared through pepsin cleavage.Anti-idiotypic antibody of anti-human bladder carcinoma(Ab2) was produced by the Fb )2 fragment from monoclonal antibody of anti-human bladder carcinoma immuning rabbits.Its serum was purified through gel.Theresults of the experiments showed that: monoclonal antibody of anti-human bladder carcinoma was Igd subclass through agar diffusion test.Indirect ELISA showed that:the reaction between monoclonal antibody of anti-human bladder carcinoma and bladder transitional cell carcinoma was positive,while the reaction between monoclonal antibody of anti-human bladder carcinoma and bladder adenosarcoma.bladder squamous cell carcinoma,normal bladder tissues was negative.The hybridoma of monoclonal antibody of anti-human bladder carcinoma was treated with routine method of preparation of chromosome.The number of chromosome of a single hybridoma cell was 96-106.All the above-mentioned results showed that monoclonal antibody of anti-human bladder carcinoma was the antibody aimed at bladder transitional cell carcinoma.The reaction of anti-idiotypic antibody of anti-human bladder carcinoma was positive with monoclonal antibody of anti-human bladder carcinoma,while was negative with murine gamma globulin through indirect ELISA test,Neutralization test showed that the F(ab)2 fragment of monoclonal antibody of anti-human bladder carcinoma could neutralize anti-idiotypic antibody of anti-human bladder carcinoma. So we thought anti-idiotypic antibody of anti-human bladder carcinoma was aimed at the idiotype of variable region of monoclonal antibody of anti-human bladder carcinoma.From competitive inhibition assays,a series of dilution bladder carcinoma antigen was added to the well of the plate coated with monoclonal antibody of anti-human bladder carcinoma,after incubation,anti-idiotypic antibody of anti-human bladder carcinoma and rabbit IgG was added respectively to the plate.The result demonstrated that the OD number in adding anti-idiotypic antibody of anti-human bladder carcinoma increased accompanying the dose of bladder carcinoma antigen decreased,while the OD number of rabbit IgG was low withouttheregulation.lt indicated that anti-idiotyic antibody of anti-human bladder carcinoma competitvely bind monoclonal antibody of anti-human bladder carcinoma with bladder carcinoma antigen. A series of dilution anti-idiotypic antibody of anti- human bladder carcinoma and rabbit IgG was mixed respectivey with monoclonal antibody of anti-human bladder carcinoma. After incubation, they were added to the plate coated with bladder carcinoma antigen.The result s...