1Experimental Research Of Teh Immune Tolerance Induction By Mouce Splenocytes 2Living-related Partial Orthotopic Liver Transplantation:3cases Review

Although the application of transplantation has saved the lives of numerous patients in tenninal stages of diseases, the serious rejection after transplantation is still the main cause that affects survival time of recipient. Although powerful immunosuppressant drugs such as CsA and FK506 can suppress rejection, their side effects such as hyperpiesia, hepatorenal toxicity, severe infection, malignant degeneration limit their long time usage. Nowadays the key method to avoid or alleviate these side effects is induction of immune tolerance or specific low immunoreaction state with simultaneous maintenance of immune response against other antigens (including anticancer, anti-infection and so on). According to mechanism of immune tolerance, we designed the experiment to induce anti-idiotypic antibody, which has the same antigenicity as the graft antigen in recipient prior to transplantation, and observed effect on the HAGR. Aims: By the induction of anti-idiotypic antibody that has the same antigenicity as the graft antigen in recipient prior to transplantation, we 7 probe to investigate its tolerance induction effect on transplantation in mice. Methods: 1. Isotypic heterostrain antibody (AbI) Induction: BalbIc mice were immunized with C57BL16 splenocytes to prepare homogeneous hetero strain antibody (Ab 1). 2. Anti-idiotypic antibody (Ab2) Induction: Immunoglobulin was extracted grossly from Ab 1 serum by saturated ammonium sulfate salting out. Then immunoglobulin was purified by DEAE-Sephadex A-50 column. Abl was cross-linked to KLII, and Abl-KLH plus Freund抯 adjuvant were used to induce anti-idiotypic antibody (Ab2) in Balb/c mice. Ab2 was detected by Sandwich ELISA. 3. The immune tolerance induction effect in mice by Ab2: Skin graft was performed in mice, the survival time of transplanted graft niicrolymphocytotoxicity and mixture lymphocytes culture were investigated to observe the effect of Ab2 induction on mice immunoreaction. Results: 1. Determination of the titer of Abl and Ab2: By using component-dependent cytotoxicity test, the titer of antiserum is> 1: 400. By using sandwich ELISA, the A4~ of the Ab2 induction mice is over two times than that of the contol group. 2. Influence of Ab2 induction on skin graft: The survival time of skin 8 graft of Ab2 induction group is longer than control group significantly (P <0.01). 3. Influence of Ab2 serum on microlymphocytotoxicity: the positive cell rate of the group containing Ab2 serum was lower than control group, the difference was significant (P < 0.01). 4. Influence of Ab2 induction on mixture lymphocytes culture: The A570,~in Ab2 induction group is lower than control group is 0.35 ?.04, the difference was significant (P < 0.01). Conclusions: 1. Isotypic heterostrain antibody (Abl) can be successfully produced by immuning Balb/c mice with C57BL/6 mice splenocytes. Abl=KLH plus Freund抯 adjuvant can induce anti-idiotypic antibody (Ab2) in Balb/c mice successfi.tlly. 2. Compared with those in control group, Ab2 induced in donor can prolong the survival time of graft, and suppress other reje...