【Abstract】Objective To explore the feasibility that the recipient against donor antigen-specific T lymphocytes clones are formed,and the suicide genes are induced into the clone. In the end it may induce the transplanted-organ tolerance. Methods The recipient rats were immunized by the donor rats-splenocytes, then the recipient’s T cell were isolated, purified and diluted with limited-dilute methods into single cell.The T cells were cultured by adding raise cells,ConA or IL2 under the different concentration and at last the T cell clone were formed. Results A stable recipient against donor antigen-specific T cell clone was established. The difference among the different immune groups was very significant(tgt;t0.05). The T cell clone was not formed without raise cells or only with raise cells. The rare clone could be formed by ConA stimulation but without IL-2. The clone-forming rate was associated with concentration of IL-2. ConclusionThe mature T cell is able to proliferate to form clone when the condition is suitable. The recipient is immunized with donor rats spleen cellular antigen. The T cells clones selected in the end are donor antigen-specific.
To diminish the specific lymphocytes that responsive to the rejection of allograft. Anti-rat CD4,CD8 monoclonal antibodies and trichosnthin (TCS) was conjugated to immunotoxin by heterobifunctional reagent SPDP, 2-IT. The free TCS was removed from conjugates mixture by a column of Sephacryl S-200. The SDS-PAGE and cytotoxic assay was used to measure the biological activity of immunotoxin. SDS-PAGE showed the immunotoxin, free McAb and TCS were in the mixture of conjugation, and the free TCS can be separated by Sephacryl S-200. In Vitro, the lymphocytes of rat can be killed by antiCD4,antiCD8 immunotoxin. The kill capability was relay to the amount of immunotoxin. The authors consider that the immunology unresponsiveness can be induced by antiCD4,antiCD8 immunotoxin. That was useful in induced transplantation tolerance.