Studies On The Effect Of CKZ In Transplantation Immunology And Its Mechanisms

Chapter 1 Relativity between prolonged survival of skin allograft by ckz and the CD4~+CD25~+ regulatory T cellsAim: To study the relativity between prolonged survival of skin allograft by ckz and CD4~+CD25~+ regulatory T cells.Methods: Skin allograft and isograft model in mouse were established ; three color fluorescent staining together with flow cytometry were used to analyze the change of CD4~+CD25~+regulatory T cells。Results: These CD4~+CD25~+regulatory T cells in allograft plus CKZ were significantly higher than the control (p<0.01).Conclusion:CKZ could prolong the allograft survival, suggesting that the enhancement of CD4~+CD25~+regulatory T cells maybe associated with the immunosuppression effect of CKZ for allograft.Chapter 2 Effect of CKZ on GVHD in mouse allogeneic bone marrow transplantationAim: To study the effect of CKZ on graft-versus-host disease(GVHD) in mouse allogeneic bone marrow transplantation.Methods: Lethally irradiated BALB/c(H-2d) mice were transplanted with C57BL/6(H-2b) bone marrow containing donor peripheral T cells, i.p CKZ and/or CSA. GVHD and survival rates were then observed.Results: In the group of transplanted with CKZ infusion, aGVHD were induced only in 60% of 0.25mL ckz group and 70% of 0.5mL CKZ group; then aGVHD were induced only in 30% of CSA group; the incidence of GVHD was evidently lessened compare with the bone marrow add T cells without CKZ and CSA infusion groups (P<0.05), and survival rates was significantly higher than that bone marrow add T cells without CKZ and CSA infusion P<0.0\. these CD4+CD25+regulatory T cells in allo-BMT plus CKZ were significantly higher than the control (p<0.05).Conclusion: In mouse allogeneic bone marrow transplantation, CKZ could partly reduce the incidence of GVHD, and increase survival rates, suggesting that the enhancement of CD4+CD25+regulatory T cells associated with the immunosuppression effect of CKZ for allo-BMT.and the pattern of cytokine expression(IL-10 went up) is central to the mechanisms which regulate the development of immune tolerance to allo-BMT.Chapter 3 Effects of CKZ on mouse immune response in vitroAim: To study the effects of CKZ on mouse immune response in vitro.Methods: in vitro, ConA plurs CKZ were used to activate lymphocytes,then expression of CD69 and CD25 on T cells were detected and analyzed by flow cytometry.Results: CKZ had a inhibitory effect on the expression of CD69 of activitied T cells,but had a stimulatory effect on the expression of CD25 of activitied T cells.Conclusion :CKZ could inhibited strong immune response on mouse,and promote establishment maintenance of immune tolerance.Chapter 4 Experimental study on the acceleration of the hemato -immunological reconstitution of the nonlethally irradiated mouse by CKZAim: To study the effect of the CKZ on the hematological and immunological reconstitution of mice.Methods: The nonlethally irradiated model in mouse were established ; auto-cytometry and flow cytometry were used to analyze the change of the hemato-immunological reconstitution.Results: The hemato-immuno logical reconstitution of the nonlethally irradiated mice by CKZ were significantly higher than the control of rhg-csf; but the bone marrow stem cell reconstitution of the nonlethally irradiated mice by CKZ were lower than the control of rhg-csf.Then the CKZ group survival time significantly longer than the control.Conclusion: CKZ could acceleration of the hemato-immuno logical reconstitution of the nonlethally irradiated mice.