| Objective: Through detecting the secretion of IFN-γin Thl and IL-4 in Th2 which included healthy volunteers, cyclosporine A (CsA) treated and tacrolimus (FK506) treated patients after renal transplantation for three years, we intended to investigate the effects of CsA and FK506 on the differentiations of Thl and Th2. At the same time, by observing abve-mentioned parameters in CsA-treated and FK506-treated allo-kidney recipients, the thesis compared the different immunosuppressive effects between the two calcineurin inhibitors. This would help us to correctly evaluate the distinction on anti-rejection of CsA and FK506, so that rejection could be precluded and more effective immunosuppressive protocol could be formulated after kidney transplantation.Methods: Peripheral blood (Heparin for anticoagulation) was stimulated with phorbol myristate acetate(PMA) and ionomycin for 4 hours, in the presence of monensin. CD3 and CD8 monoclonal antibodies were added to conjugate the correspondent antigens on T-cells surface. IFN-γand IL-4 antibodies were conjugated the correspondent antigens in T-cells after hemolysis and permeation. Using CD8 and IFN-γor CD8 and IL-4 to build histograms in the gate of CD3+ lymphocytes, and finding CD3+/CD8-/IFN-γ+ cells (Th1) and CD3+/CD8-/IL-4+ cells (Th2) by flow cytometry There were three groups: control group, CsA group, FK506 group. Results were used to make statistical description and analysis with SPSS 12.0. Mean±standard deviation were employed to express the differentiation of Thl and Th2. The diversity of Th1 and Th2 differentiation under contribution of CsA and FK506 were analyzed with T tests and Nonparametric tests .Results: 1.The percentage of Th1 cells on patients was lower than health-control group, and there were differences on them(P﹤0.05), at the same time, the percentage of Th1 cells on pre-FK506 group was much lower than health-control group, and also there were significant differences on them(P﹤0.05), but the percentage of Th1 cells didn't change clearly in pre-CsA group(P﹥0.05), even Th1 cells on pre-FK506 group were obviously lower than that on pre-CsA group(P﹤0.05).There were no distinct differences among health-control group, all patients group, pre-CsA and pre-FK506 in Th2 cells(P﹥0.05). 2. The expression of Th1 did not change markedly beween post-CsA and post-FK506, nor did in Th2 cells of post-CsA and post-FK506(P﹥0.05).3 . The differentiation of Th1 in post-CsA was much lower than that in pre-CsA, as the same result of post-FK506 and pre-FK506(P﹤0.05), but there were no significant differences in the differentiation of Th2 cells between pre-CsA and post-CsA or between pre-FK506 and post- FK506(P﹥0.05). 4.The difference of pre-CsA and post-CsA in Th1 was visibly higher than that on pre-FK506 and post-FK506(P﹤0.05).Conclusions: 1. The differentiation of Th2 cells are not affected by the two immunosuppressive agents. 2. Tacrolimus and cyclosporine A distinctly inhibit the differentiation of renal allograft patients Th1 at the highest peak of the concentraiton, and the effects of two immunosuppressive agents are identical. 3. When comparing the frequencies of Th1 cells in FK506 and CsA-treated patients, we find that IFN-γexpressing T cells are restrained even prior to the morning intake of tacrolimus, but not for cycloporine A(CsA). |
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