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目的:探讨雷帕霉素(rapamycin,RPM)对人外周血T淋巴细胞上B、T淋巴细胞弱化子(BTLA)表达的影响,为其在器官移植和自身免疫性疾病中的应用提供实验依据。方法:采用Ficoll密度梯度离心法分离人外周血单核细胞(PBMCs),然后利用免疫磁珠分离技术分离PBMCs中的T细胞。用刀豆蛋白A刺激、活化人外周血T细胞。用MTT比色法测定T细胞的增殖。用ELISA法测定细胞培养上清液中IL-2、IFN-γ的水平。用流式细胞术检测T细胞上BTLA的表达。结果:用不同浓度(10~1 000 mL)的RPM处理的T淋巴细胞上BTLA的表达无显著差别;而与未经RPM处理组比较差别显著(P<0.01)。ELISA检测结果显示不同浓度的RPM与未处理组相比均能显著抑制IL-2和IFN-γ的分泌(P<0.01),并可以剂量依赖的方式显著抑制T淋巴细胞增殖(P<0.01)。结论:RPM对BTLA的表达影响较小,而对T淋巴细胞增殖和IL-2、IFN-γ的分泌有较强的抑制作用,提示RPM适宜用于器官移植排斥反应和自身免疫性疾病的治疗。
Objective: To investigate the effect of rapamycin (RPM) on the expression of B and T lymphocyte attenuator (BTLA) on human peripheral blood T lymphocytes and to provide an experimental basis for its application in organ transplantation and autoimmune diseases . Methods: Peripheral blood mononuclear cells (PBMCs) were isolated by Ficoll density gradient centrifugation. T cells in PBMCs were isolated by immunomagnetic beads. Stimulated with concanavalin A to activate human peripheral blood T cells. T cell proliferation was measured by MTT colorimetry. The levels of IL-2 and IFN-γ in the cell culture supernatants were determined by ELISA. The expression of BTLA on T cells was detected by flow cytometry. Results: There was no significant difference in the expression of BTLA between T lymphocytes treated with different concentrations of RPM (10-1 000 mL), but not significantly different with those without RPM (P <0.01). The results of ELISA showed that RPM of different concentrations significantly inhibited the secretion of IL-2 and IFN-γ (P <0.01) and inhibited the proliferation of T lymphocytes in a dose-dependent manner (P <0.01) . CONCLUSION: RPM has little effect on the expression of BTLA, but has a strong inhibitory effect on the proliferation of T lymphocytes and the secretion of IL-2 and IFN-γ, suggesting that RPM is suitable for the treatment of organ transplant rejection and autoimmune diseases .