不同刺激剂对淋巴细胞中PD-1与TIM3蛋白表达的影响

Effects of different stimulators on PD-1 and TIM3 expression in lymphocytes

目的研究不同刺激剂对人外周血淋巴细胞中PD-1与TIM3蛋白表达的影响。方法采用密度离心法分离人外周血淋巴细胞,并通过Western blot检测在抗CD3/CD28抗体、PHA、SEB、Con A4种不同刺激剂下人外周血淋巴细胞PD-1与TIM3总蛋白的变化情况,并通过流式细胞技术检测4种刺激条件下T淋巴细胞及其CD3^+CD4^+、CD3^+CD8^+亚群膜表面的PD-1与TIM3的改变。结果 Western blot结果显示,4种刺激剂均能刺激总的PD-1蛋白的表达,减少总TIM3蛋白的表达。流式细胞分析仪结果显示,在4种刺激剂作用下,人外周血T淋巴细胞表面及其CD3^+CD4^+、CD3^+CD8^+T细胞亚群膜表面的PD-1与TIM3蛋白表达均呈现升高趋势。结论 4种常见刺激剂均可诱导总PD-1蛋白和细胞膜上PD-1蛋白的表达,降低TIM3总蛋白的表达以及诱导淋巴细胞表面TIM3蛋白的表达。其中PHA较其他3种刺激剂展现出较好的剂量关系。

Objective To study the effects of different stimulators on PD-1 and TIM3 expression in human peripheral blood lymphocytes. Methods Human peripheral blood lymphocytes were isolated by density centrifugation. Four different stimulators used are anti-CD3/CD28 antibody, PHA, SEB and ConA, respectively. Total PD-1 and TIM3 expression in human peripheral blood lymphocytes under four different conditions were evaluated by Western blot, while PD-1 and TIM3 expression on the membrane were detected by flow cytometry. Results Western blot results indicated that total PD-1 expression was increased, while total TIM3 expression was decreased upon treatment with all the four stimulators in T lymphocytes. However, Flow cytometry results indicated that PD-1 and TIM3 expression on the T lymphocytes membrane were increased under these four conditions. We also noticed that PD-1 and TIM3 expression were up-regulated in T cell subpopulation CD3^+CD4^+, CD3^+CD8^+ cells. Conclusion All the four common stimulators could induce PD-1 and TIM3 expression on the membrane, while total expression of the proteins shows different expression pattern. Compared with other stimulators, PHA induces better dose-dependent response in human peripheral blood lymphocytes.