PD-L2融合蛋白对角质形成细胞/T细胞共培养模型的作用

Effects of PD-L2-Ig in the co-culture model of keratinocytes and T lymphocytes

目的:探讨人PD-L2融合蛋白对体外模拟口腔扁平苔藓(OLP)局部发病机制的角质形成细胞/T细胞共培养模型的影响。方法:分离培养健康人的角质形成细胞、T细胞,建立共培养模型。实验组加入人PD-L2融合蛋白,阴性对照组加入人IgG1,空白对照组加入含0.1%BSA的PBS,分别继续共培养72h。流式细胞术检测共培养模型中T细胞凋亡及生长周期的改变,MTT法检测T细胞增殖状态,ELISA检测共培养上清中IL-2、IFN-γ表达水平。结果:人PD-L2融合蛋白抑制了共培养模型中T细胞增殖及细胞因子IL-2、IFN-γ的表达,促进了T细胞凋亡,但是对T细胞生长周期没有明显的影响。结论:人PD-L2融合蛋白对体外角质形成细胞/T细胞共培养模型中T细胞增殖活化有一定负性作用,可能是该融合蛋白拮抗了PD-L2与其第二受体配接后促进T细胞增殖的效应,人PD-L2融合蛋白有望成为OLP新的有效的免疫治疗方法。

Objective：To investigate the effects of human PD-L2-Ig on the T lymphocytes in the co-culture model of keratinocytes and T lymphocytes which mimics local ccondition of oral lichen planus in vivo.Method：Keratinocytes and T lymphocytes from healthy volunteers were obtained respectively and co-cultured 72 h in vitro since being stimulated by INF-？.Three groups of the above-mentioned model were cultured 72 h followed by adding human PD-L2-Ig,human IgG1 and PBS separately.FCM was employed to detect the T lymphocytes＇ apoptosis rate and the change of cell-cycle;the T lymphocytes＇ proliferation was determined by MTT method;and ELISA was used to test the expression of IL-2、IFN-γ.Result：Compared with the control groups,PD-L2-Ig inhibited the T lymphocytes＇ proliferation and the expression of IL-2 and IFN-γ,and accelerated the cells＇ apoptosis rate,but didn＇t influence the cell-cycle.Conclusion：PD-L2-Ig could regulate the T lymphocytes＇ functions negatively resulted from the PD-L2-Ig,as an antagonist,maybe neutralized the positive effect of PD-L2 on the T lymphocytes,so the PD-L2-Ig will be an new interesting approach for the treatment of OLP.