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目的:建立相关角质形成细胞/T细胞共培养模型,以模拟口腔扁平苔藓(oral lichen planus,OLP)局部损害的免疫应答环境,为进一步体外研究OLP提供较可靠的细胞模型。方法:采用酶消化法培养正常口腔黏膜角质形成细胞,利用磁珠阴性分选法获取正常人外周血T淋巴细胞,共培养后,利用MTT法测定T细胞增殖,利用ELISA法测定细胞因子IFN-γ的表达。采用SAS.6.12软件包中的成组设计资料的t检验进行统计学分析。结果:表达PD-L1、PD-L2的角质形成细胞/T细胞共培养组T细胞的增殖程度比T细胞单独培养组高(P<0.05),共培养组较T细胞单独培养组IFN-γ分泌水平升高(P<0.01)。结论:表达PD-L1、PD-L2的角质形成细胞/T细胞共培养模型能在一定程度上模拟OLP局部损害的免疫应答环境。
OBJECTIVE: To establish a related keratinocyte / T cell co-culture model to simulate the local immune response environment of oral lichen planus (OLP), and to provide a more reliable cell model for the further study of OLP in vitro. Methods: Normal oral mucosal keratinocytes were cultured by enzymatic digestion, T lymphocytes were obtained from normal human peripheral blood by magnetic beads negative sorting. After co-cultured, the proliferation of T cells was measured by MTT assay. The levels of IFN- γ expression. Statistical analysis was conducted using the t-test of the group design data in the SAS.6.12 package. Results: The proliferation of keratinocyte / T cell co-culture group expressing PD-L1 and PD-L2 was higher than that of T-cell alone group (P <0.05). Compared with T-cell alone group, IFN- γ Secretion increased (P <0.01). CONCLUSION: The keratinocyte / T cell co-culture model expressing PD-L1 and PD-L2 can simulate the immune response environment of local lesion of OLP to some extent.