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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 effect of human PD-L2 fusion protein on keratinocyte / T cell co-culture model in vitro to simulate the local pathogenesis of oral lichen planus (OLP). Methods: Human keratinocytes and T cells were isolated and cultured to establish co-culture model. Human PD-L2 fusion protein was added to the experimental group, human IgG1 was added into the negative control group, PBS containing 0.1% BSA was added to the blank control group, and co-cultured for 72 hours respectively. Flow cytometry was used to detect the changes of T cell apoptosis and growth cycle in the co-culture model. The proliferation status of T cells was detected by MTT assay. The levels of IL-2 and IFN-γ in the co-culture supernatants were detected by ELISA. Results: Human PD-L2 fusion protein inhibited the proliferation of T cells and the expression of cytokines IL-2 and IFN-γ in the co-culture model and promoted the apoptosis of T cells. However, they did not affect the T cell growth cycle significantly. CONCLUSION: Human PD-L2 fusion protein has a negative effect on T cell proliferation and activation in a keratinocyte / T cell co-culture model in vitro. It may be that the fusion protein antagonizes the binding of PD-L2 and its second receptor to promote T The effect of cell proliferation, human PD-L2 fusion protein is expected to become the new effective immunotherapy for OLP.