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目的研究含噻唑烷-4-酮的糖类衍生小分子免疫刺激剂CH2a对人iNKT细胞功能的影响。方法分离健康人外周血单个核细胞(PBMC),经α-半乳糖神经酰胺(α-Galcer)和IL-2体外扩增后,利用磁珠分选分离纯化得到恒定型自然杀伤T细胞(iNKT细胞)。5(6)-羧基二乙酸荧光黄琥珀酰亚胺酯(CFDA-SE)标记的纯化iNKT细胞,与CH2a共孵育后,流式细胞术检测细胞增殖和分代,IFN-γ和IL-4的表达;MTT法定量检测细胞增殖率及对K562细胞的杀伤活性。ELISA检测CH2a处理细胞培养基中IFN-γ和IL-4水平。结果 CH2a能显著促进IL-2引起的iNKT细胞体外增殖;提高IFN-γ和IL-4的生成量及IFN-γ/IL-4的比值;并显著增强iNKT细胞的杀伤活性。结论 CH2a有可能通过诱导iNKT分泌Th1类的细胞因子,调节T细胞向Th1分化,从而增强细胞免疫功能;同时显著提高iNKT的细胞毒功能。
Objective To investigate the effect of thiazolidin-4-one-containing carbohydrate-derived small molecule immunostimulatory agent CH2a on the function of human iNKT cells. Methods Peripheral blood mononuclear cells (PBMCs) were isolated from human peripheral blood mononuclear cells (PBMCs). After expanded in vitro by α-Gal-Ce and IL-2, the natural killer T cells cell). The purified iNKT cells labeled with CFDA-SE and 5 (6) -carboxyl diacetate were co-incubated with CH2a. The proliferation and differentiation of IFN-γ and IL-4 were detected by flow cytometry MTT assay was used to detect cell proliferation rate and K562 cell killing activity. The levels of IFN-γ and IL-4 in CH2a-treated cell culture medium were detected by ELISA. Results CH2a could significantly promote the proliferation of iNKT cells induced by IL-2, increase the production of IFN-γ and IL-4 and the ratio of IFN-γ / IL-4, and significantly enhance the killing activity of iNKT cells. Conclusions CH2a may induce Th1 cytokines secretion by regulating iNKT secretion and regulate the differentiation of T cells into Th1 cells, thereby enhancing cellular immune function and significantly increasing the cytotoxic function of iNKT.