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目的研究γ干扰素(IFN-γ)处理对C57BL/6小鼠树突状细胞(DC)生物学特性及功能的影响。方法在DC培养过程的早期(第2天)或晚期(第5天)加入20 ng/m L IFN-γ进行处理,第7天加入脂多糖(LPS)刺激DC成熟,流式细胞术检测细胞表面分子CD11c、CD80、CD86的表达。为检测DC功能,将其与BALB/c小鼠来源的淋巴细胞进行共培养,5,6-羧基荧光黄二乙酸盐N-琥珀酰亚胺酯(CFSE)标记法检测其刺激异基因淋巴细胞增殖的能力、流式细胞术检测其诱导调节性T细胞(Treg)产生的能力。结果与对照组相比,早期IFN-γ处理组的DC数量减少、分化受到抑制;虽然表面共刺激分子的表达无明显变化,但是却明显拮抗LPS刺激其成熟;刺激异基因淋巴细胞增殖的能力明显减弱;诱导产生更多的Treg。与对照组相比,晚期IFN-γ处理的DC数量及分化无变化;表面共刺激分子CD86及CD80表达升高;LPS刺激DC成熟及同种异体混合淋巴细胞反应的结果相似,但是诱导Treg产生的能力增加。结论早期IFN-γ处理组和晚期IFN-γ处理组的DC生物学特性及功能明显不同。
Objective To investigate the effects of IFN-γ on the biological characteristics and function of dendritic cells (DCs) in C57BL / 6 mice. Methods The cells were treated with 20 ng / mL IFN-γ in the early (day 2) or late (day 5) DC culture. DC maturation was induced by adding lipopolysaccharide (LPS) on the 7th day and the cells were detected by flow cytometry Expression of surface molecules CD11c, CD80, CD86. To detect DC function, BALB / c mouse-derived lymphocytes were co-cultured with 5,6-carboxyfluorescein diacetate N-succinimidyl ester (CFSE) to detect the proliferation of allogeneic lymphocytes The ability of induced Tregs (Tregs) was examined by flow cytometry. Results Compared with the control group, the number of DCs in the early IFN-γ treatment group was decreased and the differentiation was inhibited. Although the expression of the costimulatory molecules did not change significantly, they significantly antagonized the LPS-stimulated maturation and stimulated allogeneic lymphocyte proliferation Significantly weakened; induced more Tregs. Compared with the control group, there was no change in the number and differentiation of DCs treated with advanced IFN-γ; the expressions of CD86 and CD80 on the surface were increased; the results of LPS-stimulated DC maturation and allogenic mixed lymphocyte reaction were similar but Treg production was induced The ability to increase. Conclusion The biological characteristics and functions of DC in early IFN-γ group and late IFN-γ group are obviously different.