Liu Yanling, Wang Hongmei, Yu Yanrong, Yuan Keng, Zhang Yujuan, Min Weiping
Institute of Immunotherapy, Nanchang University, Nanchang 330006; Jiangxi Academy of Medical Sciences, Nanchang 330006, China.
Institute of Immunotherapy, Nanchang University, Nanchang 330006, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2016 Aug;32(8):1009-13.
Objective To investigate the effect of IFN-γ treatment on the biological characteristics and functions of C57BL/6 murine dendritic cells (DCs). Methods In the process of DC culture, 20 ng/mL IFN-γ was added in the DCs at the early (day 2) or late (day 5) stage, and on day 7, LPS was added to stimulate DC maturation. The expressions of DC surface molecules CD11c, CD80 and CD86 were determined by flow cytometry. To analyze cell functions, DCs were co-cultured with BALB/c mouse-derived lymphocyte cells. The 5, 6-carboxyfluorescein diacetate N-succinimidyl ester (CFSE) labelling was used to detect their ability to stimulate allogeneic lymphocyte proliferation and flow cytometry was used to measure their ability to induce the production of regulatory T cells (Tregs). Results Compared with the control group, the early IFN-γ treatment group had decreased DC number and inhibited cell differentiation; though there was no difference in the expression of co-stimulatory molecules, early IFN-γ treatment resisted the stimulatory effect of LPS on DC maturation, weakened the ability to stimulate allogeneic lymphocyte proliferation and enhanced the ability to induce more Tregs. Compared with the control group, the late IFN-γ treatment group showed no change in DC number and differentiation; the expressions of co-stimulatory molecules CD86 and CD80 were upregulated; the results of DC maturation and mixed allogeneic lymphocyte reaction stimulated by LPS were similar to those in the control group, but its ability to induce Tregs was stronger. Conclusion DCs treated with IFN-γ at early stage and those at late stage showed obvious difference in biological characteristics and functions.
目的 探讨干扰素-γ（IFN-γ）处理对C57BL/6小鼠树突状细胞（DCs）生物学特性及功能的影响。方法 在DC培养过程中，于早期（第2天）或晚期（第5天）向DCs中加入20 ng/mL IFN-γ，第7天加入脂多糖（LPS）刺激DC成熟。采用流式细胞术检测DC表面分子CD11c、CD80和CD86的表达。为分析细胞功能，将DCs与BALB/c小鼠来源的淋巴细胞共培养。采用5,6-羧基荧光素二乙酸琥珀酰亚胺酯（CFSE）标记检测其刺激同种异体淋巴细胞增殖的能力，并用流式细胞术检测其诱导调节性T细胞（Tregs）产生的能力。结果 与对照组相比，早期IFN-γ处理组DC数量减少，细胞分化受抑制；共刺激分子表达虽无差异，但早期IFN-γ处理可抵抗LPS对DC成熟的刺激作用，减弱刺激同种异体淋巴细胞增殖的能力，并增强诱导更多Tregs的能力。与对照组相比，晚期IFN-γ处理组DC数量及分化无变化；共刺激分子CD86和CD80表达上调；LPS刺激的DC成熟及混合同种异体淋巴细胞反应结果与对照组相似，但其诱导Tregs的能力更强。结论 早期和晚期用IFN-γ处理的DCs在生物学特性及功能上存在明显差异。
