三种不同方法体外诱导CD4+T细胞向Th17细胞分化与增殖的效率

摘要

背景:研究认为Th17细胞参与了许多炎症性疾病的发生,但其体外诱导分化效率尚不明确.目的:比较采用3种不同方法在体外诱导CD4+T细胞向Th17细胞分化与增殖的效率.方法:分别采取以下3种方法体外诱导Th17细胞:①给予CD3及CD28抗体刺激.并在培养体系中加入白细胞介素6及转化生长因子β.培养3d.②在方法①的基础上加入白细胞介素1β及肿瘤坏死因子α,培养3d.③在方法②的基础上第3天洗去细胞因子,培养2d;再次给予CD3及CD28抗体刺激,并在培养体系中加入白细胞介素23,培养3d.结果与结论:3种方法培养后CD4+T细胞中Th17细胞的比例分别为(8.5±2.8)%,(26.9±4.3)%,(44.3±5.5)%,三组之间差异有显著性意义(P<0.01).提示在培养体系中加入白细胞介素1β、肿瘤坏死因子α及白细胞介素23,更有利于增加CD4+T细胞向Th17细胞分化的比例.%BACKGROUND: Th17 cells participate in the occurrence of many inflammatory diseases, but their induced differentiationefficiency remains poorly clear.OBJECTIVE: To compare the differentiation and proliferation efficiency from CD4+ calls to Th17 cells induced by three differentmethods.METHODS: Three methods to induce Th17 cells in vitro are as follows:(1) stimulation with CD3 and CD28 antibodies followed byaddition of interleukin (IL)-6 and transforming growth factor-p (TGF-3) in trie culture medium and finally culture for 3 days; (2)based on method (1), addition of extra IL-1β and tumor necrosis factor-a (TNF-a) and culture for 3 days; (3) based on method (2).Washing out previous cytokines on the 3rd day, culture for 2 days, stimulation with CD3 and CD28 antibodies again, addition ofIL-23 and finally culture for 3 days.RESULTS AND CONCLUSION: The proportion of Th17 cells in CD4'T cells was (8.5+2.8)%, (26.9±4.3)%, and (44.3±5.5)% afterinduced by three above-mentioned methods, respectively, with significant difference among these three methods (P < 0.01).These findings suggest that addition of IL-1β, TNF-α and IL-23 into the culture medium facilitates the induced differentiation fromCO4+ cells to Th17 cells.