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

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3 d。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3 d。③在方法②的基础上第3天洗去细胞因子，培养2 d；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3 d。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。

Comparison of differentiation and proliferation efficiency from CD4+ cells to Th17 cells induced by three different methods

BACKGROUND: Th17 cells participate in the occurrence of many inflammatory diseases, but their induced differentiation efficiency remains poorly clear. OBJECTIVE:To compare the differentiation and proliferation efficiency from CD4⁺ cells to Th17 cells induced by three different methods.METHODS:Three methods to induce Th17 cells in vitro are as follows: (1) stimulation with CD3 and CD28 antibodies followed by addition of interleukin (IL)-6 and transforming growth factor-β (TGF-β) in the culture medium and finally culture for 3 days; (2) based on method (1), addition of extra IL-1β and tumor necrosis factor-α (TNF-α) 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 of IL-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)% after induced 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 from CD4⁺ cells to Th17 cells.