Immunology Department, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.
In Vitro Cell Dev Biol Anim. 2011 Sep;47(8):581-92. doi: 10.1007/s11626-011-9444-1. Epub 2011 Aug 19.
Regarding discrepancies that exist among different studies which have tried to clarify critical factors in human Th17 cell differentiation, the aim of this study was to identify the best condition for human Th17 differentiation and to clarify the possible role of TGF-β in differentiation of these cells. Naïve CD4(+) T cells were isolated from cord blood samples and cultured either in X-VIVO 15 serum-free medium or RPMI 1640 containing 10% FBS. Purified cells were treated with different combinations of polarizing cytokines (TGF-β, IL-1β, IL-6, IL-23 and IL-21) followed by analysis of the expression of characteristic genes and their relevant cytokines by real-time quantitative RT-PCR and ELISA method, respectively. Our data indicate that a combination of TGF-β plus IL-6 and IL-23 cytokines in X-VIVO 15 serum-free medium could be applied as the best condition for developing human Th17 cells in compare with other studied cytokine treatments. It is shown that TGF-β could be considered as a positive regulator for human Th17 cell differentiation only if applied in average concentrations. Interestingly, polarizing treatments in absence of TGF-β, induced double-secreting Th17 cells which co-express IL-17 and IFN-γ whereas polarization in presence of TGF-β-induced single-secreting (only IL-17 expressing) Th17 cells.
关于不同研究在试图阐明人 Th17 细胞分化的关键因素方面存在差异，本研究旨在确定人 Th17 分化的最佳条件，并阐明 TGF-β 在这些细胞分化中的可能作用。从脐血样本中分离出幼稚 CD4(+) T 细胞，并在 X-VIVO 15 无血清培养基或含 10% FBS 的 RPMI 1640 中培养。用不同组合的极化细胞因子（TGF-β、IL-1β、IL-6、IL-23 和 IL-21）处理纯化细胞，然后通过实时定量 RT-PCR 和 ELISA 方法分别分析特征基因及其相关细胞因子的表达。我们的数据表明，在 X-VIVO 15 无血清培养基中，TGF-β 加 IL-6 和 IL-23 细胞因子的组合可作为开发人 Th17 细胞的最佳条件，与其他研究的细胞因子处理相比。结果表明，TGF-β 仅在平均浓度下可被视为人 Th17 细胞分化的正调节剂。有趣的是，在没有 TGF-β 的情况下进行极化处理会诱导双分泌 Th17 细胞，这些细胞共同表达 IL-17 和 IFN-γ，而在 TGF-β 存在的情况下进行极化会诱导单分泌（仅表达 IL-17）Th17 细胞。
