NOTCH signaling has been shown to play a role in the production of interleukin-22 (IL-22) by CD4+ T cells. Multiple T-helper (Th) cell populations secrete IL-22. Th22 (CD4+IL22+IFNγ−IL17A−) cells are a subgroup of CD4+ effector T cells that primarily generate IL-22. The regulatory mechanisms of the NOTCH signaling pathway involved in differentiation of the Th22 cell subset have not been completely elucidated. This study aimed to further explore the involvement of NOTCH signaling in Th22 differentiation. In vitro combination of IL-6, IL-23, and tumor necrosis factor-α (TNF-α) treatment with naïve CD4+ T cells established the Th22 cell induced model. NOTCH signaling was activated by jagged-1 and inhibited by (2S)-N-[(3,5-difluorophenyl) acetyl]-L-alanyl-2-phenyl]glycine 1,1-dimethylethyl ester (DAPT). HES-1 siRNA and HES-1 vector were employed to knock down and induce overexpression of HES-1 to investigate the effect of NOTCH signaling on the differentiation of CD4+T cells into Th22 cells. We observed that the proportion of Th22 cells, along with Hes-1, Ahr, and Il-22 mRNA and protein expression, was increased by both jagged-1 and overexpression of HES-1. On the other hand, after the combined cytokine treatment of cells, and exposure to jagged-1 and DAPT or HES-1 siRNA, there was a decrease in the Th22 cell proportion, mRNA and protein expression of HES-1, AHR, and IL-22. Our study demonstrates that HES-1 enhancement in AHR and IL-22 up-regulation of NOTCH signaling can promote the skewing of naïve CD4+T cells toward Th22 cells. Also, the results of our study show that HES-1 is a crucial factor in Th22 cell differentiation.

中文翻译：

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NOTCH-HES-1轴参与促进Th22细胞分化

NOTCH 信号已显示在 CD4+ T 细胞产生白细胞介素 22 (IL-22) 中发挥作用。多个 T 辅助 (Th) 细胞群会分泌 IL-22。Th22（CD4+IL22+IFNγ-IL17A-）细胞是主要产生IL-22的CD4+效应T细胞亚群。参与 Th22 细胞亚群分化的 NOTCH 信号通路的调控机制尚未完全阐明。本研究旨在进一步探索 NOTCH 信号在 Th22 分化中的参与。IL-6、IL-23 和肿瘤坏死因子-α (TNF-α) 与幼稚 CD4+ T 细胞的体外联合治疗建立了 Th22 细胞诱导模型。NOTCH 信号被 jagged-1 激活并被 (2S)-N-[(3,5-difluorophenyl) acetyl]-L-alanyl-2-phenyl] 甘氨酸 1,1-二甲基乙酯 (DAPT) 抑制。采用 HES-1 siRNA 和 HES-1 载体敲低并诱导 HES-1 过表达，以研究 NOTCH 信号对 CD4+T 细胞向 Th22 细胞分化的影响。我们观察到 Th22 细胞的比例以及 Hes-1、Ahr 和 Il-22 的 mRNA 和蛋白质表达都因 HES-1 的 jagged-1 和过表达而增加。另一方面，细胞在联合细胞因子处理后，暴露于 jagged-1 和 DAPT 或 HES-1 siRNA 后，Th22 细胞比例、HES-1、AHR 和 IL 的 mRNA 和蛋白表达降低-22。我们的研究表明，AHR 中的 HES-1 增强和 NOTCH 信号的 IL-22 上调可以促进幼稚 CD4+T 细胞向 Th22 细胞倾斜。此外，我们的研究结果表明，HES-1 是 Th22 细胞分化的关键因素。