Qin Ling, Qiu Ke-Zi, Hu Cheng-Ping, Wu Guo-Jun, Wang Li-Li, Tan Yu-Rong
Respiratory Department, Xiangya Hospital, Central South University, Changsha, China.
Department of Basic Medicine, Xiangya School of Medicine, Central South University, Changsha, China.
Int Arch Allergy Immunol. 2019;179(1):43-52. doi: 10.1159/000495581. Epub 2019 Apr 3.
The aim of this study was to investigate the role of Notch-1 signaling through Notch-1 ligands on bronchial epithelial cells (BECs) in regulating the development of T helper 2 (Th2) lymphocytes after RSV infection.
Firstly, we analyzed the expression of cytokines and Notch-1 ligands in BECs by using real-time PCR. Then, RSV-infected BECs were co-cultured with CD4+ T cells in a transwell chamber for 48 h, and differentiation of T cells in the lower chamber was determined using flow cytometry and real-time PCR. JAG1 siRNA was then used to determine the effects of Jagged/Notch-1 signaling on the differentiation of Th2. An RSV-infected mouse model was also used to analyze the secretion of Th differentiation-associated cytokines in serum and lung tissues using ELISA, the histopathological changes using HE staining, and the expression of JAG1 and JAG2 in BECs.
The results showed that RSV promoted the expression of Th2-type cytokines and Jagged-1 and inhibited the expression of Jagged-2 in normal BECs. RSV-infected BECs induced Th2 differentiation. In addition, JAG1 downregulation inhibited the differentiation of Th2 and promoted differentiation of Th1. In the RSV-infected mouse model, the RSV titer, inflammation decreased with time. IL-4 and IL-17 increased on day 28 and 60, while IFNγ increased on day 7 and 28. Moreover, the expression of Jagged-1 increased and that of Jagged-2 decreased in BECs, which was consistent with IL-4 production in lung tissues.
Our data showed that BECs had the potential to promote the differentiation of Th2 lymphocytes through Jagged-1/Notch-1 signaling.
本研究旨在探讨Notch-1信号通过Notch-1配体作用于支气管上皮细胞（BECs）在呼吸道合胞病毒（RSV）感染后调节辅助性T细胞2（Th2）淋巴细胞发育中的作用。
首先，我们通过实时PCR分析BECs中细胞因子和Notch-1配体的表达。然后，将RSV感染的BECs与CD4+ T细胞在Transwell小室中共培养48小时，使用流式细胞术和实时PCR测定下室中T细胞的分化情况。接着使用JAG1 siRNA确定Jagged/Notch-1信号对Th2分化的影响。还使用RSV感染的小鼠模型，通过ELISA分析血清和肺组织中Th分化相关细胞因子的分泌情况，使用苏木精-伊红（HE）染色分析组织病理学变化，并分析BECs中JAG1和JAG2的表达情况。
结果显示，RSV促进正常BECs中Th2型细胞因子和Jagged-1的表达，并抑制Jagged-2的表达。RSV感染的BECs诱导Th2分化。此外，JAG1表达下调抑制Th2分化并促进Th1分化。在RSV感染的小鼠模型中，RSV滴度随时间下降，炎症减轻。IL-4和IL-17在第28天和第60天升高，而IFNγ在第7天和第28天升高。此外，BECs中Jagged-1的表达增加，Jagged-2的表达减少，这与肺组织中IL-4的产生一致。
我们的数据表明，BECs有潜力通过Jagged-1/Notch-1信号促进Th2淋巴细胞的分化。
