Xu Guo , Huifeng Dang , Wenji Huang , Zeinab Hassan , Shengran Yun , Yanan Lu , Yifan Liu , Junya Wang , Jun Zou
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引用次数: 0
摘要
白细胞介素(IL)20 是一种多功能细胞因子,在调节自身免疫性疾病、炎症和免疫反应方面发挥着重要作用。在鱼类中已经发现了 IL-20 的同源物。然而,由于缺乏抗体,鱼类 IL-20 的细胞来源和对感染反应的免疫学功能尚未完全定性。本研究生成了针对重组草鱼(Ctenopharyngodon idella)IL-20 蛋白的单克隆抗体(mAb),并通过免疫印迹、免疫荧光显微镜和流式细胞术对其进行了鉴定。结果表明,IL-20 mAb 能特异性识别在大肠杆菌细胞和 HEK293 细胞中表达的重组 IL-20 蛋白。利用共聚焦显微镜,在草鱼的头肾、鳃和肠道中发现了 IL-20+ 细胞,并在感染嗜水气单胞菌后诱导了这些细胞。此外,IL-20 蛋白主要由 CD3γδ T 细胞分泌,这些细胞主要位于鳃丝和肠粘膜。综上所述,我们的研究结果表明,鱼类的粘膜免疫需要产生 IL-20 的 T 细胞来抵御细菌感染。
IL-20 is produced by CD3γδ T cells and induced in the mucosal tissues of grass carp during infection with Aeromonas hydrophila
Interleukin (IL) 20 is a multifunctional cytokine and plays a vital role in regulating autoimmune diseases, inflammation, and immune responses. IL-20 homologs have been described in fish. However, due to the lack of antibodies, cellular sources and immunological functions of fish IL-20 in response to infections have not been fully characterized. In this study, a monoclonal antibody (mAb) was generated against the recombinant grass carp (Ctenopharyngodon idella) IL-20 protein and characterized by immunoblotting, immunofluorescent microscopy and flow cytometry. It was shown that the IL-20 mAb specifically recognized recombinant IL-20 proteins expressed in the E. coli cells and HEK293 cells. Using confocal microscopy, the IL-20+ cells were identified in the head kidney, gills and intestine of grass carp, and induced after infection with Aeromonas hydrophila. Moreover, the IL-20 protein was found to be secreted mainly by CD3γδ T cells which were located predominantly in the gill filaments and intestinal mucosa. Taken together, our results suggest that IL-20 producing T cells are required for the mucosal immunity against bacterial infection in fish.