产肠毒素大肠杆菌感染IPEC-J2细胞后IL-17细胞因子表达变化及其功能初探

doi:10.11843/j.issn.0366-6964.2019.04.016

摘要/Abstract

摘要：

产肠毒素大肠杆菌（enterotoxigenic E.coli，ETEC）是引起新生和断奶仔猪腹泻（post-weaning diarrhea，PWD）的最重要的病原之一。ETEC进入肠道后与肠上皮细胞的相互作用既是该细菌致病的前提条件，也是激发宿主免疫反应的基础。本研究旨在分析ETEC感染猪肠上皮细胞后IL-17细胞因子表达变化，同时探索细胞因子在抵抗ETEC感染中的免疫防御机制。本研究采用猪肠上皮细胞系IPEC-J2和产肠毒素大肠杆菌标准株（C83901）为主要研究材料，通过RT-PCR、ELISA、免疫荧光及免疫印迹的方法分析了IL-17细胞因子及肠黏膜免疫防御相关基因在感染前后的变化。同时，进一步研究了IPEC-J2细胞中相关IL-17细胞因子刺激对肠黏膜防御相关基因的调控作用。结果表明，除IL-17D未检测到外，C83901能促进所有IL-17细胞因子mRNA的转录，尤其能显著上调IL-17A、IL-17C在基因和蛋白水平的表达。ETEC感染或者体外添加IL-17A/IL-17C有利于IPEC-J2细胞中黏蛋白（mucin-2）、紧密连接蛋白（claudin-1、claudin-2）及防御素pBD-2的表达。结果提示，ETEC感染后，肠上皮细胞通过调节IL-17细胞因子的表达进而增强肠黏膜屏障功能以抵抗该细菌的入侵。

关键词: IL-17, 产肠毒素大肠杆菌, 肠上皮细胞, 先天免疫, 宿主防御

Abstract:

Enterotoxigenic E. coli (ETEC) is one of the most important cause of neonatal and post-weaning diarrhea (PWD) in piglets. The interaction between ETEC and intestinal epithelial cells is the prerequisite of bacterial pathogenesis, as well as the first step of triggering host immune response. The aim of this study was to determine the expression profile and host defense mechanisms of IL-17 cytokine in porcine intestinal epithelial cells against ETEC infection. Firstly, we determined the expression of IL-17 cytokines and several mucosal defense genes by the application of RT-PCR, ELISA, immunohistochemistry (IHC) assays and Western Blotting in the porcine intestinal epithelial cell line IPEC-J2 infected with a F4⁺ETEC reference strain C83901. Later on, the regulating effect of IL-17 cytokine on the expression of above mucosal defense genes in IPEC-J2 cells was explored. The results showed that C83901 induced the mRNA expression of all IL-17 cytokines (IL-17D was not detected), particularly upregulated the expression of IL-17A and IL-17C both at the transcriptional and protein level. The upregulation of IL-17A and IL-17C in the protein level were confirmed by IHC and ELISA. The existence of IL-17A and/or IL-17C either induced by ETEC infection or supplemented artificially in IPEC-J2 cells can elicit the expression of host defense gene expression of mucin-2, claudin-1, claudin-2 and beta defensins-2. These results indicated that IL-17 induction in intestinal epithelial cells upon ETEC infection are beneficial for the mucosal host defense by promoting the intestinal barrier integrity.

Key words: interleukin-17, enterotoxigenic E. coli, intestinal epithelial cells, innate immune response, host defense

中图分类号:

S852.612

罗雨, 许佳, 张超颖, 蒋春燕, 何海健, 余建国, 章红兵. 产肠毒素大肠杆菌感染IPEC-J2细胞后IL-17细胞因子表达变化及其功能初探[J]. 畜牧兽医学报, 2019, 50(4): 830-839.

LUO Yu, XU Jia, ZHANG Chaoying, JIANG Chunyan, HE Haijian, YU Jianguo, ZHANG Hongbing. The Expression and Functional Analysis of IL-17 Cytokine Family in Enterotoxigenic E.coli Infected IPEC-J2 Cells[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(4): 830-839.

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