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一种用于猪微生物发病机制体外研究的猪肠道上皮细胞系的特性分析。

Characterization of a porcine intestinal epithelial cell line for in vitro studies of microbial pathogenesis in swine.

作者信息

Schierack Peter, Nordhoff Marcel, Pollmann Marion, Weyrauch Karl Dietrich, Amasheh Salah, Lodemann Ulrike, Jores Jörg, Tachu Babila, Kleta Sylvia, Blikslager Anthony, Tedin Karsten, Wieler Lothar H

机构信息

Institut für Mikrobiologie und Tierseuchen, Freie Universität Berlin, Berlin, Germany.

出版信息

Histochem Cell Biol. 2006 Mar;125(3):293-305. doi: 10.1007/s00418-005-0067-z. Epub 2005 Oct 8.

Abstract

In vitro studies on the pathogenesis in swine have been hampered by the lack of relevant porcine cell lines. Since many bacterial infections are swine-specific, studies on pathogenic mechanisms require appropriate cell lines of porcine origin. We have characterized the permanent porcine intestinal epithelial cell line, IPEC-J2, using a variety of methods in order to assess the usefulness of this cell line as an in vitro infection model. Electron microscopic analyses and histochemical staining revealed the cells to be enterocyte-like with microvilli, tight junctions and glycocalyx-bound mucin. The functional integrity of monolayers was determined by transepithelial electrical resistance (TEER) measurements. Both commensal bacteria and important bacterial pathogens were chosen for study based on their principally different infection mechanisms: obligate extracellular Escherichia coli, facultative intracellular Salmonella and obligate intracellular Chlamydia. We determined the colonization and proliferation of the bacteria on and within the host cells and monitored the host cell response. We verified the expression of mRNAs encoding the cytokines IL-1alpha, -6, -7, -8, -18, TNF-alpha and GM-CSF, but not TGF-beta or MCP-1. IL-8 protein expression was enhanced by Salmonella invasion. We conclude that the IPEC-J2 cell line provides a relevant in vitro model system for porcine intestinal pathogen-host cell interactions.

摘要

由于缺乏相关的猪细胞系,猪发病机制的体外研究受到了阻碍。由于许多细菌感染是猪特有的,因此对致病机制的研究需要合适的猪源细胞系。我们使用多种方法对永久性猪肠上皮细胞系IPEC-J2进行了表征,以评估该细胞系作为体外感染模型的实用性。电子显微镜分析和组织化学染色显示,这些细胞呈肠上皮细胞样,具有微绒毛、紧密连接和糖萼结合的粘蛋白。单层细胞的功能完整性通过跨上皮电阻(TEER)测量来确定。基于共生细菌和重要细菌病原体主要不同的感染机制,选择了专性胞外大肠杆菌、兼性胞内沙门氏菌和专性胞内衣原体进行研究。我们确定了细菌在宿主细胞上和细胞内的定殖和增殖,并监测了宿主细胞的反应。我们验证了编码细胞因子IL-1α、-6、-7、-8、-18、TNF-α和GM-CSF的mRNA的表达,但未验证TGF-β或MCP-1的表达。沙门氏菌入侵增强了IL-8蛋白的表达。我们得出结论,IPEC-J2细胞系为猪肠道病原体与宿主细胞相互作用提供了一个相关的体外模型系统。

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