Characterization of a porcine intestinal epithelial cell line for in vitro studies of microbial pathogenesis in swine
Characterization of a porcine intestinal epithelial cell line for in vitro studies of microbial pathogenesis in swine
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DOI：
10.1007/s00418-005-0067-z
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发表时间：
2006-03-01
影响因子：
2.3
通讯作者：
Wieler, LH
中科院分区：
文献类型：
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作者：
Schierack, P;Nordhoff, M;Wieler, LH
由于缺乏相关的猪细胞系，对猪发病机制的体外研究一直受到阻碍。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适当的猪源细胞系。我们利用多种方法对猪肠上皮细胞系IPEC-J2进行了表征，以评估该细胞系作为体外感染模型的有效性。电镜和组织化学染色显示细胞呈肠细胞样，有微绒毛、紧密连接和糖萼结合的粘蛋白。单层膜的功能完整性是通过上皮电阻（TEER）测量来确定的。选择共生细菌和重要的细菌病原体进行研究，主要是基于它们不同的感染机制：专性细胞外大肠杆菌、兼性细胞内沙门氏菌和专性细胞内衣原体。我们确定了细菌在宿主细胞上和宿主细胞内的定植和增殖，并监测了宿主细胞的反应。我们验证了编码细胞因子IL-1 α、-6、-7、-8、-18、tnf - α和GM-CSF的mrna的表达，但没有编码tgf - β或MCP-1。沙门氏菌侵染后IL-8蛋白表达增强。我们认为IPEC-J2细胞系为猪肠道病原体-宿主细胞相互作用提供了一个相关的体外模型系统。
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-1 alpha, -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.