The Afa/Dr adhesins of diffusely adhering Escherichia coli stimulate interleukin-8 secretion, activate mitogen-activated protein kinases, and promote polymorphonuclear transepithelial migration in T84 polarized epithelial cells
The Afa/Dr adhesins of diffusely adhering Escherichia coli stimulate interleukin-8 secretion, activate mitogen-activated protein kinases, and promote polymorphonuclear transepithelial migration in T84 polarized epithelial cells
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DOI：
10.1128/iai.71.3.1068-1074.2003
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发表时间：
2003-03-01
影响因子：
3.1
通讯作者：
Hofman, P
中科院分区：
文献类型：
--
作者：
Bétis, F;Brest, P;Hofman, P
Afa/Dr弥散粘附性大肠杆菌（Afa/Dr DAEC）菌株可引起症状性尿路和肠道感染。迄今为止，尚未研究Afa/Dr DAEC菌株的体外促炎作用。在本研究中，我们使用汇合极化单层的肠细胞系T84评估上皮感染的后果Afa/博士DAEC菌株的促炎反应。将T84细胞单层与含有F1845菌毛粘附素的野生型Afa/Dr DAEC菌株C1845和含有Dr血凝素的菌株IH 11128孵育后，诱导多形性白细胞（PMNL）穿过上皮屏障迁移。用pSSS 1质粒转化大肠杆菌实验室菌株HB 101，产生Afa/Dr F1845粘附素。PMNL迁移与T84细胞的白细胞介素-8的基底外侧分泌相关，并且在上皮细胞与抗衰变加速因子（decay accelerating factor，ERF）抗体孵育后被废除，所述抗体识别ERF的短共识重复序列3结构域（单克隆抗体1H4）。此外，Afa/Dr DAEC菌株诱导几种T84蛋白的酪氨酸磷酸化，并激活促分裂原活化蛋白激酶（ERK 1/2促分裂原活化蛋白、P38和Jun-C激酶）。这些数据首次证明，在体外，Afa/Dr DAEC菌株在肠上皮细胞中产生促炎信号。
Afa/Dr diffusely adhering Escherichia coli (Afa/Dr DAEC) strains cause symptomatic urinary tract and intestinal infections. The proinflammatory effects of Afa/Dr DAEC strains in vitro have been not investigated to date. In the present study, we used confluent polarized monolayers of intestinal cell line T84 to evaluate the consequences of epithelial infection by Afa/Dr DAEC strains in terms of proinflammatory response. Polymorphonuclear leukocyte (PMNL) migration across the epithelial barrier was induced after incubation of the T84 monolayers with the wild-type Afa/Dr DAEC strain C1845 harboring the fimbrial F1845 adhesin and strain IH11128 harboring the Dr hemagglutinin, and the E. coli laboratory strain HB101 was transformed with the pSSS1 plasmid, producing Afa/Dr F1845 adhesin. PMNL migrations were correlated with a basolateral secretion of interleukin-8 by T84 cells and were abolished after incubation of epithelial cells with an anti-decay accelerating factor (DAF) antibody that recognized the short consensus repeat 3 domain of DAF (monoclonal antibody 1H4). Moreover, Afa/Dr DAEC strains induced tyrosine phosphorylation of several T84 proteins and activated the mitogen-activated protein kinases (ERK1/2 mitogen-activated protein, P38, and Jun-C kinases). These data demonstrated for the first time that, in vitro, Afa/Dr DAEC strains exert a proinflammatory signal in intestinal epithelial cells.