Sampaio Suely C F, Gomes Tânia A T, Pichon Christophe, du Merle Laurence, Guadagnini Stéphanie, Abe Cecilia M, Sampaio Jorge L M, Le Bouguénec Chantal
Departamento de Microbiologia, Imunologia e Parasitologia, Universidade Federal de São Paulo, Escola Paulista de Medicina, Rua Botucatu 862, 3 degrees andar, 04023-062 São Paulo, Brazil.
Infect Immun. 2009 Oct;77(10):4406-13. doi: 10.1128/IAI.00177-09. Epub 2009 Jul 20.
The ability of some typical enteropathogenic Escherichia coli (EPEC) strains to adhere to, invade, and increase interleukin-8 (IL-8) production in intestinal epithelial cells in vitro has been demonstrated. However, few studies regarding these aspects have been performed with atypical EPEC (aEPEC) strains, which are emerging enteropathogens in Brazil. In this study, we evaluated a selected aEPEC strain (1711-4) of serotype O51:H40, the most prevalent aEPEC serotype in Brazil, in regard to its ability to adhere to and invade Caco-2 and T84 cells and to elicit IL-8 production in Caco-2 cells. The role of flagella in aEPEC 1711-4 adhesion, invasion, and IL-8 production was investigated by performing the same experiments with an isogenic aEPEC mutant unable to produce flagellin (FliC), the flagellum protein subunit. We demonstrated that this mutant (fliC mutant) had a marked decrease in the ability to adhere to T84 cells and invade both T84 and Caco-2 cells in gentamicin protection assays and by transmission electron microscopy. In addition, the aEPEC 1711-4 fliC mutant had a reduced ability to stimulate IL-8 production by Caco-2 cells in early (3-h) but not in late (24-h) infections. Our findings demonstrate that flagella of aEPEC 1711-4 are required for efficient adhesion, invasion, and early but not late IL-8 production in intestinal epithelial cells in vitro.
一些典型的肠致病性大肠杆菌(EPEC)菌株在体外对肠道上皮细胞的黏附、侵袭以及增加白细胞介素-8(IL-8)产生的能力已得到证实。然而,针对这些方面,很少有研究是使用非典型EPEC(aEPEC)菌株进行的,而aEPEC菌株在巴西是新出现的肠道病原体。在本研究中,我们评估了一株选定的血清型为O51:H40的aEPEC菌株(1711-4),它是巴西最常见的aEPEC血清型,研究其对Caco-2和T84细胞的黏附、侵袭能力以及在Caco-2细胞中引发IL-8产生的能力。通过对一株无法产生鞭毛蛋白(FliC,鞭毛的蛋白质亚基)的同基因aEPEC突变体进行相同实验,研究了鞭毛在aEPEC 1711-4黏附、侵袭和IL-8产生中的作用。我们证明,在庆大霉素保护试验和透射电子显微镜观察中,该突变体(fliC突变体)对T84细胞的黏附能力以及对T84和Caco-2细胞的侵袭能力均显著下降。此外,aEPEC 1711-4 fliC突变体在早期(3小时)感染时刺激Caco-2细胞产生IL-8的能力降低,但在晚期(24小时)感染时未降低。我们的研究结果表明,aEPEC 1711-4的鞭毛对于体外肠道上皮细胞的有效黏附、侵袭以及早期而非晚期IL-8的产生是必需的。