Yamamoto Denise, Hernandes Rodrigo T, Blanco Miguel, Greune Lilo, Schmidt M Alexander, Carneiro Sylvia M, Dahbi Ghizlane, Blanco Jesús E, Mora Azucena, Blanco Jorge, Gomes Tânia A T
Universidade Federal de São Paulo, Rua Botucatu, 862, 3o andar, Vila Clementino, São Paulo, CEP 04023-062, Brazil.
BMC Microbiol. 2009 Jul 21;9:146. doi: 10.1186/1471-2180-9-146.
Enteropathogenic Escherichia coli (EPEC) produce attaching/effacing (A/E) lesions on eukaryotic cells mediated by the outer membrane adhesin intimin. EPEC are sub-grouped into typical (tEPEC) and atypical (aEPEC). We have recently demonstrated that aEPEC strain 1551-2 (serotype O non-typable, non-motile) invades HeLa cells by a process dependent on the expression of intimin sub-type omicron. In this study, we evaluated whether aEPEC strains expressing other intimin sub-types are also invasive using the quantitative gentamicin protection assay. We also evaluated whether aEPEC invade differentiated intestinal T84 cells.
Five of six strains invaded HeLa and T84 cells in a range of 13.3%-20.9% and 5.8%-17.8%, respectively, of the total cell-associated bacteria. The strains studied were significantly more invasive than prototype tEPEC strain E2348/69 (1.4% and 0.5% in HeLa and T84 cells, respectively). Invasiveness was confirmed by transmission electron microscopy. We also showed that invasion of HeLa cells by aEPEC 1551-2 depended on actin filaments, but not on microtubules. In addition, disruption of tight junctions enhanced its invasion efficiency in T84 cells, suggesting preferential invasion via a non-differentiated surface.
Some aEPEC strains may invade intestinal cells in vitro with varying efficiencies and independently of the intimin sub-type.
肠致病性大肠杆菌(EPEC)通过外膜黏附素紧密黏附素介导,在真核细胞上产生黏附/抹消(A/E)损伤。EPEC可分为典型(tEPEC)和非典型(aEPEC)两类。我们最近证明,aEPEC菌株1551-2(血清型O不可分型、无动力)通过依赖于紧密黏附素亚型omicron表达的过程侵入HeLa细胞。在本研究中,我们使用定量庆大霉素保护试验评估表达其他紧密黏附素亚型的aEPEC菌株是否也具有侵袭性。我们还评估了aEPEC是否侵入分化的肠道T84细胞。
六株菌株中有五株分别以占总细胞相关细菌的13.3%-20.9%和5.8%-17.8%的比例侵入HeLa细胞和T84细胞。所研究的菌株比原型tEPEC菌株E2348/69的侵袭性显著更高(在HeLa细胞和T84细胞中分别为1.4%和0.5%)。通过透射电子显微镜证实了侵袭性。我们还表明,aEPEC 1551-2对HeLa细胞的侵袭依赖于肌动蛋白丝,但不依赖于微管。此外,紧密连接的破坏增强了其在T84细胞中的侵袭效率,表明通过未分化表面的优先侵袭。
一些aEPEC菌株可能在体外以不同效率侵入肠道细胞,且与紧密黏附素亚型无关。