Corthésy-Theulaz I, Porta N, Pringault E, Racine L, Bogdanova A, Kraehenbuhl J P, Blum A L, Michetti P
Division of Gastroenterology, Center Hospitalier Universitaire Vaudois, Lausanne, Switzerland.
Infect Immun. 1996 Sep;64(9):3827-32. doi: 10.1128/iai.64.9.3827-3832.1996.
Epithelial cells, which form tight polarized monolayers on porous substrates, constitute ideal model systems to study bacterial adhesion and invasion. The binding of Helicobacter pylori to the apical membrane of T84 cells, an epithelial cell line derived from a human colon carcinoma, was assessed biochemically and morphologically. Attachment was rapid, and binding remained constant over time, with a significant (P < 0.01, Mann-Whitney U test) ca. fourfold increase at pH 5.4 (76% +/- 22%) compared with pH 7.4 (18% +/- 7%). In contrast, adhesion of enteropathogenic Escherichia coli was not enhanced at pH 5.4. The transepithelial electrical resistance of the T84 cell monolayers was not affected by pH or by H. pylori. Following binding, H. pylori induced a reorganization of the brush border as reflected by actin condensation, facilitating the intimate association of the bacteria with the apical plasma membrane. H.pylori was not internalized, as shown by confocal microscopy. Some bacteria, found in deep invaginations of the apical membrane, were probably inaccessible to gentamicin, thus accounting for the observed tolerance to the antibiotic. These data provide the first evidence that an acidic environment favors Helicobacter adhesion and that binding is followed by survival of the survival of the bacteria in pockets of the apical membrane.
上皮细胞在多孔基质上形成紧密的极化单层,构成了研究细菌黏附和侵袭的理想模型系统。通过生化和形态学方法评估了幽门螺杆菌与人结肠癌来源的上皮细胞系T84细胞顶膜的结合情况。黏附迅速,且随着时间的推移结合保持恒定,在pH 5.4时(76%±22%)与pH 7.4时(18%±7%)相比有显著增加(P<0.01,Mann-Whitney U检验),约增加四倍。相比之下,pH 5.4时致病性大肠杆菌的黏附并未增强。T84细胞单层的跨上皮电阻不受pH或幽门螺杆菌的影响。结合后,幽门螺杆菌诱导刷状缘重组,表现为肌动蛋白凝聚,促进细菌与顶质膜紧密结合。共聚焦显微镜显示幽门螺杆菌未被内化。在顶膜深凹陷处发现的一些细菌可能无法接触到庆大霉素,因此解释了观察到的对该抗生素的耐受性。这些数据首次证明酸性环境有利于幽门螺杆菌黏附,且细菌黏附后能在顶膜凹陷处存活。