Beinke C, Laarmann S, Wachter C, Karch H, Greune L, Schmidt M A
Institut für Infektiologie, Zentrum für Molekularbiologie der Entzündung, Westfälische Wilhelms-Universität, Münster, Germany.
Infect Immun. 1998 Feb;66(2):528-39. doi: 10.1128/IAI.66.2.528-539.1998.
Recent epidemiological studies indicate that Escherichia coli strains which exhibit the diffuse-adherence phenotype (DAEC strains) represent a potential cause of diarrhea in infants. We investigated the interaction of DAEC strains isolated from diarrhea patients in Brazil and in Germany with epithelial cells in tissue culture. The investigated strains were identified as DAEC strains by (i) their attachment pattern, (ii) presence of genes associated with the Dr family of adhesins, and (iii) lack of genetic markers for other diarrhea-associated E. coli categories. Several clinical DAEC isolates were shown to secrete similar patterns of proteins into tissue culture medium. Protein secretion was found to be regulated by environmental parameters, namely, medium, temperature, pH, and iron concentration. DAEC strains secreting these proteins induced accumulation of actin and tyrosine-phosphorylated proteins at sites of bacterial attachment, leading to the formation of pedestals and/or extended surface structures. These changes were phenotypically similar to the attaching and effacing (A/E) lesions observed with enteropathogenic and some enterohemorrhagic E. coli strains carrying the locus of enterocyte effacement (LEE) pathogenicity island. Proteins homologous to the EspA, EspB, and EspD proteins, necessary for signal transduction events inducing A/E lesions, were identified by sequence analysis and cross-reaction of specific antibodies. However, initially nonadhering strains secreting these proteins induced signal transduction events only after prolonged infection. These results indicate that secretion of the Esp proteins alone is not sufficient for efficient signal transduction. This study further shows that some DAEC strains are likely to contain a homolog(s) of the LEE locus which may contribute to the pathogenic potential of DAEC.
近期的流行病学研究表明,呈现弥散黏附表型的大肠杆菌菌株(DAEC菌株)是婴儿腹泻的一个潜在病因。我们研究了从巴西和德国腹泻患者中分离出的DAEC菌株与组织培养中的上皮细胞之间的相互作用。通过以下方法将所研究的菌株鉴定为DAEC菌株:(i)它们的黏附模式;(ii)存在与Dr黏附素家族相关的基因;(iii)缺乏其他腹泻相关大肠杆菌类别的遗传标记。数种临床DAEC分离株被证明能向组织培养基中分泌相似模式的蛋白质。发现蛋白质分泌受环境参数调节,即培养基、温度、pH值和铁浓度。分泌这些蛋白质的DAEC菌株在细菌黏附位点诱导肌动蛋白和酪氨酸磷酸化蛋白积累,导致基座和/或扩展表面结构的形成。这些变化在表型上类似于携带肠细胞脱落位点(LEE)致病岛的肠致病性和一些肠出血性大肠杆菌菌株所观察到的黏附与脱落(A/E)损伤。通过序列分析和特异性抗体的交叉反应,鉴定出了与诱导A/E损伤的信号转导事件所必需的EspA、EspB和EspD蛋白同源的蛋白质。然而,最初不黏附的分泌这些蛋白质的菌株仅在长时间感染后才诱导信号转导事件。这些结果表明,仅Esp蛋白的分泌不足以实现有效的信号转导。这项研究进一步表明,一些DAEC菌株可能含有LEE位点的同源物,这可能有助于DAEC的致病潜力。