Neves Bianca C, Shaw Robert K, Frankel Gad, Knutton Stuart
Centre for Molecular Microbiology and Infection, Department of Biological Sciences, Imperial College London, London SW7 2AZ, UK.
Infect Immun. 2003 Apr;71(4):2262-5. doi: 10.1128/IAI.71.4.2262-2265.2003.
Enteropathogenic Escherichia coli (EPEC) and enterohemorrhagic E. coli (EHEC) possess a filamentous type III secretion system (TTSS) employed to deliver effector proteins into host cells. EspA is a type III secreted protein which forms the filamentous extension to the TTSS and which interacts with host cells during early stages of attaching and effacing (A/E) lesion formation. By immunofluorescence, a polyclonal antibody previously raised to EspA from EPEC strain E2348/69 (O127:H6) stained approximately 12-nm-diameter EspA filaments produced by this strain but did not stain similar filaments produced by EHEC serotype O157:H7. Similarly, an antibody that we subsequently raised to EHEC strain 85-170 (O157:H7) EspA stained approximately 12-nm-diameter EspA filaments produced by strain 85-170 but did not stain E2348/69 EspA filaments. Given such heterogeneity between EPEC and EHEC EspA filaments, we examined polymorphisms of functional EspA filaments among different EPEC and EHEC serotypes. With use of the EPEC EspA antiserum, EspA filaments were observed only with EPEC serotypes O127:H6 and O55:H6, serotypes which encode an identical EspA protein. When stained with the EHEC EspA antiserum, EspA filaments were detected only on EHEC strains belonging to serotype O157:H7; the EHEC antiserum did, however, stain EspA filaments produced by the closely related EPEC serotype O55:H7 but not filaments of any other EPEC serotype tested. Such polymorphisms among functional EspA filaments of EPEC and EHEC would be expected to have important implications for the development of broad-range EspA-based vaccines.
肠致病性大肠杆菌(EPEC)和肠出血性大肠杆菌(EHEC)拥有一种丝状III型分泌系统(TTSS),用于将效应蛋白输送到宿主细胞中。EspA是一种III型分泌蛋白,它形成TTSS的丝状延伸部分,并在黏附和损伤形成(A/E)病变的早期阶段与宿主细胞相互作用。通过免疫荧光法,先前针对EPEC菌株E2348/69(O127:H6)的EspA产生的多克隆抗体可对该菌株产生的直径约为12纳米的EspA丝进行染色,但不能对EHEC血清型O157:H7产生的类似丝进行染色。同样,我们随后针对EHEC菌株85-170(O157:H7)的EspA产生的抗体可对85-170菌株产生的直径约为12纳米的EspA丝进行染色,但不能对E2348/69的EspA丝进行染色。鉴于EPEC和EHEC的EspA丝之间存在这种异质性,我们研究了不同EPEC和EHEC血清型中功能性EspA丝的多态性。使用EPEC EspA抗血清时,仅在EPEC血清型O127:H6和O55:H6中观察到EspA丝,这两种血清型编码相同的EspA蛋白。用EHEC EspA抗血清染色时,仅在属于血清型O157:H