[Co-expression of CFA/I and CS6 of Enterotoxigenic Escherichia coli ( ETEC ) in Shigella flexneri 2a T32 Derivative Strain FWL01].

Ji-Ping Zheng, Ling-Chun Wang, Peng Wang, Gang Luo, Shu-Qin Li, Hai-Qing Duan, Cui-Fen Huang, Zhao-Shan Zhang
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Abstract

Among the known colonization factors of enterotoxigenic Escherichia coli (ETEC), CFA/I and CS6 (the common antigen in the CFA/IV fimbrial antigens ) are two of the most prevalent fimbriae found in clinical isolates but are never expressed by the same wild-type strains. In this study, CFA/I and CS6 of ETEC were co-expressed in Shigella flexneri 2a T32 derivative strain FWL01 by using a host-plasmid lethal balancing system based on asd gene. The results indicate that the recombinant plasmid carrying CFA/I and CS6 could be stably integrated in FWL01. Expression of the two antigens did not interfere the host growth. The results of immunofluorescence analysis showed that CFA/I and CS6 were localized on the surface of the strain FWL01. In Balb/c mice orally immunized with the recombinant strain, the immune responses against CFA/I and CS6 were observed. Those observations show the feasibility of a multivalent vaccine expressing different fimbrial antigens in attenuated Shigella flexneri.

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[产肠毒素大肠杆菌(ETEC) CFA/I和CS6在福氏志贺氏菌2a T32衍生菌株FWL01中的共表达]。
在已知的产肠毒素大肠杆菌(ETEC)定植因子中,CFA/I和CS6 (CFA/IV菌毛抗原中的共同抗原)是临床分离株中最常见的两种菌毛,但从未在同一种野生型菌株中表达。本研究利用基于asd基因的宿主-质粒致死平衡系统,在福氏志贺氏菌2a T32衍生菌株FWL01中共表达了CFA/I和CS6。结果表明,携带CFA/I和CS6的重组质粒能够稳定地整合到FWL01中。两种抗原的表达均不影响宿主的生长。免疫荧光分析结果显示,CFA/I和CS6定位于菌株FWL01的表面。用重组菌株口服免疫Balb/c小鼠,观察其对CFA/I和CS6的免疫应答。这些观察结果表明,在减毒的福氏志贺氏菌中表达不同纤维抗原的多价疫苗是可行的。
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