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在乳球菌 NZ9000 中表达 EspA 及其体内外免疫效果的检测。

Expression of EspA in Lactococcus lactis NZ9000 and the detection of its immune effect in vivo and vitro.

机构信息

State Key Laboratory of Microbial Technology, Shandong University, Jinan, Shandong, People's Republic of China.

出版信息

Immunopharmacol Immunotoxicol. 2010 Mar;32(1):133-40. doi: 10.3109/08923970903207083.

Abstract

Enterhemorrhagic Escherichia coli (EPEC), an important cause of severe infantile diarrheal disease in many parts of the developing world, produced several recently described virulence determinations. Several of its virulence factors are secreted by type III secretion including EspA, which forms filamentous structures on bacterial surface bridging to the host cell's surface. These structures on bacterial surfaces may deliver other virulence factors directly into the host cell from EHEC. In this study, EspA was expressed in Lactococcus lactis NZ9000 (L. lactis NZ9000). BALB/C mice were immunized by recombinant EspA, and mice sera were assayed for the disruption of E. coli O157:H7 interaction with the host cell. BALB/C mice which were immunized with recombinant EspA produced specific antibody titers, and the difference between the control group and the immune group is marked (P<0.05). The polyclonal mice antisera blocked E. coli O157:H7-induced host cell actin rearrangement and could label E. coli O157:H7 EspA filaments in vitro. Our present findings suggest that L. lactis may be a good candidate to produce oral vaccine, and thus entertains the possibility of developing EspA-oral vaccine in the defense of E. coli O157:H7 infection.

摘要

产志贺样毒素大肠杆菌(Enterohaemorrhagic Escherichia coli,EHEC)是发展中国家许多地区导致严重婴幼儿腹泻病的重要病因,它产生了几种最近描述的毒力决定因素。其几种毒力因子通过 III 型分泌系统分泌,包括 EspA,EspA 在细菌表面形成丝状结构,与宿主细胞表面桥接。这些细菌表面的结构可能将其他毒力因子直接从 EHEC 递送到宿主细胞中。在这项研究中,EspA 在乳球菌 lactis NZ9000(L. lactis NZ9000)中表达。BALB/C 小鼠用重组 EspA 免疫,检测小鼠血清中破坏大肠杆菌 O157:H7 与宿主细胞相互作用的能力。用重组 EspA 免疫的 BALB/C 小鼠产生了特异性抗体滴度,免疫组和对照组之间的差异非常显著(P<0.05)。多克隆小鼠抗血清可阻断大肠杆菌 O157:H7 诱导的宿主细胞肌动蛋白重排,并可在体外标记大肠杆菌 O157:H7 EspA 丝。我们目前的研究结果表明,乳球菌可能是生产口服疫苗的良好候选者,因此有可能开发 EspA-口服疫苗来防御大肠杆菌 O157:H7 感染。

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