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用表达衣原体低钙反应蛋白E(LcrE)的DNA载体接种疫苗以预防肺炎衣原体感染。

Vaccination with DNA vector expressing chlamydial low calcium response protein E (LcrE) against Chlamydophila pneumoniae infection.

作者信息

Faludi Ildikó, Szabó Ágnes Míra

机构信息

Department of Medical Microbiology and Immunobiology, University of Szeged, Szeged, Hungary.

出版信息

Acta Microbiol Immunol Hung. 2011 Jun;58(2):123-34. doi: 10.1556/AMicr.58.2011.2.5.

Abstract

Chlamydophila pneumoniae is an obligate intracellular human pathogen, which causes acute respiratory tract infections and can also cause chronic infections. C. pneumoniae possess type III secretion system (TTSS), which allows them to secrete effector molecules into the inclusion membrane and the host cell cytosol. Low calcium response protein E (LcrE) is a part of TTSS. The gene of LcrE in a 6His-tagged form was cloned from C. pneumoniae CWL029, expressed and purified from Escherichia coli using the HIS-select TALON CellThru Resin, this gene was also cloned into a eukaryotic expression vector (pΔRC). One group of BALB/c mice received an intramuscular pΔRC inoculation then the mice were immunized with purified LcrE protein; the second group of mice was immunized two times with the recombinant plasmid (pΔRCLcrE), and the third group was primed with pΔRCLcrE inoculation then boosted with LcrE protein. LcrE-specific antibody response was induced by DNA immunization with a shift towards Th1 isotype pattern compared to protein-immunization, this shifting pattern was observed in plasmid primed then protein-boosted animals. DNA immunization given as a priming and followed by a protein booster significantly reduced the number of viable bacteria in the lungs after challenge with C. pneumoniae. These results confirm that immunization with pΔRCLcrE can be an effective part of a vaccination schedule against C. pneumoniae.

摘要

肺炎衣原体是一种专性胞内人类病原体,可引起急性呼吸道感染,也能导致慢性感染。肺炎衣原体拥有III型分泌系统(TTSS),这使其能够将效应分子分泌到包涵体膜和宿主细胞胞质溶胶中。低钙反应蛋白E(LcrE)是TTSS的一部分。以6His标签形式存在的LcrE基因从肺炎衣原体CWL029中克隆出来,利用HIS-select TALON CellThru树脂在大肠杆菌中进行表达和纯化,该基因也被克隆到真核表达载体(pΔRC)中。一组BALB/c小鼠接受肌肉注射pΔRC,然后用纯化的LcrE蛋白进行免疫;第二组小鼠用重组质粒(pΔRCLcrE)免疫两次,第三组先用pΔRCLcrE接种进行初免,然后用LcrE蛋白进行加强免疫。与蛋白质免疫相比,DNA免疫诱导了LcrE特异性抗体反应,并向Th1同种型模式转变,这种转变模式在质粒初免然后蛋白质加强免疫的动物中观察到。以DNA免疫作为初免随后进行蛋白质加强免疫,在用肺炎衣原体攻击后,显著减少了肺中活细菌的数量。这些结果证实,用pΔRCLcrE进行免疫可以成为抗肺炎衣原体疫苗接种计划的有效组成部分。

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