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鉴定钩端螺旋体蛋白,这些蛋白在仓鼠中提供部分保护,防止致死性挑战钩端螺旋体 interrogans。

Characterization of leptospiral proteins that afford partial protection in hamsters against lethal challenge with Leptospira interrogans.

机构信息

Centro de Biotecnologia, Instituto Butantan, Avenida Vital Brazil 1500, 05503-900 São Paulo, SP, Brazil.

Laboratório de Zoonoses Bacterianas do VPS, Faculdade de Medicina Veterinária e Zootecnia, USP, Avenida Prof. Dr Orlando Marques de Paiva 87, 05508-270 São Paulo, SP, Brazil.

出版信息

J Med Microbiol. 2010 Sep;59(Pt 9):1005-1015. doi: 10.1099/jmm.0.021485-0. Epub 2010 Jun 17.

Abstract

Leptospirosis is a worldwide zoonosis caused by pathogenic Leptospira. The whole-genome sequence of Leptospira interrogans serovar Copenhageni together with bioinformatic tools allow us to search for novel antigen candidates suitable for improved vaccines against leptospirosis. This study focused on three genes encoding conserved hypothetical proteins predicted to be exported to the outer membrane. The genes were amplified by PCR from six predominant pathogenic serovars in Brazil. The genes were cloned and expressed in Escherichia coli strain BL21-SI using the expression vector pDEST17. The recombinant proteins tagged with N-terminal 6xHis were purified by metal-charged chromatography. The proteins were recognized by antibodies present in sera from hamsters that were experimentally infected. Immunization of hamsters followed by challenge with a lethal dose of a virulent strain of Leptospira showed that the recombinant protein rLIC12730 afforded statistically significant protection to animals (44 %), followed by rLIC10494 (40 %) and rLIC12922 (30 %). Immunization with these proteins produced an increase in antibody titres during subsequent boosters, suggesting the involvement of a T-helper 2 response. Although more studies are needed, these data suggest that rLIC12730 and rLIC10494 are promising candidates for a multivalent vaccine for the prevention of leptospirosis.

摘要

钩端螺旋体病是一种由致病性钩端螺旋体引起的世界性动物传染病。钩端螺旋体 interrogans 血清型哥本哈根的全基因组序列和生物信息学工具使我们能够寻找适合改良钩端螺旋体病疫苗的新型抗原候选物。本研究集中于三个编码保守假设蛋白的基因,这些蛋白被预测为可向外膜输出。从巴西六种主要致病性血清型中通过 PCR 扩增这些基因。使用表达载体 pDEST17 将基因克隆并在大肠杆菌 BL21-SI 中表达。重组蛋白带有 N 端 6xHis 标签,通过金属离子亲和层析纯化。这些蛋白被实验感染的仓鼠血清中的抗体识别。用重组蛋白 rLIC12730 免疫仓鼠,然后用致死剂量的毒力菌株挑战,结果表明重组蛋白 rLIC12730 使动物(44%)获得了统计学上显著的保护,其次是 rLIC10494(40%)和 rLIC12922(30%)。这些蛋白的免疫接种在随后的加强免疫中增加了抗体滴度,表明存在辅助性 T 细胞 2 反应。尽管还需要更多的研究,但这些数据表明 rLIC12730 和 rLIC10494 是预防钩端螺旋体病的多价疫苗的有前途的候选物。

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