对军团病的保护作用:重组嗜肺军团菌鞭毛蛋白A在BALB/c小鼠模型中诱导对菌血症的保护性免疫

IF 1.2 Q2 Biochemistry, Genetics and Molecular Biology Journal of Molecular Microbiology and Biotechnology Pub Date : 2017-01-01 Epub Date: 2017-04-26 DOI:10.1159/000460295
Ashraf Mohabati Mobarez, Roya Ahmadrajabi, Nima Khoramabadi, Ali-Hatef Salmanian
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引用次数: 6

摘要

为了研究重组A型鞭毛蛋白(FLA)的免疫保护作用,将嗜肺军团菌血清1群Paris菌株flaA基因克隆到pET28a(+)中。重组FLA (rFLA)在大肠杆菌BL21 (DE3)中过表达,并通过Ni2+交换层析纯化。用20 μg rFLA免疫6 ~ 8周龄雌性BALB/c。未免疫小鼠和静脉注射亚致死剂量嗜肺乳杆菌的小鼠分别作为阴性对照和阳性对照。雌性BALB/c小鼠经rFLA免疫后血清抗体应答显著。血清和脾细胞培养物中IFN-γ、IL-12和TNF-α的产生以及抗原特异性脾细胞的增殖表明,在rfla免疫小鼠中存在强烈的先天和适应性细胞介导的免疫反应。在10天的随访调查中,嗜肺乳杆菌血清组1 (Paris菌株)静脉致死性攻毒显示,60%接种rFLA的小鼠存活。这些结果表明,嗜肺乳杆菌的rFLA可引起强烈的先天和适应性免疫反应,并提示对嗜肺乳杆菌致命攻击的长期免疫的可能性。
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Protection against Legionnaire's Disease: Recombinant Flagellin A of Legionella pneumophila Can Induce Protective Immunity against Bacteremia in a BALB/c Murine Model.
To investigate the immunoprotective effects of the recombinant type A flagellin (FLA), the flaA gene of Legionella pneumophila serogroup 1 strain Paris was cloned into pET28a(+). Recombinant FLA (rFLA) was overexpressed in E. coli BL21 (DE3) and purified by Ni2+ exchange chromatography. Female BALB/c aged 6-8 weeks were immunized with 20 μg of rFLA. Nonimmunized mice along with mice inoculated with a sublethal dose of live L. pneumophila intravenously were considered as negative and positive controls, respectively. A significant serum antibody response was observed in female BALB/c mice immunized with rFLA. Production of IFN-γ and IL-12, and TNF-α in the serum and the splenocyte cultures, and antigen-specific splenocyte proliferation suggested a strong innate and adaptive cell-mediated immunity response in rFLA-immunized mice. Intravenous lethal challenge with L. pneumophila serogroup 1 (strain Paris) showed that 60% of mice immunized with rFLA survived in a 10-day follow-up survey. These results show that rFLA from L. pneumophila can elicit strong innate and adaptive immune responses and suggest the possibility of a long-term immunity against lethal challenge with L. pneumophila.
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来源期刊
Journal of Molecular Microbiology and Biotechnology
Journal of Molecular Microbiology and Biotechnology 生物-生物工程与应用微生物
CiteScore
3.90
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0.00%
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0
审稿时长
>12 weeks
期刊介绍: We are entering a new and exciting era of microbiological study and application. Recent advances in the now established disciplines of genomics, proteomics and bioinformatics, together with extensive cooperation between academic and industrial concerns have brought about an integration of basic and applied microbiology as never before.
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Contents Front & Back Matter The Life Cycle of Dictyostelium discoideum Is Accelerated via MAP Kinase Cascade by a Culture Extract Produced by a Synthetic Microbial Consortium A Riboflavin Transporter in Bdellovibrio exovorous JSS Front & Back Matter
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