Mohabati Mobarez Ashraf, Ahmadrajabi Roya, Khoramabadi Nima, Salmanian Ali-Hatef
Department of Bacteriology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
J Mol Microbiol Biotechnol. 2017;27(2):110-116. doi: 10.1159/000460295. Epub 2017 Apr 26.
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.
为研究重组A型鞭毛蛋白(FLA)的免疫保护作用,将嗜肺军团菌血清1型巴黎菌株的flaA基因克隆至pET28a(+)载体。重组FLA(rFLA)在大肠杆菌BL21(DE3)中过量表达,并通过Ni2+交换层析进行纯化。用20μg rFLA对6 - 8周龄雌性BALB/c小鼠进行免疫。未免疫小鼠以及静脉接种亚致死剂量活嗜肺军团菌的小鼠分别作为阴性和阳性对照。在用rFLA免疫的雌性BALB/c小鼠中观察到显著的血清抗体反应。血清和脾细胞培养物中IFN-γ、IL-12以及TNF-α的产生,以及抗原特异性脾细胞增殖表明,在rFLA免疫的小鼠中存在强烈的先天性和适应性细胞介导免疫反应。用嗜肺军团菌血清1型(巴黎菌株)进行静脉致死性攻击显示,在为期10天的随访调查中,用rFLA免疫的小鼠有60%存活。这些结果表明,来自嗜肺军团菌的rFLA可引发强烈的先天性和适应性免疫反应,并提示存在针对嗜肺军团菌致死性攻击产生长期免疫的可能性。