Construction of recombinant Mip-FlaA dominant epitope vaccine against Legionella pneumophila and evaluation of the immun
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ORIGINAL ARTICLE
Construction of recombinant Mip-FlaA dominant epitope vaccine against Legionella pneumophila and evaluation of the immunogenicity and protective immunity Jinlei He1 • Junrong Zhang1 • Yanxia He1,2 • Fan Huang3 • Jiao Li1 Qiwei Chen1 • Dali Chen1 • Jianping Chen1,4
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Ó Springer Science+Business Media New York 2015
Abstract Legionnaires’ disease, a kind of systemic disease with pneumonia as the main manifestation, is caused by Legionella pneumophila (L. pneumophila). In order to prevent the disease, we optimized Mip and FlaA, the virulence factors of L. pneumophila, to design recombinant Mip-FlaA dominant epitope vaccine against the pathogen. Firstly, the coding sequences of mip and flaA were optimized by DNAStar software and Expasy protein analysis system, and then, the tertiary structure and function of recombinant MipFlaA were predicted by PHYRE2 Protein Fold Recognition Server. After that, the optimized mip, flaA and mip-flaA were cloned, expressed and purified, and the proteins were used as dominant epitope vaccines to immunize BABL/c mice. Moreover, the IgG titers, histological changes in lung and the level of TNF-a, IFN-c, IL-6 and IL-1b were detected to reflect the immunogenicity and protective immunity of the vaccines. The results of SDS-PAGE and Western blot proved the recombinant Mip-FlaA was successfully expressed. ELISA results of IgG titers and these cytokines showed Mip-
& Jianping Chen [email protected] 1
Department of Parasitology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China
2
Department of Pathogen Biology and Immunology, Medical College, Hubei University for Nationalities, Enshi 445000, China
3
First Department of Hepatobiliary Surgery, Qinghai University Affiliated Hospital, Medical College of Qinghai University, Xining 810016, China
4
Animal Disease Prevention and Food Safety Key Laboratory of Sichuan Province, School of Life Science, Sichuan University, Chengdu 610041, China
FlaA group was capable to induce the strongest immune response, compared to PBS, Mip and FlaA groups. In addition, histopathology analysis demonstrated the mice immunized with Mip-FlaA showed better immune protection. Therefore, the work indicated that the above-described biological tools were useful in optimization of epitope vaccine. Antigenic characterization and immune protection of recombinant Mip-FlaA would be of great value in understanding the immunopathogenesis of the disease and in developing possible vaccine against the pathogen. Keywords Legionella pneumophila Epitope vaccine Mip FlaA Immunity
Introduction Legionella pneumophila, facultative parasitic bacteria, widely exist in natural and artificial water and soil. The bacteria can be inhaled by people in the form of aerosol and invade the macrophages of human body. In macrophages, the bacteria can effectively evade the damage effect of phagosome and lysosome, and their replication eventually leads to the death of the host cell [1]. The infection of L. pneumophila may cau
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