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Issue:ISSN 1000-7083
          CN 51-1193/Q
Director:Sichuan Association for Science and Technology
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Your Position :Home->Past Journals Catalog->2013 Vol.32 No.4

Immune Response of BALB/C Mice Induced by Ipr1/PPE68 Recombinant BCG
Author of the article:ZHANG Zhuangmiao1,2, YANG Chun1,2*, XU Lei1,2, HE Yonglin1,2, WANG Jingxian1,2, DONG Zhiling1,2, YAN
Author's Workplace:(1. Department of Pathogen Biology, Chongqing Medical University, Chongqing 400016, China; 2. Institute of Molecular Medicine and Cancer, Chongqing Medical University, Chongqing 400016, China)
Key Words:Mycobacterium tuberculosis (MTB); Ipr1/PPE68-rBCG; immune response
Abstract:Objective  To construct the recombinant BCG with intracellular pathogen resistance 1 (Ipr1) gene and coding region of Pentose-5-phosphate-3-epimerase 68 (PPE68), and to study the relevant immune response of rBCG immunized BALB/C mice. Methods  Ipr1 and PPE68 genes and OriM were cloned into the MCS sites of plasmid pBudCE4.1, respectively. Then the recombinant plasmid pBIPO was verified by enzyme restriction, DNA sequencing and Western-blotting. Subsequently, pBIPO was electro-transferred into BCG and positive colonies were analyzed by PCR. BALB/C mice were then immunized with Ipr1/PPE68-rBCG for two weeks, and the productions of IgG2a, IL-12, IFN-γ and IL-4 in the sera were detected by ELISA, the quantity of CD4+ and CD8+ T cells were assessed by FACS, specific spleen lymphocytes proliferations were evaluated by MTT. At the mean time, numbers of residual Mycobacterium tuberculosis and pathological changes in different organs of BALB/C mice were investigated. Results  Enzyme restriction, DNA sequencing and Western-blotting confirmed that the Ipr1/PPE68-rBCG was successfully constructed. Compared with control group, Ipr1/PPE68-rBCG immunized mice showed a significantly increased production of IgG2a and IL-12 in the sera, and enhanced spleen lymphocyte proliferative responses. However, no significant difference was observed between Ipr1/PPE68-rBCG group and BCG group. Although the productions of IFN-γ in the sera were significantly lower than BCG group, no significant difference was observed between Ipr1/PPE68-rBCG group and control group. All the group showed an equal production of IL-4. No residual Mycobacterium tuberculosis was found, and there were no obvious pathological changes in the lung and spleen. Conclusion  Ipr1/PPE68-rBCG were successfully constructed which could induce effective cellular immunity in BALB/C mice.
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