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Immunol Effects and Protect Efficiency Elicited by DNA Vaccine Contained Th1 Epitopes of Tuberculosis ESAT-6 Antigen and Flt-3 Ligand

Author: JiangQingTao
Tutor: WangYingWei
School: Nanjing Medical University
Course: Immunology
Keywords: Mycobacterium tuberculosis ESAT-6 FL DNA vaccines prime-boost
CLC: R392
Type: Master's thesis
Year: 2011
Downloads: 39
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Abstract


TB is a serious hazard to human health, infectious diseases, in recent years, TB worldwide epidemic. According to the latest report of the WHO, more than 1/3 of the world's population is infected with Mycobacterium tuberculosis (about 20 million) in HIV-infected patients, the incidence is even higher. China is the world's one of the 22 high TB ??burden countries, one of the countries with the most serious epidemic is also resistant TB epidemic situation is grim, arduous task of prevention. BCG (Mycobacterium bovis Bacillus Calmette-Guérin, BCG) is the only preventive vaccine Mycobacterium tuberculosis (Mycobacterium tuberculosis, M. tb) infection, better protection of its infant, but adults are ineffective, its protection efficiency between 0% -80%. Therefore, the development of new anti-tuberculosis vaccine has become an important issue to be solved by the medical profession. Our laboratory previously successfully constructed the Mycobacterium tuberculosis 6kD early secreted proteins (early secreted antigenic target ESAT-6) antigen Th1-type epitope fms-like tyrosine kinase 3 ligand (FL) recombinant DNA vaccine, this project aims to Therefore, the experiments focused on the following two aspects of the identification and evaluation of the vaccine effectiveness and protection efficiency. The first part of the identification of Mycobacterium tuberculosis ESAT-6 antigen Th1-type epitope FL recombinant DNA vaccine prime-boost strategy immune effects Objective: To study the Mycobacterium tuberculosis of ESAT-6 antigen the Th1 epitope FL recombinant plasmid (pIRES-TH-FL) on immune function. Method: plasmid pIRES-TH-FL mice immunized with a prime-boost strategy, check the mice specific lymphocyte proliferation; spleen cell culture supernatant and alveolar lavage fluid of Th1 and Th2 cytokines (IFN-γ, IL -12, IL-4, IL-10) secretion; mouse serum ESAT-6 specific IgG antibody levels; the frequency of IFN-γ T cells, as well as in vivo CTL killing function. Results: joint ESAT-6 protein Th1-type epitope with FL DNA vaccine can induce biased Th1-type immune response induced by prime-boost immunization programs better than BCG immunization effect. Conclusion: Mycobacterium tuberculosis ESAT-6 antigen Th1-type epitope FL recombinant DNA vaccine significantly increased the level of cellular immune responses in mice. The second part of the evaluation of Mycobacterium tuberculosis ESAT-6 antigen Th1-type epitope FL recombinant DNA vaccine prime-boost strategy protective effect Objective: To study the Mycobacterium tuberculosis of ESAT-6 antigen the Th1 epitope FL recombinant plasmid (pIRES-TH-FL) protection of mice against Mycobacterium tuberculosis. : Plasmid pIRES-TH-FL immune C57BL / 6 mice, and then attack Mycobacterium tuberculosis standard strain H37Rv, the protective effect of the vaccine from the colony counts and histological damage evaluation. Results: Th1-type epitopes of Mycobacterium tuberculosis ESAT-6 antigen FL recombinant DNA vaccine in mice, in the Netherlands the amount of bacteria in the mouse lung and spleen significantly reduce; HE staining and immunohistochemistry results showed that the degree of inflammatory infiltration significantly reduced . Prime-boost group compared with the separate DNA vaccine group, the protective effect is more obvious. Conclusion: ESAT-6 antigen Th1-type epitope FL recombinant DNA vaccine immunized mice that are resistant to the attack of Mycobacterium tuberculosis, provide effective protection, the prime-boost strategy to further enhance this protective effect.

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