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《Chinese Journal of Immunology》 2011-08
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Infection of macrophages with viable strain H37Ra and Heat-Killed H37Rv

HE Zong-Lin,DU Xian-Zhi.Department of Respiratory Medicine,the Second Affiliated Hospital,Chongqing Medical University,Chongqing 400010,China  
Objective:To study the difference in the expression of immune substances which macrophages are induced by inoculating mice with viable M.tuberculosis H37Ra and Heat-Killed M.tuberculosis H37Rv,and to compare the difference between viable and Heat-Killed M.tuberculosis in the induction of immune responses,and thus to learn whether viable or Heat-Killed bacteria could be a possible candidate for new vaccines.Methods:Peritoneal macrophages from BALB/c mouse were cultured with H37Ra and H37Rv for 10 hours respectively,to observe the situation of each group macrophages,and calculate the phagocytic rates of macrophages by the acid dye staining method.BALB/c mice were abdominally immunized with viable H37Ra,Heat-Killed H37Rv and NS,respectively.At 30 days after immunization,the macrophages were isolated.The expression levels of IL-12P40,TNF-α,IFN-γ were determined by RT-PCR and ELISA.The expression levels of NO,H2O2 were determined by Griess and Chemical method;and the changes of CD40L induced IFN-γ were detected by flow cytometry.Results:The phagocytic rates of macrophages were(55.71±8.42)% for viable M.tuberculosis and(14.82±2.12)% for Heat-Killed M.tuberculosis with a significant difference(P0.01).The mRNA levels of IL-12P40,TNF-α,IFN-γ were higher in viable M.tuberculosis H37Ra than those of in Heat-Killed M.tuberculosis,the cytokine levels of IL-12P40,TNF-α,IFN-γ,NO and H2O2 were higher in Viable M.tuberculosis H37Ra than those of in Heat-Killed M.tuberculosis.At the same time,the expressions of CD40L on macrophages induced by IFN-γ in viable M.tuberculosis stimulated group was higher than that of Heat-Killed M.tuberculosis stimulated group.Conclusion:Viable M.tuberculosis H37Ra can induce the activation of macrophages and the production of more protective immune substances,which play important roles in the host immune response,so it would be a candidate for tuberculosis vaccine.However,Heat-Killed M.tuberculosis H37Rv can't induce the activation of macrophages effectively and produce less protective immune substances,which makes it unlikely to be a candidate for tuberculosis vaccine.
【Fund】: 国家自然科学基金资助项目(No.30872261)
【CateGory Index】: R392.11
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