人结核分枝杆菌hsp70 DNA疫苗的免疫保护作用研究  

A study on the protective immune function of Hsp70 DNA Vaccine in Human Mycobacterium Tuberculosis

作　　者：胡佳杰[1] 文学明[1] 肖爱清[2] 黄海浪[3] 

机构地区：[1]湖北药检高等专科学校检验系,430064 [2]湖北省疾病控制中心 [3]华中科技大学同济医学院分子生物学教研室

出　　处：《贵州医药》2002年第10期867-869,共3页Guizhou Medical Journal

基　　金：国家自然科学基金 (NO:394 80 0 2 2 )资助项目

摘　　要：目的研究人结核分枝杆菌hsp70DNA疫苗对小鼠的免疫保护作用。方法用人结核分枝杆菌hsp70DNA疫苗免疫雄性BALB c小鼠 ,8周后 ,用结核杆菌H37Ra进行攻击 8周 ,检测小鼠淋巴细胞刺激指数 (SI)、腹腔巨噬细胞培养上清释放NO量、小鼠血清IFN γ、IL 2的含量 ,并计数小鼠肝、肺活细菌数。结果hsp70DNA疫苗免疫的小鼠 ,其脾淋巴细胞刺激指数 (SI)为 3 2 4± 0 57,与对照组 (2 63± 0 47)比较 ,差异不具有显著意义 ;腹腔巨噬细胞培养释放NO量与对照组比较无显著差异 ,但较对照组高 2 6 %。免疫小鼠其血清IFN γ含量为 981 40pg ml,与对照组 (565 90pg ml)相比 ,差异具有显著意义 (P <0 0 5) ;小鼠血清中IL 2的浓度较对照组明显提高 (P <0 0 5)。经hsp70DNA疫苗免疫的小鼠受结核杆菌攻击后其肺、肝脏结核杆菌数较对照组少。结论人结核分枝杆菌hsp70DNA疫苗免疫小鼠后能提高小鼠腹腔巨噬细胞培养上清释放NO量 ,刺激IFN γ、IL 2的分泌 ,说明此疫苗能诱导CD+4 Th1和CD+8CTL的细胞免疫反应 ,增强小鼠细胞免疫功能 。objective To study the protective immune function of hsp70 DNA vaccine in human Mycobacterium tuberculosis. Methods BALB/c male mice were immunized with hsp70 DNA vaccine in human Mycobacterium tuberculosis. Eight weeks after immunization, the mices were subsequently challenged with M.tuberculosis H37Ra for eight weeks. Stimulating index(SI) of the evaluated splenic lymphocyte transformation and the amounts of NO in supernatant released from intraperitoneal macrophages cultured were detected. The levels of IFN γand IL 2 in the serum were detected.Bacterial CFU in livers and lungs were calculated. Results The SI of the experimental group was 3.24±0.57, which was significantly increased as compared with that in the vector group (1.56±0.06) and there was no statistical difference compared with the control group (2.63±0.47, P >0.05). There was no statistical difference about the amounts of NO in supernatant released from the intraperitoneal macrophages of the mice between all groups, but the amounts of NO of the experimental group increased by 26%. The serum IFN γ level of the experimental group was 981.40pg/ml±332.10pg/ml, which was significantly higher than those in the control group( P <0.05). The serum IL 2 level of the experimental group was significantly higher than in the control group. Live bacterial CFU of livers were lower than those in the control group. Conclusion The vaccine could effectively stimulate host cells to generate IL 2 and IFN γ. The vaccine could elicit antigen specific CD + 8 CTL and CD + 4 Th1 responses and induce extremely strong protective immunity against subsequent challenge with M.tuberculosis H37Ra.

关 键 词：结核分枝杆菌 hsp70DNA疫苗 免疫保护 动物实验 

分 类 号：R979.5[医药卫生—药品] R52[医药卫生—药学]