结核分枝杆菌Mtb8.4基因疫苗与人白细胞介素12表达质粒联合免疫的细胞免疫应答观察  

Observation on cellular immune response in mice induced by co-immunization of Mtb8.4 gene vaccine and plasmid encoding human interleukin 12

作　　者：李晖[1] 李强[1] 钟森[2] 任红[3] 邓存良[1] 

机构地区：[1]泸州泸州医学院附属医院感染病科,四川646000 [2]成都中医药大学附属医院 [3]重庆医科大学第二临床学院感染科

出　　处：《解放军医学杂志》2007年第3期229-230,244,共3页Medical Journal of Chinese People's Liberation Army

基　　金：四川省青年科技基金资助项目(川青科基[2002]1);四川省重点学科重点建设项目(SZD0241)

摘　　要：目的观察结核分枝杆菌Mtb8.4基因疫苗与人白细胞介素12(hIL-12)联合免疫小鼠所诱导的细胞免疫应答。方法15只C57BL/6N小鼠随机分为Mtb8.4基因疫苗+hIL-12质粒组(联合免疫组)、Mtb8.4基因疫苗组、卡介苗(BCG)组、空载体组和PBS组,基因疫苗、空载体和PBS经肌内注射法免疫各组小鼠,每隔3周免疫1次,共免疫3次,BCG组经尾部皮下注射1×106CFUBCG免疫1次。ELISA法检测小鼠脾细胞培养上清中细胞因子水平;乳酸脱氢酶(LDH)释放法检测免疫小鼠特异性细胞毒性T淋巴细胞(CTL)杀伤活性。结果联合免疫组能诱导较强的抗原特异性Th1型细胞免疫应答,免疫小鼠脾细胞培养上清液IFN-γ和IL-2水平(分别为1493.340±8.128pg/ml、747.489±48.676pg/ml)显著高于Mtb8.4基因疫苗组,与BCG组相当,IL-4分泌减少,特异性CTL杀伤活性增强。结论hIL-12表达质粒能够增强Mtb8.4基因疫苗所诱导的细胞免疫应答。Objective To observe the specific cellular immune response induced by co-immunization of DNA vaccine of Mtb8. 4 and plasmid encoding human interleukin 12 （hIL-12） in mice. Methods Fifteen C57BL/6N mice were divided into following groups： Mtb8. 4 gene vaccine plus plasmid of hIL-12, Mtb8.4 gene vaccine, BCG, empty vector alone and PBS. Mice were immunized intramuscularly in both hind limbs three times at the intervals of three weeks or once subcutaneously with 1 × 10^6 of viable M. bovis BCG Pasteur at the time of the first DNA immunization. The level of IFN-γ in supernatant of spleno-lymphocyte cultures was measured by ELISA. CTL activities of spleno-lymphocyte were detected with LDH release assay. Results The levels of IFN-γ and IL-2 in supernatant of spleno-lymphocyte cultures in the group of Mtb8. 4 gene vaccine plus plasmid of hIL-12 were significantly higher than that of group of Mtb8. 4 alone （P〈 0. 01）, and they were similar to that in BCG group, the level of IL-4 decreased and was lower than that of BCG group （P〈0. 01）. CTL activity of spleno-lymphocyte in the group of Mtb8.4 gene vaccine plus plasmid of hIL-12 was stronger than that in other groups. Conclusion Co-immunization of Mtb8. 4 gene vaccine and plasmid encoding hIL-12 can enhance the specific cellular immune response induced by the Mtb8. 4 gene vaccine alone.

关 键 词：分枝杆菌 结核 基因 MTB8.4 白细胞介素12 联合免疫 

分 类 号：R378.911[医药卫生—病原生物学]