重组BCG联合抗痨药物对小鼠结核病治疗作用的实验研究  

Evaluation synergism of GLS/IL-12 recombinant BCG to anti-tuberculosis chemotherapy in murine model of M.tuberculosis infection

作　　者：杨春[1] 张黎[1] 何永林[1] 徐蕾[1] 伊正君[1] 李娜[1] 王渝伟[1] 朱道银[1] 

机构地区：[1]重庆医科大学免疫学和微生物学教研室,重庆400016

出　　处：《中国免疫学杂志》2008年第5期452-455,460,共5页Chinese Journal of Immunology

基　　金：国家自然科学基金资助项目(No.30400375;307719227)

摘　　要：目的:探讨携带编码人天然颗粒溶素(Granulysin,GLS)和小鼠IL-12基因的重组BCG与抗痨药物对结核分枝杆菌感染的协同治疗效果。方法:将结核分枝杆菌H37Rv感染的BALB/c小鼠40只随机分成4组。感染4周后分别用生理盐水、重组卡介苗(BCG)、INH+PZA和重组BCG联合INH+PZA治疗,于第一次治疗后3个月,处死小鼠,检测器官荷菌量、脾淋巴细胞IFN-γ和TNF-α的分泌水平、血清IL-12和IFN-γ、肺、脾组织中GLS表达,同时观察小鼠肺、脾组织病理改变情况。结果:重组BCG联合INH+PZA治疗组肺、脾组织荷菌量(logCFU/g)比重组BCG和INH+PZA治疗组显著降低;重组BCG和INH+PZA治疗组肺、脾组织荷菌量比生理盐水组显著降低。重组BCG组经PPD刺激后IFN-γ和TNF-α分泌水平明显高于其它组。重组BCG联合INH+PZA治疗组和重组BCG组血清IL-12和IFN-γ水平明显高于生理盐水组。重组BCG组用免疫组化检测到小鼠肺及脾组织中GLS的表达。重组BCG和INH+PZA组病变轻且局限,生理盐水组肺组织病理改变以渗出为主,病变广泛。重组BCG联合INH+PZA治疗组肺组织病变最轻。结论:GLS/IL-12重组BCG对小鼠结核病有一定免疫治疗作用;重组BCG联合临床常用的抗痨药可增强抗结核病的疗效。Objective： To evaluate the synergism of recombinant Bacillus Calmette-Guerin （BCG） encoding human granulysin and murine IL-12 to anti-tuberculosis chemotherapy in murine M.tuberculosis infection.Methods： BALB/c mice were infected with M. tuberculosis and were injected with normal saline,recombinant BCG, INH（Isoniazid） ＋ PZA（pyrazinamide）, recombinant BCG ＋ INH ＋ PZA respectively.The numbers of viable bacteria in lungs and spleens were cotmted.The levels of IL-12 and IFN-γ in serum and IFN-γ and TNF-α released from spleen lymphocytes stimulated with PPD were detected with ELISA. The expression of GLS in tissue was detected with immtmohistochemistry. Lungs and spleens were prepared for pathological analysis. Results：The recombinant BCG group（4. 248 ± 0.414 in lung, 3.538 ± 0.563 in spleen）, INH ＋ PZA group（3.702 ± 0.438 in 1 ,ung, 3. 320 ± 0. 358 in spleen）, and recombinant BCG ＋ INH ＋ PZA group （3.258 ± 0.250 in lung, 2.011 ± 0.103 in spleen）showed significantly reduced number of colony forming unlts（log CFU/g）compared with saline group. The expression of GLS in tissue and the increased of IL-12 and IFN-γ in serum were found in recombinant BCG group. IFN-γ and TNF-α released from spleen lymphocytes stimulated with PPD in recombinant BCG group were higher than that of other groups（ P 〈 0.01）. The pathological changes in lungs of the recombinant BCG group,INH＋ PZA group,and recombinant BCG ＋ INH＋ PZA group were localized,while was spread in the control group. The significant pathological changes were not found in the spleens of all groups. Condusion： The recombinant BCG has immtmotherapeutic effects,the recombinant BCG has a synergism to anti-tuberculosis chemotherapy.

关 键 词：颗粒溶素 IL-12 重组BCG 结核病 抗痨药 

分 类 号：R378.911[医药卫生—病原生物学]