重组耻垢分枝杆菌对小鼠结核病免疫治疗效果的评价  

Therapeutic effects of GLS/IL-12 recombinant M. smegmatis in murine model of M. tuberculosis infection

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

机构地区：[1]重庆医科大学免疫学与微生物学教研室,重庆400016

出　　处：《免疫学杂志》2007年第4期357-361,共5页Immunological Journal

基　　金：国家自然科学基金资助项目(30400375)

摘　　要：目的评估携带编码人天然颗粒溶素(GLS)和小鼠IL-12基因质粒(pZM03)的重组耻垢分枝杆菌对结核分枝杆菌感染的治疗效果,及其与临床抗痨药物疗效的比较。方法结核分枝杆菌H37Rv感染Balb/c小鼠4周后分别用生理盐水、GLS/IL-12重组耻垢分枝杆菌(重组M.S)I、NH+PZA和重组M.S+INH+PZA治疗,于第1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分泌水平、肺泡灌洗液SIgA的水平和肺、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M.SI、NH+PZA和重组M.S+INH+PZA治疗组肺、脾组织荷菌量(logCFU/g)比生理盐水组显著降低。重组M.S组经PPD刺激后IFN-γ和TNF-α分泌水平明显高于其它组。重组M.S组血清IL-12和IFN-γ水平明显高于其它组。各组产生黏膜特异性SIgA明显高于未经感染组。用免疫组化检测到小鼠肺及脾组织中GLS的表达。生理盐水组肺组织病理改变以渗出为主,病变广泛,正常肺泡结构被破坏;其余组肺组织病变轻且局限。结论重组M.S对小鼠结核病有一定免疫治疗作用,通过增强宿主Th1型免疫应答和GLS的抗菌活性有关;重组M.S对临床常用的抗结核药有协同作用,为结核病的综合治疗打下实验基础。Objective To evaluate the effects of recombinant M. smegmatis （M. S） encoding human granulysin （GLS） and murine IL-12 on murine M. tuberculosis infection.Methods Balb/c mice infected with M. tuberculosis were treated with normal saline, recombinant M.S, INH （lsoniazid） ＋ PZA （pyrazinamide）, recombinant M.S ＋ INH＋ PZA, respectively. The numbers of viable bacteria in the lungs and the spleens were counted. ELISA was used to detect the levels of ILl2 and IFN-γ in serum, the SIgA level in bwnchoalveolar lavage fluid （BALF）, and the levels of IFN-γ and TNF-α released from PPD-stimulated spleen lymphocytes. The expression of GLS in tissue was detected by immunohistochemistry. Lungs and spleens were prepared for pathological analysis.Re,fits The recombinant M.S, INH ＋ PZA, and recombinant M.S ＋ INH ＋ PZA groups showed significantly reduce in number of colony forming units （log CFU/g） compared with saline group. The expression of GLS in tissue, the increase of serum IL-12, serum IFN-γ and BALF SIgA were found in recombinant M.S group. IFN-γ and TNF-α released from PPD-stimulated spleen lymphocytes in recombinant M.S group were higher than that of other groups. The pathological changes in lungs of the recombinant M. S group, INH ＋ PZA group, and recombinant M. S ＋ INH ＋ PZA group were localized, while that of the control group was extensive. Conclusion The recombinant M. S has immunotherapeutic effects on murine M. tuberculosis infection, which are associated with Thl response and antibacterial activity of GLS. The recombinant M. S has synergistic effects on anti-tuberculosis chemotherapy.

关 键 词：GLS IL-12 重组耻垢分枝杆菌 结核病 

分 类 号：R392[医药卫生—免疫学]