左氧氟沙星联合CD25单克隆抗体治疗小鼠结核病的初步研究  

The effect of CD4^＋CD25^＋ regulatory T cell inactivation combined with levofloxacin on murine tuberculosis

作　　者：童新灯[1] 李美忠[2] 周伯平[2] 陈心春[2] 彭雁忠[1] 乐晓华[2] 苟继周[1] 唐志姣[3] 

机构地区：[1]武汉大学人民医院感染性疾病科,430060 [2]广东医学院附属深圳第三人民医院 [3]武汉大学医学院动物实验中心

出　　处：《中华结核和呼吸杂志》2009年第9期685-688,共4页Chinese Journal of Tuberculosis and Respiratory Diseases

基　　金：国家自然科学基金资助项目（30671857）

摘　　要：目的研究CD4^＋CD25^＋三调节性T细胞（Treg细胞）消除与左氧氟沙星联合治疗对小鼠结核病细胞免疫作用的影响。方法3～4周龄C57BL／6雌性小鼠76只，体重17～19g，随机数字表法分为对照组、CD25单克隆抗体（PC61）消除组、左氧氟沙星组和联合治疗组，每组19只。PC61消除组和联合治疗组小鼠腹腔注射含50μg PC61的PBS，0．2mE／只，对照组和左氧氟沙星组小鼠腹腔注射含50μg小鼠IgG同型对照的PBS，0．2mE／只。全部小鼠3d后尾静脉注射0．1ml的H37Rv（1×10。CFU）造模，左氧氟沙星组和联合治疗组小鼠于攻毒后第2天每只给予左氧氟沙星200mg·kg^-1·d^-1连续灌胃45d。分析小鼠体内不同组织中Treg细胞百分率及其对特异性细胞免疫、肺组织病理变化的影响。应用单因素方差分析对4组连续变量进行检验，组间比较采用q检验，组内不同时间点比较采用t检验。结果感染MTB第10天和第30天小鼠脾细胞中Treg细胞百分率：对照组分别为（30±4）％和（17．3±1．6）％，PC61消除组分别为（21±4）％和（16．1±1．3）％，左氧氟沙星组分别为（44±6）％和（24．7±2．0）％，联合治疗组分别为（24±3）％和（10．4±1．0）％，除第10天联合治疗组与PC61消除组比较无明显差别外，其他各组间比较均差异有统计学意义（q值为3．62～5．56，均P〈0．05）。联合治疗组小鼠的肺组织病变较轻，死亡数量较少。结论Treg细胞消除联合左氧氟沙星治疗可促进结核病小鼠的特异性细胞免疫，使肺内病变有所改善，并可降低小鼠的病死率。Objective To investigate the effects of inactivation of CD4 CD25＋ regulatory T cells （Treg） combined with the administration of levofloxacin（LFX） on the cellular immune response of murine tuberculosis. Methods Inactivation of Treg was achieved by intraperitoneal injection of anti-CD25, clone PC61. Female C57BL/6 mice were divided into 4 groups, PC61 alone, LFX alone, PC61 plus LFX, and the control, with 19 mice in each group. The LFX group and the control group were treated with rat-IgG isotope control. Mice were inoculated with ns7Rv （1× 10^6 CFU） via the tail vein 3 days later. From the 2nd day, the LFX group and the PC61 plus LFX group received intragastric administration of LFX at 200 mg· kg- 1 · d - 1 per mouse for 45 days. Blood and spleen Tregs were measured by flow cytometry. The cellular immune response and pulmonary histopathology at different time points were also evaluated after infection. Results At the 10th and 30th day, the ratio of CD4^＋ CD25/CD4 T cells in the spleen was （30 ± 4 ）% and （ 17. 3 ± 1.6） % respectively in the control group, （ 21 ± 4 ） % and （ 16. 1 ±1.3 ） % respectively in the PC61 group, （44±6）% and （24.7 ±2.0）% respectively in the LFX group, （24 ±3）% and （10.4 ± 1.0）% respectively in the PC61 plus LFX group. The differences were significant between groups （q = 3.62 -5.56, P 〈0. 05）, but the difference between the PC61 plus LFX group and the PC61 group at the lOth day. Same results were obtained from the peripheral blood. PC61 plus LFX therapy resulted in BCG specific cytokine response （IL-17, IFN-γ, TNF-α） from routine spleen cells at the 10th and the 30th day, and also in milder pathologic changes and the lowest mortality. Conclusions The cellular immune response was enhanced by Treg inactivation and LFX therapy, which decreased the pathologic changes and the mortality of murine tuberculosis.

关 键 词：T淋巴细胞 调节性 结核 肺 氧氟沙星 动物实验 

分 类 号：R686[医药卫生—骨科学]