CAJ | 학술논문

目的研究CD_4~+CD_(25)~+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17～19 g,随机数字表法分为对照组、CD_(25)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50 μg PC61的PBS,0.2 ml/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,0.2ml/只.全部小鼠3 d后尾静脉注射0.1 ml的H_(37)Rv(1×10~6 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200 mg·kg~(-1)·d~(-1)连续灌胃45 d.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.
목적 연구CD_4~+CD_(25)~+조절성T세포(Treg세포)소제여좌양불사성연합치료대소서결핵병세포면역작용적영향.방법 3～4주령C57BL/6자성소서76지,체중17～19 g,수궤수자표법분위대조조、CD_(25)단극륭항체(PC61)소제조、좌양불사성조화연합치료조,매조19지.PC61소제조화연합치료조소서복강주사함50 μg PC61적PBS,0.2 ml/지,대조조화좌양불사성조소서복강주사함50μg소서IgG동형대조적PBS,0.2ml/지.전부소서3 d후미정맥주사0.1 ml적H_(37)Rv(1×10~6 CFU)조모,좌양불사성조화연합치료조소서우공독후제2천매지급여좌양불사성200 mg·kg~(-1)·d~(-1)련속관위45 d.분석소서체내불동조직중Treg세포백분솔급기대특이성세포면역、폐조직병리변화적영향.응용단인소방차분석대4조련속변량진행검험,조간비교채용q검험,조내불동시간점비교채용t검험.결과 감염MTB제10천화제30천소서비세포중Treg세포백분솔:대조조분별위(30±4)%화(17.3±1.6)%,PC61소제조분별위(21±4)%화(16.1±1.3)%,좌양불사성조분별위(44±6)%화(24.7±2.0)%,연합치료조분별위(24±3)%화(10.4±1.0)%,제제10천연합치료조여PC61소제조비교무명현차별외,기타각조간비교균차이유통계학의의(q치위3.62～5.56,균P<0.05).연합치료조소서적폐조직병변교경,사망수량교소.결론 Treg세포소제연합좌양불사성치료가촉진결핵병소서적특이성세포면역,사폐내병변유소개선,병가강저소서적병사솔.
Objective To investigate the effects of inactivation of CD_4~+ CD_25~+ 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-CD_(25), 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 H37Rv (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 CD_4~+ CD_(25)+/CD_4~+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 10th 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 murine 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.