中华传染病杂志
中華傳染病雜誌
중화전염병잡지
CHINESE JOURNAL OF INFECTIOUS DISEASES
2009年
12期
715-720
,共6页
童新灯%李美忠%周伯平%陈心春%彭雁忠%乐晓华%苟继周%唐志姣
童新燈%李美忠%週伯平%陳心春%彭雁忠%樂曉華%茍繼週%唐誌姣
동신등%리미충%주백평%진심춘%팽안충%악효화%구계주%당지교
CD4阳性T淋巴细胞%结核%肺%小鼠%免疫%细胞
CD4暘性T淋巴細胞%結覈%肺%小鼠%免疫%細胞
CD4양성T림파세포%결핵%폐%소서%면역%세포
CD4~+ CD25~+ regulatory T cell%Tuberculosis%pulmonary%Mice%Immunity%cellular
目的 研究CD4~+CD25~+调节性T淋巴细胞(Treg)在小鼠肺结核发生中的作用.方法 PC61处理组小鼠腹腔注射抗-CD25(PC61),50μg/只,消除体内的Treg,IgG对照组注射相同剂量的IgG同型对照抗体.3 d后静脉注射H37Rv 0.1 mL(含1×10~6CFU活菌),攻击小鼠.流式细胞技术分析PC61对小鼠体内不同组织中Treg的消除效果,体外分析Treg消除对特异性细胞免疫、肺组织病理学和肺组织内结核分枝杆菌生长情况的影响.采用方差齐性F检验和非配对t检验比较实验数据.结果 PC61处理组与IgG对照组小鼠脾细胞中Treg占CD4~+T淋巴细胞比例在第10天分别为(21.13± 3.58)%和(30.42± 4.20)%(f=2.38,P<0.05);第30天为(16.12± 1.26)%和(17.34± 1.62)%(t=0.84,P>0.05),Foxp3~+占CD4~+T淋巴细胞比例在第10天为(32.07± 3.95)%和(60.55± 5.48)%(t=5.96,P<0.05).外周血也有类似结果.在PC61处理组及IgG对照组,感染后第10、30和60天时体外培养小鼠脾细胞分泌的卡介苗特异性细胞因子1L-17(ng/L)分别为5.1± 0.9比0、43.1± 10.0比5.9±2.8、124.8± 5.8比102.5±8.1(t=7.90,t=5.10,t=3.19;均P<0.05);IFN-7(ng/L)分别为28.4±8.2比4.0± 1.3、685.9±128.6比418.7± 20.4、310.9± 119.7比32.8±7.5(t=4.21,t=8.43,t=3.27;均P<0.05);TNF-a(ng/L)分别为38.6±5.0比16.3±4.0、112.9± 12.3比71. 5± 12.6、86.2± 8.2比0(t=4.95,t=3.33,t=14.8;均P<0.05).PC61处理小鼠在感染早期(第10天)肺内结核菌量低于IgG对照小鼠(t=4.63.P<0.01),但后期比较差异无统计学意义.PC61处理小鼠感染后期(第120天)肺组织病理改变较IgG对照组为轻.结论 结核菌感染后小鼠Treg显著增加;Treg能有效抑制结核菌特异性细胞免疫.进而影响小鼠肺内结核菌的清除和结核病的发生.
目的 研究CD4~+CD25~+調節性T淋巴細胞(Treg)在小鼠肺結覈髮生中的作用.方法 PC61處理組小鼠腹腔註射抗-CD25(PC61),50μg/隻,消除體內的Treg,IgG對照組註射相同劑量的IgG同型對照抗體.3 d後靜脈註射H37Rv 0.1 mL(含1×10~6CFU活菌),攻擊小鼠.流式細胞技術分析PC61對小鼠體內不同組織中Treg的消除效果,體外分析Treg消除對特異性細胞免疫、肺組織病理學和肺組織內結覈分枝桿菌生長情況的影響.採用方差齊性F檢驗和非配對t檢驗比較實驗數據.結果 PC61處理組與IgG對照組小鼠脾細胞中Treg佔CD4~+T淋巴細胞比例在第10天分彆為(21.13± 3.58)%和(30.42± 4.20)%(f=2.38,P<0.05);第30天為(16.12± 1.26)%和(17.34± 1.62)%(t=0.84,P>0.05),Foxp3~+佔CD4~+T淋巴細胞比例在第10天為(32.07± 3.95)%和(60.55± 5.48)%(t=5.96,P<0.05).外週血也有類似結果.在PC61處理組及IgG對照組,感染後第10、30和60天時體外培養小鼠脾細胞分泌的卡介苗特異性細胞因子1L-17(ng/L)分彆為5.1± 0.9比0、43.1± 10.0比5.9±2.8、124.8± 5.8比102.5±8.1(t=7.90,t=5.10,t=3.19;均P<0.05);IFN-7(ng/L)分彆為28.4±8.2比4.0± 1.3、685.9±128.6比418.7± 20.4、310.9± 119.7比32.8±7.5(t=4.21,t=8.43,t=3.27;均P<0.05);TNF-a(ng/L)分彆為38.6±5.0比16.3±4.0、112.9± 12.3比71. 5± 12.6、86.2± 8.2比0(t=4.95,t=3.33,t=14.8;均P<0.05).PC61處理小鼠在感染早期(第10天)肺內結覈菌量低于IgG對照小鼠(t=4.63.P<0.01),但後期比較差異無統計學意義.PC61處理小鼠感染後期(第120天)肺組織病理改變較IgG對照組為輕.結論 結覈菌感染後小鼠Treg顯著增加;Treg能有效抑製結覈菌特異性細胞免疫.進而影響小鼠肺內結覈菌的清除和結覈病的髮生.
목적 연구CD4~+CD25~+조절성T림파세포(Treg)재소서폐결핵발생중적작용.방법 PC61처리조소서복강주사항-CD25(PC61),50μg/지,소제체내적Treg,IgG대조조주사상동제량적IgG동형대조항체.3 d후정맥주사H37Rv 0.1 mL(함1×10~6CFU활균),공격소서.류식세포기술분석PC61대소서체내불동조직중Treg적소제효과,체외분석Treg소제대특이성세포면역、폐조직병이학화폐조직내결핵분지간균생장정황적영향.채용방차제성F검험화비배대t검험비교실험수거.결과 PC61처리조여IgG대조조소서비세포중Treg점CD4~+T림파세포비례재제10천분별위(21.13± 3.58)%화(30.42± 4.20)%(f=2.38,P<0.05);제30천위(16.12± 1.26)%화(17.34± 1.62)%(t=0.84,P>0.05),Foxp3~+점CD4~+T림파세포비례재제10천위(32.07± 3.95)%화(60.55± 5.48)%(t=5.96,P<0.05).외주혈야유유사결과.재PC61처리조급IgG대조조,감염후제10、30화60천시체외배양소서비세포분비적잡개묘특이성세포인자1L-17(ng/L)분별위5.1± 0.9비0、43.1± 10.0비5.9±2.8、124.8± 5.8비102.5±8.1(t=7.90,t=5.10,t=3.19;균P<0.05);IFN-7(ng/L)분별위28.4±8.2비4.0± 1.3、685.9±128.6비418.7± 20.4、310.9± 119.7비32.8±7.5(t=4.21,t=8.43,t=3.27;균P<0.05);TNF-a(ng/L)분별위38.6±5.0비16.3±4.0、112.9± 12.3비71. 5± 12.6、86.2± 8.2비0(t=4.95,t=3.33,t=14.8;균P<0.05).PC61처리소서재감염조기(제10천)폐내결핵균량저우IgG대조소서(t=4.63.P<0.01),단후기비교차이무통계학의의.PC61처리소서감염후기(제120천)폐조직병리개변교IgG대조조위경.결론 결핵균감염후소서Treg현저증가;Treg능유효억제결핵균특이성세포면역.진이영향소서폐내결핵균적청제화결핵병적발생.
Objective To investigate the role of CD4 ~+ CD25~+ regulatory T lymphocytes (Treg)in modulating the cellular immune response and pathogenesis of murine pulmonary tuberculosis.Methods Inactivation of Treg was achieved by intraperitoneal injection anti-CD25 (clone PC61,50 μ/mouse) in PC61 group, and rat-IgG (50 μ/mouse) was injected intraperitoneally in control group. All the mice were inoculated intravenously with H37Rv 0. 1 mL (1 × 10~6 CFU) 3 days after Treg inactivation. The effects of Treg inactivation in different tissues were analyzed by flow cytometry. The cellular immune response, pulmonary histopathology and bacterial load were determined in vitro at different time points. The data were compared using homogeneity of variance F test and non-paired t test. Results In spleen, the percentages of Treg/CD4 T lymphocytes in PC61 group and control group were (21. 13± 3. 58)% and (30. 42± 4. 20)%, respectively at day 10 of inoculation (t = 2. 38, P < 0. 05), and those were (16. 12 ± 1. 26)% and ( 17. 34± 1. 62)%,respectively at day 30 of inoculation (t = 0. 84,P>0. 05). The percentages of Foxp3~+/CD4~+ T lymphocytes in PC61 group and control group were (32. 07 ± 3. 95)% and (60. 55 ± 5. 48)%,respectively at day 10 of inoculation (t = 5. 96, P<0. 05). Similar results were achieved in the peripheral blood. Bacillus calmette-guerin (BCG)-specific 1L-17 (ng/L) secreted by murine spleen cells in PC61 group and control group at day 10, 30 and 60 of inoculation were 5. 1± 0.9 vs 0, 43. 1± 10.0 vs5. 9± 2. 8 and 124.8 ± 5.8 vs 102. 5±8. 1, respectively (t = 7. 90, t=5. 10,t = 3. 19; all P<0.05); those of BCG-specific IFN-γ (ng/L) were 28. 4 ± 8. 2 vs 4. 0±1. 3, 685. 9± 128. 6 vs418. 7±20.4 and 310.9±119. 7 vs 32. 8±7. 5, respectively(tO = 4. 21,t = 8. 43, t = 3. 27; all P<0.05);those of TNF-a (ng/L) were 38. 6±5.0 vs 16. 3±4. 0, 112. 9 ±12. 3 vs 71. 5±12. 6 and 86. 2±8. 2vs0, respectively(t = 4. 95, t=3. 33,t/=14.8; all P<0. 05). The lung bacterial load at day 10 of inoculation in PC61 group was lower than that in control group (t = 4. 63, P < 0. 01), but the differences were not significant thereafter. The changes of lung histopathology at late stage of infection (day 120) in PC61 group were less severe than those in control group. Conclusions Murine Tregs increase dramatically after Mycobacterium tuberculosis infection. Treg could inhibit the specific cellular immunity against Mycobacterium tuberculosis, and therefore, may facilitate the persistent infection of Mycobacterium tuberculosis and development of tuberculosis.
