中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2009年
5期
411-415
,共5页
王孝勤%张野%白雪帆%黄长形%连建奇
王孝勤%張野%白雪帆%黃長形%連建奇
왕효근%장야%백설범%황장형%련건기
乙型肝炎肝硬化%Toll样受体%CD4+CD25+调节性T细胞%流式细胞术
乙型肝炎肝硬化%Toll樣受體%CD4+CD25+調節性T細胞%流式細胞術
을형간염간경화%Toll양수체%CD4+CD25+조절성T세포%류식세포술
HBV-related liver cirrhosis%Toll-like receptor%CD4+ CD25 + regulatory T cells%Flow cytometry
目的 检测乙型肝炎肝硬化患者外周血单个核细胞(peripheral blood mononuelear cells,PBMCs)表面Toll样受体(Toll-like receptor,TLR)2和TLR4以及CD4+CD25+调节性T细胞(regulato-ry T cells,Treg)的表达,并探讨TLR2、TLR4与Treg的相关性.方法 分别应用抗-TLR2-PE、抗-TLR4-APC、抗-CD14-FITC及抗-CD4-PerCP、抗-CD25-FITC、扰-CD127-PE对67例研究对象[乙型肝炎肝硬化患者(HBV related liver cirrhosis,LC)30例、慢性乙型肝炎患者(chronic hepatitis B,CHB)21例、健康对照(normal control,Nc)16例]PBMCs表面分子进行免疫荧光染色,应用流式细胞仪分别对TLR2、TLR4及Treg进行检测.组间比较采用Kruskal-wallis秩和检验,相关性分析采用Spearman秩相关.结果 LC组、CHB组、NC组单核细胞表面TLR2、TLR4的平均荧光强度(MFI)分别为200.3±96.8和32.1±7.2、214.0±72.6和25.2 ±8.3、94.1 ±17.6和17.8 ±3.9.LC组与NC组、CHB组与NC组TLR2 MFI比较差异均有统计学意义(P<0.05),LC组与CHB组TLR2 MFI差异无统计学意义(P>0.05).LC组与NC组、CHB组与NC、LC与NC组TLR4 MFI比较差异均有统计学意义(P<0.05).LC组、CHB组、Nc组Treg占CD4+T淋巴细胞的比例分别为(5.07%±1.43%、5.88%±1.66%、4.21%±1.24%),CHB组与NC组、CHB组与LC组比较差异有统计学意义(P<0.05),而LC组与NC组比较差异无统计学意义(P>0.05).在LC组,TLR4的表达与Treg呈正相关(r=0.469,P=0.032),TLR2与血清病毒载量呈负相关(r=-0.428,P=0.021).结论 LC患者,TLR2、TLR4的表达显著上调,TLR2、TLR4可能与乙型肝炎肝硬化的发生有关.
目的 檢測乙型肝炎肝硬化患者外週血單箇覈細胞(peripheral blood mononuelear cells,PBMCs)錶麵Toll樣受體(Toll-like receptor,TLR)2和TLR4以及CD4+CD25+調節性T細胞(regulato-ry T cells,Treg)的錶達,併探討TLR2、TLR4與Treg的相關性.方法 分彆應用抗-TLR2-PE、抗-TLR4-APC、抗-CD14-FITC及抗-CD4-PerCP、抗-CD25-FITC、擾-CD127-PE對67例研究對象[乙型肝炎肝硬化患者(HBV related liver cirrhosis,LC)30例、慢性乙型肝炎患者(chronic hepatitis B,CHB)21例、健康對照(normal control,Nc)16例]PBMCs錶麵分子進行免疫熒光染色,應用流式細胞儀分彆對TLR2、TLR4及Treg進行檢測.組間比較採用Kruskal-wallis秩和檢驗,相關性分析採用Spearman秩相關.結果 LC組、CHB組、NC組單覈細胞錶麵TLR2、TLR4的平均熒光彊度(MFI)分彆為200.3±96.8和32.1±7.2、214.0±72.6和25.2 ±8.3、94.1 ±17.6和17.8 ±3.9.LC組與NC組、CHB組與NC組TLR2 MFI比較差異均有統計學意義(P<0.05),LC組與CHB組TLR2 MFI差異無統計學意義(P>0.05).LC組與NC組、CHB組與NC、LC與NC組TLR4 MFI比較差異均有統計學意義(P<0.05).LC組、CHB組、Nc組Treg佔CD4+T淋巴細胞的比例分彆為(5.07%±1.43%、5.88%±1.66%、4.21%±1.24%),CHB組與NC組、CHB組與LC組比較差異有統計學意義(P<0.05),而LC組與NC組比較差異無統計學意義(P>0.05).在LC組,TLR4的錶達與Treg呈正相關(r=0.469,P=0.032),TLR2與血清病毒載量呈負相關(r=-0.428,P=0.021).結論 LC患者,TLR2、TLR4的錶達顯著上調,TLR2、TLR4可能與乙型肝炎肝硬化的髮生有關.
목적 검측을형간염간경화환자외주혈단개핵세포(peripheral blood mononuelear cells,PBMCs)표면Toll양수체(Toll-like receptor,TLR)2화TLR4이급CD4+CD25+조절성T세포(regulato-ry T cells,Treg)적표체,병탐토TLR2、TLR4여Treg적상관성.방법 분별응용항-TLR2-PE、항-TLR4-APC、항-CD14-FITC급항-CD4-PerCP、항-CD25-FITC、우-CD127-PE대67례연구대상[을형간염간경화환자(HBV related liver cirrhosis,LC)30례、만성을형간염환자(chronic hepatitis B,CHB)21례、건강대조(normal control,Nc)16례]PBMCs표면분자진행면역형광염색,응용류식세포의분별대TLR2、TLR4급Treg진행검측.조간비교채용Kruskal-wallis질화검험,상관성분석채용Spearman질상관.결과 LC조、CHB조、NC조단핵세포표면TLR2、TLR4적평균형광강도(MFI)분별위200.3±96.8화32.1±7.2、214.0±72.6화25.2 ±8.3、94.1 ±17.6화17.8 ±3.9.LC조여NC조、CHB조여NC조TLR2 MFI비교차이균유통계학의의(P<0.05),LC조여CHB조TLR2 MFI차이무통계학의의(P>0.05).LC조여NC조、CHB조여NC、LC여NC조TLR4 MFI비교차이균유통계학의의(P<0.05).LC조、CHB조、Nc조Treg점CD4+T림파세포적비례분별위(5.07%±1.43%、5.88%±1.66%、4.21%±1.24%),CHB조여NC조、CHB조여LC조비교차이유통계학의의(P<0.05),이LC조여NC조비교차이무통계학의의(P>0.05).재LC조,TLR4적표체여Treg정정상관(r=0.469,P=0.032),TLR2여혈청병독재량정부상관(r=-0.428,P=0.021).결론 LC환자,TLR2、TLR4적표체현저상조,TLR2、TLR4가능여을형간염간경화적발생유관.
Objective To detect circulating CD4 + CD25 + regulatory T cells (Treg) and Toll-like receptor(TLR)2 and TLR4 expression on the peripheral blood mononuclear cells (PBMCs) of patients with HBV-related liver cirrhosis (LC), and to explore the correlation between them. Methods PBMCs isolated from 30 LC patients, 21 chronic hepatitis B (CHB) patients and 16 normal controls(NC) were stained with fluorescent labeling anti-TLR2-PE, anti-TLR4-APC, anti-CD14-FITC monoclonal antibodies and anti-CD4-PerCP, anti-CD25-FITC, anti-CD127-PE. Samples were detected by flow cytometry. Statistic analysis be-tween groups was performed by Kruskal-Wallis H test. Spearman rank correlation was used to analyze the correlation of Treg and TLR2, TLR4. Results The expression of TLR2 and TLR4 were significantly up-reg-ulated in patients with LC than those in the controls (TLR2 : 200.3 ± 96.8 vs 94.1 ± 17.6, P < 0.05 ; TLR4:32.1 ±7.2 vs 17.8 ±3.9, P<0.05). The expression of TLR4 was significantly increased in pa-tients with LC than those in patients with CHB (TLR4 : 32. 1 ± 7.2 vs 25.2 ± 8.3, P < 0.05), but there were no differences of TLR2 expression between LC and CHB(200.3 ± 96.8 vs 214.0 ± 72.6, P > 0.05). Treg/CD4+ T cells were 5.07% ±1.43%, 5.88% ±1.66%, 4.21% ±1.24% in patients with LC, CHB and NC, respectively. Treg/CD4+ T cells were significantly increased in patients with CHB than those in pa-tients with NC(P<0. 05) and LC(P <0.05), but there were no differences between LC and NC(P > 0.05). TLR4 expression and Treg were positive correlation (r = 0. 469, P = 0. 032) and TLB2 expression were negative correlation in patients with LC (r = -0.428, P = 0.021). Conclusion The expression of TLR2 and TLR4 were up-regulated on PBMCs in patients with LC. It seems to be expression of TLR2 and TLR4 in-volved in the pathogenesis of LC.
