乙肝患者HBV特异性细胞毒T淋巴细胞的分析  

Analysis of HBV-specific cytotoxic T lymphocytes in patients with hepatitis B

作　　者：丁宁[1] 黄丽利[1] 刘妍[1] 范振平[1] 刘立明[1] 赵平[1] 靳雪源[1] 叶海燕[1] 徐东平[1] 

机构地区：[1]解放军302医院传染病研究所与肝衰竭诊疗研究中心,北京100039

出　　处：《解放军医学杂志》2011年第4期361-364,共4页Medical Journal of Chinese People's Liberation Army

基　　金：国家自然科学基金(30771898);军队"十一五"医药卫生科研杰出人才项目(06J024);国家科技重大专项课题(2009ZX10005-017)

摘　　要：目的分析急性乙肝患者外周血中HBV特异性细胞毒T淋巴细胞(CTL)的表达特点及其在免疫应答机制中的作用。方法采集2008年10月-2010年6月于解放军302医院就诊的40例急性乙肝患者血样进行人白细胞抗原-A2(HLA-A2)检测,对其中18例HLA-A2阳性患者取病程极期及恢复期外周血,分离单个核细胞,并以6例HLA-A2阴性急性乙肝患者、18例HLA-A2阳性慢性乙肝患者和6例HLA-A2阳性健康正常人作为对照,经CD3、CD8单克隆抗体和HBV多肽抗原(HBsAg183-191,HB-sAg335-343)特异性五聚体(pentamer)染色后,采用流式细胞仪检测HBV特异性CTL频率,Cell-quest软件分析结果。同时检测患者发病极期丙氨酶转氨酶(ALT)峰值及HBV DNA载量,并进行相关性分析。结果 18例HLA-A2阳性急性乙肝患者急性期HB-sAg183-191的特异性CTL频率为0.23%±0.18%,HBsAg335-343特异性CTL频率为0.49%±0.31%,较HLA-A2阳性慢性乙肝患者发病极期上述抗原的特异性CTL频率(分别为0.07%±0.03%和0.08%±0.04%)明显升高(P<0.01);HBsAg183-191的特异性CTL频率与ALT峰值无显著相关性(r=0.4506,P=0.0528),而HBsAg335-343特异性CTL频率与ALT水平呈正相关(r=0.5642,P=0.0119),但上述CTL频率均与乙肝病毒载量无明显相关。与发病极期相比,患者在恢复期HBsAg335-343特异性CTL频率明显降低(P=0.011)。HLA-A2阴性急性乙肝患者及HLA-A2阳性正常人均未检出上述2种抗原的特异性CTL。结论 HBV特异性CTL在病毒清除和肝损伤中具有重要作用,针对不同HBV抗原表位的CTL,其作用可有一定差别。Objective To investigate the expression feature of hepatitis B virus（HBV）-specific cytotoxic T lymphocytes（CTL）,and analyze the role of CTL in immune response.Methods Peripheral venous blood was collected from 40 acute hepatitis B（AHB） patients hospitalized from Oct.2008 to Jun.2010 in 302 Hospital of PLA for the detection of human leucocyte antigen-A2（HLA-A2）.Out of the 40 patients,18 were HLA-A2-positive,and their peripheral blood mononuclear cells were isolated at acute phase and convalescent phase if the patients could be contacted.Six HLA-A2-negative AHB patients,18 HLA-A2-positive chronic HB patients and 6 HLA-A2-positive normal persons served as controls.The CTL was stained by fluorescence-labeled anti-CD3 monoclonal antibody,anti-CD8 monoclonal antibody and HBV epitopic pentamers（S183-191,S335-343）,and examined with flow cytometry,then analyzed by software Cell-quest.The peak value of alanine aminotransferase（ALT） and HBV load was detected at acute phase of patients,and the correlation was analyzed.Results The average frequencies were 0.23%±0.18% for HBsAg183-191-specific CTL and 0.49%±0.31% for HBsAg335-343-specific CTL at the acute phase of the 18 AHB patients,and they were significantly higher than those in patients with chronic HB,in whom CTL frequencies were 0.07%±0.03% for HBsAg183-191-specific CTL and 0.08%±0.04% for HBsAg335-343-specific CTL（P〈0.01）.There was no correlation between the frequency of HBsAg183-191-specific CTL and the peak value of ALT（r=0.4506,P=0.0528）,while the frequency of HBsAg335-343-specific CTL was correlated with ALT level（r=0.5642,P=0.0119）.Neither the two CTL frequencies showed correlation with virus load.Compared with the acute phase,the CTL frequencies obviously decreased at convalescent phase（P=0.011）.Both kinds of CTL were not detectable in HLA-A2-negative AHB patients and HLA-A2-positive healthy individuals.Conclusion HBV-specific CTL may play an important role in both virus clearance and liver impairment.CTL targeting to

关 键 词：肝炎病毒 乙型 HLA抗原 T淋巴细胞 细胞毒性 

分 类 号：R512.62[医药卫生—内科学]