血清sICAM1测定在判定乙型肝炎不同临床类型中的意义
(西安交通大学：第二医院检验科，陕西 西安 710004 ，医学院免疫学教研室，陕西 西安 710061)
Significance of serum sICAM1 in the clinical types of hepatitis B
WANG XiangLing, WANG XiaoLi, ZHAO LiHua, LI MiaoXian, ZHANG XiYing， XIE Ming, JI YuQiang
Department of Clinical Laboratory, Second Hospital, Xian Jiaotong University，Xian 710004，China，Department of Immunology，Medical College, Xian Jiaotong University，Xian 710061, China
【Abstract】 AIM: To observe the changes and significance of serum sICAM1 in the assessing clinical types of Hepatitis B. METHODS: Double antibody sandwich ABCELISA was employed for the quantitative measurement of serum sICAM1 of 74 cases of different clinical types and 10 cases of normal control . RESULTS: Both the sICAM1 level of 13 cases of acute hepatitis B (680±248) μg·L-1 and that of 46 cases of chronic hepatitis B (675±311) μg·L-1 were significantly higher than that of normal control (204±45) μg·L-1. The sICAM1 level of 18 cases of moderate chronic hepatitis B (634±182) μg·L-1 was significantly higher than the 19 cases of mild chronic hepatitis B (445±120) μg·L-1. The sICAM1 level of 9 cases of serious chronic hepatitis B (931±280) μg·L-1 was significantly higher than that of the group of mild chronic hepatitis B. The sICAM1 level of 10 cases of completely recovered cases (243±86) μg·L-1 and that of 5 cases of asymptomtic carriers (212±72) μg·L-1 were not significantly different from that of the normal control. Serum concentrations of sICAM1 were significantly positively correlated with the concentrations of ALT. There was no significant difference between the HBeAg positive group and HBeAg negative group. CONCLUSION: Detection of sICAM1 can be used as a indicator in assessing clinical types. The concentration of sICAM1 is positivley correlated with the level of liver damage but is not related to the virus duplication.
【Keywords】 sICAM1; Hepatitis B; clinical types
【摘 要】目的: 观察血清sICAM-1水平在HBV感染者不同临床类型中的变化及其意义.方法 ： 采用双抗体夹心ABC-ELISA法，对74例不同临床类型HBV感染者及10例正常对照者血清中可溶性细胞间粘附分子-1(sICAM1)进行了定量测定. 结果 ： 急性肝炎13例血清中sICAM-1含量(680±248) μg·L-1明显高于正常对照组(204±46) μg·L-1 (P<001); 46例慢性肝炎患者血清中sICAM-1含量(675±311) μg·L-1明显高于正常对照组(P<001);其中慢性肝炎轻度19例，sICAM-1含量(445±120) μg·L-1；慢性肝炎中度18例，sICAM-1含量为(634±182)μg·L-1，明显高于慢性轻度组(P<001)；慢性肝炎重度9例，sICAM1含量(931±280)μg·L-1，明显高于慢性轻度组(P<001).5例无症状携带者sICAM1含量(212±72) μg·L-1，与正常对照组差别无显著性(P>005); 10例自然恢复组含量为(243±86) μg·L-1，与正常对照组差别无显著性.血清中sICAM1含量与ALT含量呈明显正相关;而E抗原阳性组与E抗原阴性组差异无显著性.结论： 血清中sICAM-1水平测定可作为判别乙型肝炎临床类型的指标；sICAM1含量与肝脏损伤程度成正比，sICAM1含量与病毒复制状态无关.
【关键词】 sICAM1；肝炎乙型，临床类型
0 引言
乙型肝炎为全球性传染病,根据病程可分为急性乙型肝炎, 慢性乙型肝炎.根据临床表现可分为轻型、中型、重型乙型肝炎和无症状乙肝病毒携带者.可发展为肝硬化、肝癌.目前尚无确切可靠的特异治疗方法,也无提示其预后的明确指标.近年来大量研究认为, 乙型肝炎不同的临床类型与机体抗病毒免疫应答密切相关.ICAM1能介导免疫反应中各类细胞间的粘附，目前研究认为，ICAM1/LFA1介导的细胞粘附在炎症性肝病时T细胞与肝细胞相互作用中起着关键性作用.为观察乙型肝炎不同临床表现者细胞间黏附分子的变化,我们测定了74例不同临床类型乙肝感染者血清中sICAM1水平, 报告如下.
1 对象和方法
1.1 对象 乙肝患者59(男45，女14)例，年龄7~51岁，其中急性肝炎13例，慢性肝炎轻度19例，中度18例，重度9例，均为我院传染科住院或门诊患者，用免疫制剂治疗前采集标本；无症状携带者5(男4，女1)例，年龄16~46岁，为门诊复检者，检出HBsAg阳性超过1a；自然恢复者10(男6，女4)例，年龄15~52岁，为门诊复检者，HBVM检测为抗HBc和抗HBs阳性或抗HBe，抗HBc，抗HBs阳性或抗HBs阳性，未注射乙肝疫苗；正常对照10(男7，女3)例，年龄15~55岁，均为门诊健康体检者，未注射乙肝疫苗.常规静脉采血，4 h内分离血清，检测乙肝病毒标志物及ALT，同时分出血清于-30℃保存.sICAM1试剂购于上海森雄公司；HBVM ELISA试剂购于上海科华生物制品有限公司；ALT试剂购于北京中生试剂厂.酶标仪为BIOTEK产品EL312E型.全自动生化分析仪为日立7170型.
1.2 方法 sICAM1含量测定采用双抗体夹心ABC-ELISA法，所有样本均在同批检测，严格按说明书操作，依据同批所作标准曲线求出各样品sICAM1含量，比较分析数据.
2 结果
2.1 不同临床类型乙型肝炎患者sICAM1测定结果 见Tab 1.
表1 HBV感染者不同临床类型与正常对照sICAM1含量(略)
Tab 1 Concentrations of sICAM1 compare between control and HBV infecters(略)
由Tab 1可知，急性肝炎组、慢性肝炎组与正常对照组相比sICAM1含量明显升高；无症状携带组与正常对照组无明显差异；自然恢复组与正常对照组无差异.慢性肝炎组中，慢性中度、慢性重度分别于慢性轻度比较，差异均有显著性(P<0.01).
2.2 乙肝病毒感染后病毒复制状态与sICAM1含量的比较 以e抗原作为病毒复制状态的指标.74例HBV感染者中，e抗原阳性者31例，sICAM1含量为(628±239) μg·L-1，e抗原阴性者43例，sICAM1含量为(583±239) μg·L-1，二者相比差异无显著性(P>0.05).
2.3 肝细胞损伤程度与sICAM1含量比较 74例HBV感染者中，ALT大于100者35例，sICAM1含量为(824±390) μg·L-1ALT小于100者39例(包括病毒携带组)sICAM1含量为(386±89) μg·L-1二者差异有显著性(P<0.01)，采用两大样本均数比较的U检验(U＝4.31).
3 讨论
乙肝病毒为嗜肝性DNA病毒，其感染人体后可引起机体对感染的肝细胞的免疫应答，依其应答状态，可引起急性肝炎、慢性肝炎、无症状病毒携带者等不同临床类型，也可发展为肝硬化、肝癌等.早在1990年，Ferrari等报道了急性自限性乙型肝炎，慢性乙型肝炎和无症状表面抗原携带者外周血单个核细胞对乙型肝炎病毒有不同的免疫反应性，他们发现急性乙肝患者Tc应答最强，慢性乙肝和无症状携带者TC细胞对核心抗原几乎没有应答，认为机体对HBcAg 出现强应答，病毒消除快，形成急性自限性乙型肝炎，反之则引起病毒持续性感染.后来，Chisari等[1]采用合成多肽(CD8+T细胞识别位点)也得到类似结果.他们发现CTL作用位点广泛存在于各种病毒抗原的不同区段上，且不同急性乙肝患者因免疫状态各异，CTL作用位点免疫应答强度不同，出现多克隆、多特异性特点.认为机体抗病毒的细胞免疫应答强度可作为急性感染或慢性感染的指标.Volpes的研究发现，乙肝病毒感染时缺乏粘附分子表达的肝细胞不能吸引到白细胞，不能杀伤肝细胞.而且肝细胞粘附分子表达量与白细胞侵润成正比，与临床类型有很好的相关性.由于其研究均采用免疫组化或直接免疫荧光法，肝组织来源不易，给临床应用带来困难.近年来，一些学者采用ELISA法测定患者血清中sICAM1，研究其与肝脏损伤的关系[2-4]，但将其作为判断乙型肝炎临床类型的指标报道甚少.我们测定了74例不同临床类型感染者血清sICAM1水平，结果显示：急性感染及慢性感染者明显高于正常对照组；自然恢复者与正常对照组无明显差异；慢性病毒携带者与正常对照组无明显差异.74例感染者中ALT>100 u·L-1者sICAM1水平显著高于ALT<100 u·L-1者，结果与现有报道一致[5-9]，进一步证实已肝时肝细胞的损伤是由于免疫机制引起的理论.而31例e抗原阳性者， sICAM1含量平均(628±239)μg·L-1，43例e抗原阴性者sICAM1含量平均(583±239) μg·L-1，二者相比,差异无显著性,与He等[2]的结果一致,与LI等[4]的结果相反,有必要扩大样本量以HBV-DNA定量结果继续观察sICAM1含量与病毒复制活跃程度的关系.我们还选择了10例自然恢复者，测定结果sICAM1含量与正常对照组无明显差异.可能的过程为，乙肝病毒进入免疫功能正常的机体的肝细胞，通过一系列的反应，感染的肝细胞将病毒多肽表达于膜表面，同时表达MHCI及ICAM1增加，吸引LFA1阳性淋巴细胞，通过抗原受体及分子配体间的结合，使大量的淋巴细胞活化，攻击靶细胞，清除病毒.随着病毒的清除，ICAM1表达减少或消失，病变恢复[10-13].综上所述，血清中sICAM-1测定可作为判断乙型肝炎临床类型的指标；感染者血清中sICAM-1含量明显升高，提示急性乙型肝炎或慢性肝炎；如果血清中sICAM1含量不高,提示病毒感染可能趋于恢复或提示可能为免疫耐受模型,为无症状病毒携带者.sICAM1含量与病毒复制活跃程度无关，与肝脏损伤程度成正比.
【参考文献】
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[8] Zhang XQ, Zhang R, Gu ZH, Wang YM. Study on the expression of ICAM1and HLAA, B, C in the liver of viral hepatitis B [J]. Disan Junyi Daxue Xuebao (J Fourth Mil Med Univ), 2001 ; 23(7): 841-843.
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[10] Zhang XQ, Gu ZH. Expression of ICAM1 in the liver of patients with hepatitis B[J]. Zhonghua Ganzhangbing Zazhi (Chin J Hepatol), 1998;6:206-207.
[11] Stancek D, Fuchsberger N, Oltman M, Schmeisser H, Kontsek P, Jahnova E, Hajnicka V. Significance of antiinterferonalpha2 and sICAM1 activities in the sera of viral hepatitis B and C patients treated with human recombinant interferonalpha2 [J]. Acta Virol, 2001;45(5-6):287-292.
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[13] Spahr Laurent, Rubbia Brandt, Laura, Pugin Jerome, Giostra Emile, Frossard Jean Louis. Rapid changes in alcoholic hepatitis histology under steroids: Correlation with soluble intercellular adhesion molecule1 in hepatic venous blood [J]. J Hepatol,2001; 35(3): 582-589.
编辑 袁天峰
作者简介：王香玲(1957)，女(汉族)，陕西省高陵县人，副主任检验师. Tel.(029)7276936 Email.wenning@ahoo.com.cn., http://www.100md.com(王香玲，王小利，赵丽华，李妙羡，张西英 谢明，纪玉)
Significance of serum sICAM1 in the clinical types of hepatitis B
WANG XiangLing, WANG XiaoLi, ZHAO LiHua, LI MiaoXian, ZHANG XiYing， XIE Ming, JI YuQiang
Department of Clinical Laboratory, Second Hospital, Xian Jiaotong University，Xian 710004，China，Department of Immunology，Medical College, Xian Jiaotong University，Xian 710061, China
【Abstract】 AIM: To observe the changes and significance of serum sICAM1 in the assessing clinical types of Hepatitis B. METHODS: Double antibody sandwich ABCELISA was employed for the quantitative measurement of serum sICAM1 of 74 cases of different clinical types and 10 cases of normal control . RESULTS: Both the sICAM1 level of 13 cases of acute hepatitis B (680±248) μg·L-1 and that of 46 cases of chronic hepatitis B (675±311) μg·L-1 were significantly higher than that of normal control (204±45) μg·L-1. The sICAM1 level of 18 cases of moderate chronic hepatitis B (634±182) μg·L-1 was significantly higher than the 19 cases of mild chronic hepatitis B (445±120) μg·L-1. The sICAM1 level of 9 cases of serious chronic hepatitis B (931±280) μg·L-1 was significantly higher than that of the group of mild chronic hepatitis B. The sICAM1 level of 10 cases of completely recovered cases (243±86) μg·L-1 and that of 5 cases of asymptomtic carriers (212±72) μg·L-1 were not significantly different from that of the normal control. Serum concentrations of sICAM1 were significantly positively correlated with the concentrations of ALT. There was no significant difference between the HBeAg positive group and HBeAg negative group. CONCLUSION: Detection of sICAM1 can be used as a indicator in assessing clinical types. The concentration of sICAM1 is positivley correlated with the level of liver damage but is not related to the virus duplication.
【Keywords】 sICAM1; Hepatitis B; clinical types
【摘 要】目的: 观察血清sICAM-1水平在HBV感染者不同临床类型中的变化及其意义.方法 ： 采用双抗体夹心ABC-ELISA法，对74例不同临床类型HBV感染者及10例正常对照者血清中可溶性细胞间粘附分子-1(sICAM1)进行了定量测定. 结果 ： 急性肝炎13例血清中sICAM-1含量(680±248) μg·L-1明显高于正常对照组(204±46) μg·L-1 (P<001); 46例慢性肝炎患者血清中sICAM-1含量(675±311) μg·L-1明显高于正常对照组(P<001);其中慢性肝炎轻度19例，sICAM-1含量(445±120) μg·L-1；慢性肝炎中度18例，sICAM-1含量为(634±182)μg·L-1，明显高于慢性轻度组(P<001)；慢性肝炎重度9例，sICAM1含量(931±280)μg·L-1，明显高于慢性轻度组(P<001).5例无症状携带者sICAM1含量(212±72) μg·L-1，与正常对照组差别无显著性(P>005); 10例自然恢复组含量为(243±86) μg·L-1，与正常对照组差别无显著性.血清中sICAM1含量与ALT含量呈明显正相关;而E抗原阳性组与E抗原阴性组差异无显著性.结论： 血清中sICAM-1水平测定可作为判别乙型肝炎临床类型的指标；sICAM1含量与肝脏损伤程度成正比，sICAM1含量与病毒复制状态无关.
【关键词】 sICAM1；肝炎乙型，临床类型
0 引言
乙型肝炎为全球性传染病,根据病程可分为急性乙型肝炎, 慢性乙型肝炎.根据临床表现可分为轻型、中型、重型乙型肝炎和无症状乙肝病毒携带者.可发展为肝硬化、肝癌.目前尚无确切可靠的特异治疗方法,也无提示其预后的明确指标.近年来大量研究认为, 乙型肝炎不同的临床类型与机体抗病毒免疫应答密切相关.ICAM1能介导免疫反应中各类细胞间的粘附，目前研究认为，ICAM1/LFA1介导的细胞粘附在炎症性肝病时T细胞与肝细胞相互作用中起着关键性作用.为观察乙型肝炎不同临床表现者细胞间黏附分子的变化,我们测定了74例不同临床类型乙肝感染者血清中sICAM1水平, 报告如下.
1 对象和方法
1.1 对象 乙肝患者59(男45，女14)例，年龄7~51岁，其中急性肝炎13例，慢性肝炎轻度19例，中度18例，重度9例，均为我院传染科住院或门诊患者，用免疫制剂治疗前采集标本；无症状携带者5(男4，女1)例，年龄16~46岁，为门诊复检者，检出HBsAg阳性超过1a；自然恢复者10(男6，女4)例，年龄15~52岁，为门诊复检者，HBVM检测为抗HBc和抗HBs阳性或抗HBe，抗HBc，抗HBs阳性或抗HBs阳性，未注射乙肝疫苗；正常对照10(男7，女3)例，年龄15~55岁，均为门诊健康体检者，未注射乙肝疫苗.常规静脉采血，4 h内分离血清，检测乙肝病毒标志物及ALT，同时分出血清于-30℃保存.sICAM1试剂购于上海森雄公司；HBVM ELISA试剂购于上海科华生物制品有限公司；ALT试剂购于北京中生试剂厂.酶标仪为BIOTEK产品EL312E型.全自动生化分析仪为日立7170型.
1.2 方法 sICAM1含量测定采用双抗体夹心ABC-ELISA法，所有样本均在同批检测，严格按说明书操作，依据同批所作标准曲线求出各样品sICAM1含量，比较分析数据.
2 结果
2.1 不同临床类型乙型肝炎患者sICAM1测定结果 见Tab 1.
表1 HBV感染者不同临床类型与正常对照sICAM1含量(略)
Tab 1 Concentrations of sICAM1 compare between control and HBV infecters(略)
由Tab 1可知，急性肝炎组、慢性肝炎组与正常对照组相比sICAM1含量明显升高；无症状携带组与正常对照组无明显差异；自然恢复组与正常对照组无差异.慢性肝炎组中，慢性中度、慢性重度分别于慢性轻度比较，差异均有显著性(P<0.01).
2.2 乙肝病毒感染后病毒复制状态与sICAM1含量的比较 以e抗原作为病毒复制状态的指标.74例HBV感染者中，e抗原阳性者31例，sICAM1含量为(628±239) μg·L-1，e抗原阴性者43例，sICAM1含量为(583±239) μg·L-1，二者相比差异无显著性(P>0.05).
2.3 肝细胞损伤程度与sICAM1含量比较 74例HBV感染者中，ALT大于100者35例，sICAM1含量为(824±390) μg·L-1ALT小于100者39例(包括病毒携带组)sICAM1含量为(386±89) μg·L-1二者差异有显著性(P<0.01)，采用两大样本均数比较的U检验(U＝4.31).
3 讨论
乙肝病毒为嗜肝性DNA病毒，其感染人体后可引起机体对感染的肝细胞的免疫应答，依其应答状态，可引起急性肝炎、慢性肝炎、无症状病毒携带者等不同临床类型，也可发展为肝硬化、肝癌等.早在1990年，Ferrari等报道了急性自限性乙型肝炎，慢性乙型肝炎和无症状表面抗原携带者外周血单个核细胞对乙型肝炎病毒有不同的免疫反应性，他们发现急性乙肝患者Tc应答最强，慢性乙肝和无症状携带者TC细胞对核心抗原几乎没有应答，认为机体对HBcAg 出现强应答，病毒消除快，形成急性自限性乙型肝炎，反之则引起病毒持续性感染.后来，Chisari等[1]采用合成多肽(CD8+T细胞识别位点)也得到类似结果.他们发现CTL作用位点广泛存在于各种病毒抗原的不同区段上，且不同急性乙肝患者因免疫状态各异，CTL作用位点免疫应答强度不同，出现多克隆、多特异性特点.认为机体抗病毒的细胞免疫应答强度可作为急性感染或慢性感染的指标.Volpes的研究发现，乙肝病毒感染时缺乏粘附分子表达的肝细胞不能吸引到白细胞，不能杀伤肝细胞.而且肝细胞粘附分子表达量与白细胞侵润成正比，与临床类型有很好的相关性.由于其研究均采用免疫组化或直接免疫荧光法，肝组织来源不易，给临床应用带来困难.近年来，一些学者采用ELISA法测定患者血清中sICAM1，研究其与肝脏损伤的关系[2-4]，但将其作为判断乙型肝炎临床类型的指标报道甚少.我们测定了74例不同临床类型感染者血清sICAM1水平，结果显示：急性感染及慢性感染者明显高于正常对照组；自然恢复者与正常对照组无明显差异；慢性病毒携带者与正常对照组无明显差异.74例感染者中ALT>100 u·L-1者sICAM1水平显著高于ALT<100 u·L-1者，结果与现有报道一致[5-9]，进一步证实已肝时肝细胞的损伤是由于免疫机制引起的理论.而31例e抗原阳性者， sICAM1含量平均(628±239)μg·L-1，43例e抗原阴性者sICAM1含量平均(583±239) μg·L-1，二者相比,差异无显著性,与He等[2]的结果一致,与LI等[4]的结果相反,有必要扩大样本量以HBV-DNA定量结果继续观察sICAM1含量与病毒复制活跃程度的关系.我们还选择了10例自然恢复者，测定结果sICAM1含量与正常对照组无明显差异.可能的过程为，乙肝病毒进入免疫功能正常的机体的肝细胞，通过一系列的反应，感染的肝细胞将病毒多肽表达于膜表面，同时表达MHCI及ICAM1增加，吸引LFA1阳性淋巴细胞，通过抗原受体及分子配体间的结合，使大量的淋巴细胞活化，攻击靶细胞，清除病毒.随着病毒的清除，ICAM1表达减少或消失，病变恢复[10-13].综上所述，血清中sICAM-1测定可作为判断乙型肝炎临床类型的指标；感染者血清中sICAM-1含量明显升高，提示急性乙型肝炎或慢性肝炎；如果血清中sICAM1含量不高,提示病毒感染可能趋于恢复或提示可能为免疫耐受模型,为无症状病毒携带者.sICAM1含量与病毒复制活跃程度无关，与肝脏损伤程度成正比.
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编辑 袁天峰
作者简介：王香玲(1957)，女(汉族)，陕西省高陵县人，副主任检验师. Tel.(029)7276936 Email.wenning@ahoo.com.cn., http://www.100md.com(王香玲，王小利，赵丽华，李妙羡，张西英 谢明，纪玉)