慢性乙型肝炎肝内浸润淋巴细胞功能相关抗原-1表达在肝细胞损伤及病毒清除中的意义

探讨淋巴细胞功能相关抗原—1(LFA-1）在慢性乙肝组织中浸润淋巴细胞的表达及意义。利用免疫组化技术，对51例不同类型的慢性HBV感染者肝组织内浸润淋巴细胞LFA-1的表达状况进行观察，并同时检测肝内HBcAg表达状况。慢性HBV感染者肝内浸润淋巴细胞存在不同程度的LFA-1表达，9例HBV感染的无症状携带者，肝内散在的淋巴细胞未见LFA-1表达，在42例慢性乙性肝炎中，其中38例肝组织浸润淋巴细胞LFA-1表达，阳性率为90．4％。阳性表达的淋巴细胞多位于肝组织病变明显区域及其周围肝组织，LFA-1表达强度与肝炎的程度有关，其中慢性重度乙型肝炎浸润淋巴细胞LFA—1表达较慢性中度乙型肝炎显著增强，慢性中度乙型肝炎LFA-1表达较慢性轻度乙型肝炎显著增强。肝内浸润淋巴细胞LFA-1表达阳性组、强阳性组较弱阳性及阴性组相比，肝内HBcAg阳性表达减少。慢性乙肝组织中浸润淋巴细胞LFA-1的表达参与了乙型肝炎的发病过程，不仅参与肝细胞的损害过程，同时亦有助于对肝细胞内HBV的清除。     

用单克隆抗HBcAg抗体对慢性活动型肝炎时HBcAg的形态学观察—并与多克隆抗体比较

用单克隆抗体研究乙型肝炎时HBV的几种抗原在肝组织内的定位,形态学及其与肝组织病变和临床表现的相互关系,国内正在开展中。本文对乙型肝炎慢性活动型46例肝组织用单克隆抗HBcAg抗体,ABC法作组织化学检测,并与多克隆人抗HBcAg抗体检测相比较。单克隆抗体染色背景清晰,非特异性染色低,提高了染色质量。本株单克隆抗体检测之阳性率与多克隆抗体基本一致。HBcAg阳性细胞可表现为胞核型,胞     

乙型肝炎患者肝组织中Fas及Fas配体的表达

为探讨乙型肝炎肝组织中Ｆａｓ及Ｆａｓ配体（ＦａｓＬ）表达和分布的相互关系，采用免疫组织化学技术，以兔抗－Ｆａｓ及兔抗－ＦａｓＬ多克隆抗体，对６０例急性轻型肝炎、慢性活动性肝炎及活动性肝硬化患者石蜡包埋肝组织中的Ｆａｓ及ＦａｓＬ进行了检测。Ｆａｓ及ＦａｓＬ的检出率分别为７６．７％（４６／６０）及７０．０％（４２／６０），Ｆａｓ于肝细胞胞浆内表达，ＦａｓＬ多在肝组织中浸润的淋巴细胞胞浆内表达（３４／４２，８０．９％），也见于肝细胞胞浆中（２５／４２，５９．５％）。ＦａｓＬ阳性淋巴细胞主要分布于汇管区，肝小叶内很少见，ＦａｓＬ阳性肝细胞的分布类同Ｆａｓ阳性肝细胞；在急性轻型肝炎，阳性细胞多在小叶内弥散分布，在慢性活动性肝炎和活动性肝硬化则更多集聚于碎屑样坏死灶和假小叶的周边。免疫组化双标记染色显示，Ｆａｓ及ＦａｓＬ可在同一或不同肝细胞胞浆内表达，但多分布于同一区域。本研究在正常肝组织中未查见ＦａｓＬ的表达，提示在病理状态下，肝细胞才出现ＦａｓＬ的表达。Ｆａｓ及ＦａｓＬ在肝组织中的分布表明，Ｆａｓ－ＦａｓＬ系统在乙型病毒性肝炎肝细胞损伤中起着重要的作用。     

慢性乙肝患者肝脏炎症活动与肝组织HBcAg表达的相关性分析

探讨慢性乙型肝炎患者肝脏炎症活动与肝组织HBcAg表达的相关性。随机选择102例住院慢性乙型肝炎患者,肝穿活检,行HE染色及免疫组化染色,光镜观察其肝组织炎症活动与免疫组化HBcAg染色阳性率的关系。随慢性乙肝患者肝组织炎症活动度的增加,各亚型HBcAg阳性率有明显差异,P〈0．05。慢性乙肝患者肝组织炎症活动度与HBcAg阳性亚型有关。     

珠子肝泰治疗慢性乙型肝炎的免疫组织化学研究

目的：观察珠子肝泰对慢性乙型肝炎患者肝组织内HBsAg和HBcAg的影响。方法：对21例慢性乙型肝炎应用珠子肝泰进行治疗，3次／d，每次4粒（0．8g)，疗程6个月。 在治疗开始前和疗程结束后采集血清和肝细胞标本，进行血清学、病理学和免疫组织化学检查。结果：治疗后、血清内HBeAg和HBV DNA的转阴率分别为45．0％和47．4％，81％的患者肝组织损伤改善或修复，45．4％的患者肝纤维化程度减轻，肝内HBsAg和HBcAg的转阴率分别为19．0％和23．8％。结论：珠子肝泰在抑制慢性乙型肝炎患者血清HBV复制标志和改善肝组织病理损伤程度的同时，对肝细胞内乙肝病毒感染因子也有一定抑制作用。     

丁型肝炎患者肝组织HDAg和HBsAg/HBcAg的免疫组化检测

目的 探讨HDAg和HBsAg/HBcAg在丁型肝炎患者肝组织中表达及关系。方法 应用免疫组化双重染色,检测79例丁型肝炎患者肝组织HDAg、HBsAg和HBcAg表达,以52例乙型肝炎作对照。结果 丁型肝炎HBsAg、HBcAg检出率(81％、71％)较乙型肝炎(94％、92％)低(P<0.05或0.01)。HDAg以肝细胞核表达为主,HBsAg以肝细胞浆表达为主,HDAg和HBsAg表达强度及阳性细胞分布呈一致性,且均与肝组织的炎症活动和病理损害程度相关(P<0.01)。HBcAg以肝细胞核表达为主,阳性细胞主要呈单个细胞或点状分布,且HBcAg阳性细胞明显少于HDAg阳性细胞。结论 HDV感染会抑制HBV病毒抗原(HBcAg)表达;HDV致病机制中既有HDV的直接细胞毒性作用,也有HBV和HDV的协同作用。     

慢性肝病患者肝组织中庚型肝炎病毒抗原的定位研究

应用抗HGV NS5单克隆抗体(McAb),以辣根过氧化物酶与抗辣根过氧化物酶法(PAP)检测了166例慢性肝病患者(CLD)肝组织中庚型肝炎病毒抗原(HGV-Ag)。结果显示,慢性非甲～戊型肝炎、慢性丙型肝炎、慢性乙型肝炎、肝硬变和肝细胞癌患者阳性检出率分别为28.57％、22.45％、14.28％、18.52％和15.38％。HGV-Ag阳性肝细胞多呈散在、局灶和弥漫性三种形式分布,HGV-Ag染色阳性颗粒可见于细胞浆中,HGV-Ag NS5的检出率与HBV、HCV感染有关。提示HGV NS5抗原在慢性肝病患者肝细胞中表达,且HGV NS5的表达与血清HGV RNA水平关系较为密切,HGV感染可能在慢性肝病发生中起一定病原学作用。     

重型乙型肝炎肝内细胞间粘附分子—1 mRNA的表达

目的 探讨细胞间粘附分子－１（ＩＣＡＭ－１）ｍＲＮＡ在重型乙型肝炎肝内的表达水平及其意义。方法 用原位杂交技术检测１１例非肝病对照者,５例慢性无症状ＨＢｓＡｇ携带者（ＡｓＣ）,１５例慢性乙型肝炎和３０例重型乙型肝炎患者肝内ＩＣＡＭ－１ ｍＲＮＡ原位表达。结果 非肝病者和ＡｓＣ肝细胞无ＩＣＡＭ－１ ｍＲＮＡ;重型乙型肝炎患者肝细胞ＩＣＡＭ－１ ｍＲＮＡ表达明显增强,其表达水平显著高于慢性乙型肝炎。结     

细胞间粘附分子－1在乙型肝炎中的表达及意义

为观察细胞间粘附分子－１（ＩＣＡＭ－１）在乙型肝炎肝组织内的表达状况，探讨ＩＣＡＭ－１在乙型肝炎肝细胞免疫损伤中的作用。采用链菌素亲生物蛋白法（ＬＳＡＢ），以ＩＣＡＭ－１的单克隆抗体，检测１０４例急、慢性乙型肝炎，９例无症状携带者，１０例正常肝组织标本内ＩＣＡＭ－１抗原表达情况。结果发现：ＩＣＡＭ－１在正常肝细胞无表达，肝窦内皮细胞弱表达；ＨＢＶ感染后肝细胞呈不同程度的表达，肝窦内皮表达增强；中、重度慢性肝炎，活动性肝硬化肝细胞ＩＣＡＭ－１表达较急性肝炎，轻度慢性肝炎显著增强（Ｐ＜０．０１）；急性肝炎较轻度慢性肝炎显著增强（Ｐ＜０．０１）；轻度慢性肝炎较无症状携带者，正常肝组织显著增强（Ｐ＜０．０１）。表明ＩＣＡＭ－１在乙型肝炎肝细胞内表达与肝细胞损伤有关，对ＨＢＶ的清除可能起着重要作用，ＩＣＡＭ－１在肝窦内皮的表达有助于淋巴细胞向肝组织内浸润。     

慢性乙型肝炎患者肝脏炎症活动度与肝组织HBcAg免疫组化的关系初探

目的：探讨慢性乙型肝炎患者肝脏炎症活动度与肝组织HBcAg表达的相关性。方法：随机选择102例慢性乙型肝炎患者，肝穿刺活检，行HE染色及免疫组化染色，光镜观察肝脏炎症活动度与肝组织HBcAg各亚型表达的关系。结果：随慢性乙型肝炎患者肝脏炎症活动度的增加，各亚型HBcAg阳性率差异有显著性意义（P〈0．05）。结论：慢性乙型肝炎患者肝脏炎症活动度与肝组织HBcAg免疫组化阳性亚型有关。     

乙型肝炎病毒核心抗原在慢性乙型肝炎病毒感染者肝细胞内的分布及临床意义

目的 探讨HBcAg在慢性HBV感染者肝细胞内的分布情况及其临床意义.方法 对41例慢性HBV感染者进行肝组织穿刺,用免疫荧光组织化学技术在激光共聚焦显微镜下观察肝细胞内HBcAg的分布情况.计量资料采用Kruskal Wallis检验,计数资料采用卡方检验.结果 41例慢性HBV感染者中,36例肝细胞内HBcAg阳性表达,阳性率为87.8%,其中23例肝功能中度异常,10例肝功能轻度异常,3例肝功能正常.HBcAg有膜、胞质与核三种形式表达.在23例肝功能中度异常者中,6例呈明显膜型表达,无明显胞质型及核型,17例以胞质型与膜型混合表达,未发现核型表达.在10例肝功能轻度异常者中,以单纯胞质型为主,未见膜型与核型表达.在3例肝功能正常者中,以胞质型表达与少许核型表达为主,未见膜型表达.HBcAg表达类型与肝功能之间差异有统计学意义(χ2=10.60,P＜0.01).结论 慢性HBV感染者肝组织损伤与HBcAg的表达有直接联系,提示膜型表达的HBcAg是肝脏免疫损伤过程中的靶抗原.

Abstract：

Objective To explore the distribution and clinical significance of hepatitis B core antigen( HBcAg) in the hepatocytes of chronic hepatitis B virus infected patients. Methods Paraffin sections were made from 41 liver biopsy samples obtained from chronic hepatitis B virus infected patients. The immuno-fluorescence confocal technique was utilized to analyze the expression level and localization of HBcAg in hepatocytes. The data were analyzed by using Kruskal Wallis test and chisquare test. Results HBcAg expression were detected in 36 (87. 8%) patients, among whom 23 cases had moderate abnormal liver function, 10 with mild abnormal liver function and 3 with normal liver function. Among the cases with moderate abnormal liver function, 6 cases showed the simple membrane-type HBcAg expression, 17 cases showed mixed cytosolic-type and membrane-type HBcAg expression without the nuclear-type expression. Twelve cases with mild abnormal liver function only showed simple cytosolic-type HBcAg expression without membrane-type or nuclear-type expression. In the three patients with normal liver function, HBcAg was expressed in cytoplasm and nuclear but not on membrane. The correlation between HBcAg expression pattern and liver function was statistically significant (χ2 =10. 60, P＜0.01). Conclusion HBcAg expression is directly correlated with liver injury in chronic hepatitis B virus infected patients, which indicates that membrane expressed HBcAg is the target antigen during the immuno-attack of liver.     

Liver disease activity and hepatitis B virus replication in chronic delta antigen-positive hepatitis B virus carriers

Delta antigen is currently thought to reflect superinfection of the liver with a defective RNA virus (delta agent), requiring helper function from hepatitis B virus for its replication. To assess the influence of delta agent on hepatitis B virus replication in patients persistently infected with both viruses and showing chronic liver disease, we measured serum and liver hepatitis B virus DNA in HBsAg-positive chronic liver disease patients who were either positive or negative for delta antigen in the liver. Hepatitis B virus DNA was assayed in the serum of 21 patients with delta antigen-positive/HBsAg-positive chronic liver disease and in 21 patients with delta antigen-negative/HBsAg-positive chronic liver disease matched for HBeAg/anti-HBe status and underlying liver histology. HBcAg and delta antigen in liver was determined by immunofluorescence or immunoperoxidase staining. In delta antigen-positive/HBsAg-positive chronic liver disease, serum hepatitis B virus DNA was detected transiently in 4 of 21 cases (19%) and was present in these patients at low levels (trace to 2+). In contrast, 9 of 21 (43%) delta antigen-negative/HBsAg-positive chronic liver disease patients were serum hepatitis B virus DNA positive, and five of these had high serum hepatitis B virus DNA levels (3+ to 4+). Serum HBsAg and anti-HBc titers were significantly lower in delta antigen-positive cases and correlated with reduced amount of HBcAg in the liver.(ABSTRACT TRUNCATED AT 250 WORDS)     

Significance of hepatitis B core antigen in the liver in patients with chronic hepatitis B and its relation to hepatitis B virus DNA

Liver biopsies from 52 patients with chronic hepatitis B were investigated for the presence and distribution of HBcAg and the results were compared with the status of hepatitis B virus deoxyribonucleic acid (HBV-DNA). The patients consisted of 37 men and 15 women, aged 16-55 years (mean = 34 years). Serum alanine aminotransferase (ALT) was elevated in 50 patients (range: 18-969 U/L; mean = 290 U/L). Serological testing showed HBsAg in all, HBeAg in 45 (87%), and HBV-DNA in 28 (54%). Liver biopsies demonstrated HBcAg in 35 (67%) patients. HBcAg was not only present in 31 of 45 (69%) patients who were seropositive for HBeAg, but also in four of seven (57%) with antibody to HBeAg (anti-HBe). In 28 of 35 (80%) patients with HBcAg in the liver, serum HBV-DNA was detected. However, no serum HBV-DNA was detected in 17 patients who had no detectable HBcAg in the liver. The distribution of HBcAg in the liver was rather cytoplasmic and nuclear than nuclear alone. Among 33 patients with cytoplasmic HBcAg in the liver, 15 (45%) had an evidence of acute exacerbation of hepatitis with marked ALT elevation (range: 168-894 U/L; mean = 385 U/L) and nine patients showed severe chronic active hepatitis and confluent necrosis, histologically. These results indicate that the presence of HBcAg in the liver correlates with the amount of circulating hepatitis B virus as quantified by serum level of HBV-DNA. The predominant cytoplasmic HBcAg in the liver may suggest the possibility of multiple episodes of acute exacerbation and more severe ongoing hepatitis during the clinical course.     

Cryptogenic chronic liver disease and serum or liver hepatitis B virus markers. Their possible correlations and etiologic significance

In an attempt to evaluate a possible correlation between cryptogenic chronic liver disease and a present or past hepatitis B virus (HBV) infection, we studied 17 patients with hepatitis B surface antigen (HBsAg)-negative, nonalcoholic chronic liver disease; 9 of them were positive for serum HBsAg detected by a solid-phase enzyme immunoassay with monoclonal antibody (M-EIA) and 8 were negative for the same marker. Liver hepatitis B core antigen (HBcAg), studied by an indirect immunofluorescence technique, was present in 55.5% of the patients positive for serum HBsAg by M-EIA. In the same group of patients, liver HBV-DNA was found in 66.6% of the patients. On the other hand, only 1 patient without serum positivity for HBsAg by M-EIA was positive for liver HBcAg and HBV-DNA. None of our patients showed serum positivity for HBV-DNA sequences. We conclude that HBV infection may be a possible cause of cryptogenic chronic liver disease; this HBV-related, HBsAg-negative chronic liver disease seems to have no viral replication or undetectable levels of HBV-DNA in serum. HBsAg, detected by a monoclonal assay, seems to be a suitable marker to identify this subgroup of patients with HBsAg-negative chronic liver disease.     

单项抗-HBs阳性肝炎的临床与病理分析

目的 分析单项抗-HBs阳性肝炎的临床与病理特征,提高对单项抗-HBs阳性肝炎的认识。方法 对72例单项抗-HBs阳性肝炎患者进行肝炎病毒血清学检测、肝功能检查、血清HBV DNA检测及肝穿刺活组织病理和免疫组化检查。结果 72例患者除去4例脂肪肝和3例Dubin-Johnson综合征外,另65例中89.2％为慢性肝炎(58/65),其中慢性肝炎轻度占慢性肝炎79.3％(46/58)。与病理诊断比较临床诊断的正确率为61.1％,在慢性肝炎轻度则较高为71.7％。临床表现和肝功能变化与肝组织炎症活动度密切相关。3.2％的患者示肝内HBsAg或/和HBcAg阳性,而血清HBV DNA检测7.9％阳性。结论 单项抗-HBs阳性肝炎多为慢性肝炎,临床表现轻,炎症活动度和纤维化程度较轻,但体内仍可有HBV复制,仍可发展为肝硬化。肝穿刺病理及免疫组化检测有助于明确诊断。     

隐匿性乙型肝炎病毒感染者临床特点和肝组织病理学检查

目的了解血清肝炎病毒标志物阴性、肝功能反复异常患者中HBV隐匿性感染的比例及其临床和病理学特点。方法对27例血清肝炎病毒标志物阴性、肝功能反复异常患者采用免疫组化法检测肝组织HBsAg、HBcAg和HCVAg，并进行常规的病理学检查。结果肝组织HBsAg和（或）HBcAg阳性9例（33．3％）；HBsAg和（或）HBcAg及HCVAg阳性10例（37．0％）；全阴性8例（29．6％）。在HBV隐匿性感染的19例患者中，慢性肝炎8例，肝硬化11例。结论HBV和HCV感染为血清肝炎病毒标志物阴性患者肝功能反复异常的主要原因之一，尤其是HBV感染。这种HBV隐匿性感染与慢性肝炎、肝硬化的发生关系密切，应引起重视。     

Differential distribution of hepatitis B surface antigen and hepatitis B core antigen in the liver of hepatitis B patients.

One hundred liver biopsies from 100 patients with clinical presumptive diagnosis of hepatitis were examined by immunofluorescence for the presence of hepatitis B surface antigen (HBSAg) and hepatitis B core antigen (HBcAg). Of the 60 HBsAg-positive livers, 51 were diagnosed as chronic hepatitis on histological grounds, 6 as acute hepatitis, and 3 as "near-normal liver." From the 60 tissue-positive cases, 3 subjects were HBsAg seronegative. HBcAg was detected in 44 livers, all of which also had HBcAg in the localized in the cytoplasm and the membranes of the hepatocytes, and HBcAg in the nuclei and in 4 cases also in the cytoplasm. Predominant HBsAg expression in the cytoplasm was observed in near-normal liver, chronic persistent hepatitis, and cirrhosis with little activity. This correlated with the amount of ground glass hepatocytes in the biopsies. HBcAg and membrane-localized HBsAg were minimal in those conditions. HBcAg was most prevalent in patients with chronic aggressive hepatitis and active cirrhosis treated with immunosuppressive drugs, whereas the amounts of HBsAg and HBcAg in nontreated patients of those two groups and in acute hepatitis with signs of transition to chronicity were almost equal. HBsAg expression in liver cell membranes was most prominent in active forms of chronic hepatitis (chronic aggressive hepatitis and in active cirrhosis) and in acute hepatitis with signs of transition to chronicity. This observation correlated in the presence of HBcAg in the biopsies of those patients. In acute hepatitis both HBsAg and HBcAg were detected rarely and no membrane expression of HBsAg was observed. The over-all results show a significant relationship between the different degrees of accumulation of HBsAg and HBcAg in the liver and the various histological types of hepatitis and further suggest an interplay of both hepatitis B virus and host immune response in the development and pathogenesis of hepatitis B.     

Biologic and prognostic significance of hepatocyte hepatitis B core antigen expressions in the natural course of chronic hepatitis B virus infection

To elucidate the biologic significance of hepatocyte hepatitis B core antigen (HBcAg) expression and its relation to the natural course of hepatitis B virus (HBV) infection, the patterns of HBcAg were correlated with HBV virus replication state and the disease activity in 598 needle liver biopsies performed on 569 hepatitis B surface antigen (HBsAg) carriers aged 1-81 years. A good correlation of liver HBcAg with serum HBeAg and HBV DNA status was demonstrated. HBcAg was present in the hepatocyte nuclei (nHBcAg) or cytoplasm (cHBcAg), or in both (mixed). Pure nHBcAg was seen mainly in children and young adults; 86% of the patients had non-aggressive disease, but rare cases of chronic active hepatitis (CAH) and HBeAg seroconversion were observed. In contrast, cHBcAg was predominantly associated with CAH (52%) and accompanied by a significantly higher HBeAg seroconversion rate (27%). The HBeAg-negative group, particularly the liver HBcAg-negative subgroup, had a lower frequency of CAH, but an increased incidence of non-aggressive disease as well as cirrhosis and/or hepatocellular carcinoma, indicating that HBeAg seroconversion to anti-HBe does not necessarily mean a favorable prognosis. The results suggest that expression of HBcAg correlates with the liver pathology and the three phases of chronic HBV infection: (1) the early immune tolerance phase is characterized by nHBcAg, mild disease and low HBeAg seroconversion rate; (2) the virus replication/elimination phase by cHBcAg or negative HBcAg, frequent CAH, and high HBeAg seroconversion rate; and (3) the inactive virus replication phase by negative HBcAg and a bipolar disease spectrum.     

Hepatitis D virus (delta agent) superinfection in an endemic area of hepatitis B infection: immunopathologic and serologic findings

In 86 Chinese patients with histologically proven hepatitis B surface antigen (HBsAg) positive chronic hepatitis and serum alanine aminotransferase levels exceeding 200 U/l, antibody to hepatitis D antigen (HDAg) was detected more frequently in sera from hepatitis B e antigen (HBeAg) negative patients (11/35, 31.4%) than in HBeAg positive (4/51, 7.8%) patients (p less than 0.02). 10 liver biopsy specimens (76.9%) from 13 chronic hepatitis B patients with superimposed hepatitis D virus (HDV) infection, showed positive staining for HDAg in their hepatocytes. Neither HBsAg nor hepatitis B core antigen (HBcAg) was found in the liver in 12/13 patients with superimposed HDV infection. However, in liver biopsy specimens from 42 patients without HDV superinfection, HBsAg was stained positively in 41 patients (97.6%), and HBcAg in 24 patients (47.1%). Using dot blot hybridization technique, serum hepatitis B virus (HBV) DNA was detected in 62.1% (41/66) of patients without HDV superinfection, while it was detected only in 10.0% (1/10) of patients who had HDV superinfection. It is concluded that HDV superinfection plays a significant role in Taiwan in HBeAg negative chronic hepatitis B patients with clinical "exacerbation". The data show clear evidence of HDV interfering with the replication of HBV.