Michalak T I, Lin B, Churchill N D, Dzwonkowski P, Desousa J R
Liver Research Laboratory, Division of Basic Medical Sciences, Faculty of Medicine, Memorial University of Newfoundland, St. John's, Canada.
Lab Invest. 1990 Jun;62(6):680-9.
Hepatocyte plasma membranes purified from five woodchucks with distinct serologic and histologic patterns of experimentally induced acute woodchuck hepatitis virus (WHV) infection were studied to determine the virus antigens expression and anti-viral specificity of the bound immunoglobulins. WHV core, e, and surface antigens (WHcAg, WHeAg, and WHsAg, respectively) were analyzed with the use of immunoblotting technique both in the native form of these membranes and in the membranes treated with high molar urea or a nonionic detergent. The eluted material was tested either for the presence of WHV antigens or reactivity of the antibodies directed to the virus antigens. The data revealed that acute WHV infection is accompanied by hepatocyte plasma membrane expression of all three viral antigens tested. In all cases, native membranes displayed both WHeAg and WHsAg, whereas WHcAg presence was detected in hepatocyte plasma membranes after their disruption with urea or a detergent. The data indicated that a part or, in some instances, even the whole detectable WHcAg specificity can be incorporated into plasma membrane structure in such a way that it is not accessible for recognition by the specific antibodies (anti-WHc), suggesting at least a partial functional disability of this antigen as a target for immunologic reactions in in vivo conditions. In contrast, WHeAg specificity was detectable in all native membrane preparations studied and its expression was not evidently influenced by the employed treatments, whereas that of WHsAg tended to decline. Further, anti-WHc reactivity was identified in all membrane eluates tested, but antibodies to WHeAg (anti-WHe) were exclusively found in the material eluted from membranes originating from woodchucks with borderline histologic activity of acute hepatitis, which cleared away e antigen from the serum shortly before liver perfusion. Antibodies to WHsAg (anti-WHs) did not show up in the eluates. The present findings demonstrated that WHeAg specificity is not only exposed on the surface of infected hepatocytes, but is also relatively more easily accessible for serologic recognition than that of WHcAg in acute WHV infection. The above observation suggests that e antigen can serve as a potential plasma membrane target for hepatocytolytic attack in addition to that of WHsAg or WHcAg. Moreover, the results of this study demonstrated an apparent relationship between low histologic activity of liver inflammation, e antigen clearance from the circulation, and detectability of hepatocyte plasma membrane-bound anti-e antibodies in acute hepadna viral hepatitis.
从五只经实验诱导感染土拨鼠肝炎病毒（WHV）且具有不同血清学和组织学模式的土拨鼠中纯化肝细胞质膜，以确定病毒抗原表达以及结合的免疫球蛋白的抗病毒特异性。使用免疫印迹技术，对这些膜的天然形式以及用高摩尔尿素或非离子去污剂处理后的膜中的WHV核心抗原、e抗原和表面抗原（分别为WHcAg、WHeAg和WHsAg）进行分析。对洗脱物进行检测，以确定是否存在WHV抗原或针对病毒抗原的抗体的反应性。数据显示，急性WHV感染伴随着所检测的所有三种病毒抗原在肝细胞质膜上的表达。在所有情况下，天然膜同时显示WHeAg和WHsAg，而在用尿素或去污剂破坏肝细胞质膜后才检测到WHcAg的存在。数据表明，部分或在某些情况下，甚至全部可检测到的WHcAg特异性可以以一种方式整合到质膜结构中，使得它无法被特异性抗体（抗WHc）识别，这表明在体内条件下，该抗原作为免疫反应靶点至少存在部分功能缺陷。相比之下，在所有研究的天然膜制剂中都可检测到WHeAg特异性，其表达并未明显受到所用处理的影响，而WHsAg的表达则趋于下降。此外，在所有测试的膜洗脱物中都鉴定出了抗WHc反应性，但仅在源自急性肝炎组织学活性处于临界状态且在肝脏灌注前不久血清中e抗原清除的土拨鼠的膜洗脱物中发现了针对WHeAg的抗体（抗WHe）。在洗脱物中未出现针对WHsAg的抗体（抗WHs）。目前的研究结果表明，在急性WHV感染中，WHeAg特异性不仅暴露在受感染肝细胞的表面，而且比WHcAg更容易被血清学识别。上述观察结果表明，除了WHsAg或WHcAg外，e抗原还可作为肝细胞溶解攻击的潜在质膜靶点。此外，本研究结果表明，在急性嗜肝DNA病毒肝炎中，肝脏炎症的低组织学活性、循环中e抗原的清除以及肝细胞质膜结合的抗e抗体的可检测性之间存在明显关系。
