Huang S N, Millman I, O'Connell A, Aronoff A, Gault H, Blumberg B S
Am J Pathol. 1972 Jun;67(3):453-70.
IN A PREVIOUS REPORT (HUANG SN: Hepatitis-associated antigen hepatitis: an electron microscopic study. Am J Pathol 64:483-500, 1971) liver biopsies of renal transplant patients who developed chronic progressive viral hepatitis associated with persistence of Australia antigenemia [Au(1)] while under immunosuppressive therapy were studied. Predominantly intranuclear 210-250 A spherical, virus-like particles were revealed in 12 of 13 biopsies examined by electron microscope. No such particles were found in biopsies from Australia antigen negative patients. To investigate the relationship of these virus-like particles to Australia antigen, 2 of the Au(1) hepatitis renal transplant patients were restudied 6 to 8 months later. Liver biopsy material was prepared for light microscopy, immunofluorescent microscopy, electron microscopy and immunoelectron microscopy. The specificity of the anti-Au(1) serum used in this study was ascertained by immunodiffusion and immunoelectrophoresis, and by immunoelectron microscopic studies of the antigen-antibody complex prepared in vitro by mixing the ferritin-conjugated anti-Au(1) reagent with the purified Au(1) particles. Electron microscopy of biopsies from liver not treated with antibody showed that virus-like particles persisted in liver cells. Immunofluorescent microscopy of teased liver biopsy suspensions showed nuclear and some cytoplasmic fluorescence, indicating the cellular localization of Au(1). The immunoelectron microscopic preparations showed agglutination of the virus-like particles and the presence of antibody coupled ferritin in the intranuclear and cytoplasmic particle agglutinates. No virus-like particles were seen in the biopsy from a control patient without Au(1) antigenemia, and results of the immunoelectron microscopic procedure were negative. Our observations that massive amounts of Au(1)-associated particles are located in liver cell nuclei of individuals with chronic active hepatitis strengthen the hypothesis of Blumberg et al that Au(1) is an infectious agent and/or the antigenic determinant of a hepatitis virus.
在之前的一份报告中（黄思恩：与肝炎相关抗原性肝炎：一项电子显微镜研究。《美国病理学杂志》64:483 - 500，1971年），对肾移植患者的肝活检组织进行了研究，这些患者在接受免疫抑制治疗期间出现了与澳大利亚抗原血症[Au(1)]持续存在相关的慢性进行性病毒性肝炎。在13份经电子显微镜检查的活检组织中，有12份显示主要为核内210 - 250埃的球形病毒样颗粒。在澳大利亚抗原阴性患者的活检组织中未发现此类颗粒。为了研究这些病毒样颗粒与澳大利亚抗原的关系，对2例Au(1)肝炎肾移植患者在6至8个月后进行了再次研究。制备肝活检材料用于光学显微镜检查、免疫荧光显微镜检查、电子显微镜检查和免疫电子显微镜检查。本研究中使用的抗Au(1)血清的特异性通过免疫扩散和免疫电泳以及通过将铁蛋白偶联抗Au(1)试剂与纯化的Au(1)颗粒在体外混合制备的抗原 - 抗体复合物的免疫电子显微镜研究来确定。未用抗体处理的肝脏活检组织的电子显微镜检查显示病毒样颗粒持续存在于肝细胞中。对肝活检组织悬液进行免疫荧光显微镜检查显示细胞核和一些细胞质有荧光，表明Au(1)的细胞定位。免疫电子显微镜制剂显示病毒样颗粒凝集，并且在核内和细胞质颗粒凝集物中存在抗体偶联的铁蛋白。在无Au(1)抗原血症的对照患者的活检组织中未见到病毒样颗粒，免疫电子显微镜检查结果为阴性。我们观察到大量与Au(1)相关的颗粒位于慢性活动性肝炎患者的肝细胞核中，这强化了布隆伯格等人的假说，即Au(1)是一种感染因子和/或肝炎病毒的抗原决定簇。
