Fagan E, Yousef G, Brahm J, Garelick H, Mann G, Wolstenholme A, Portmann B, Harrison T, Mowbray J F, Mowat A
Liver Unit, King's College Hospital, London, England.
J Med Virol. 1990 Feb;30(2):131-6. doi: 10.1002/jmv.1890300210.
A peroxidase-labelled, specific mouse monoclonal antibody to hepatitis A virus (HAV) and an in situ hybridization technique (streptavidin-biotin-horseradish peroxidase reaction) with an HAV-specific cDNA probe (recombinant plasmid pAWHA comprising 1.8 kb of the HAV-specific cDNA, located toward the 3' end of the genome) were used to detect HAV in liver tissues in two patients with fulminant viral hepatitis type A treated by liver transplantation after a protracted (day 40: case 1) and relapsing (day 60: case 2) clinical course. HAV antigens and HAV-specific genomic sequences were detected in the hepatectomy tissues and in serial biopsies of the liver grafts through to final follow-up at 2 months (case 2) or death at 7 months after re-grafting for chronic rejection (case 1). In the fulminant liver parenchyma, numerous degenerating and some surviving hepatocytes were positive and randomly scattered. The immunoperoxidase staining was predominantly cytoplasmic and often granular. The localization of the cDNA probe was predominantly nuclear/perinuclear but was occasionally cytoplasmic. High-titre IgM-anti-HAV antibodies persisted until death (case 1) or resolution (5 months) of an acute hepatitis (case 2), which occurred at 2 months, accompanied by HAV antigen (ELISA), in stool. Intact replicating virus particles must have been present in one or more sites in each case, including extrahepatic locations, with a viraemia as the most likely explanation for subsequent reinfection of the grafts.
使用一种过氧化物酶标记的、针对甲型肝炎病毒（HAV）的特异性小鼠单克隆抗体以及一种原位杂交技术（链霉亲和素-生物素-辣根过氧化物酶反应），结合HAV特异性cDNA探针（重组质粒pAWHA，包含1.8 kb的HAV特异性cDNA，位于基因组的3'端），对两名接受肝移植治疗的暴发性甲型病毒性肝炎患者的肝组织进行HAV检测。这两名患者分别经历了迁延性（第40天：病例1）和复发性（第60天：病例2）临床病程。在肝切除组织以及肝移植的系列活检组织中均检测到了HAV抗原和HAV特异性基因组序列，直至最终随访，病例2随访至2个月，病例1因慢性排斥反应在再次移植后7个月死亡。在暴发性肝炎的肝实质中，许多变性的肝细胞以及一些存活的肝细胞呈阳性，且随机分布。免疫过氧化物酶染色主要位于细胞质，且常呈颗粒状。cDNA探针的定位主要在细胞核/核周，但偶尔也在细胞质。高滴度的IgM抗HAV抗体一直持续到死亡（病例1）或急性肝炎消退（5个月，病例2），病例2在2个月时急性肝炎消退，同时粪便中检测到HAV抗原（ELISA法）。在每个病例中，完整的复制病毒颗粒必定存在于一个或多个部位，包括肝外部位，病毒血症很可能是随后移植肝再次感染的原因。
