Heinricy U, Stierhof Y D, Pfisterer M, Flehmig B
Department of Medical Virology and Epidemiology of Viral Diseases, University of Tuebingen, F.R.G.
J Gen Virol. 1987 Sep;68 ( Pt 9):2487-93. doi: 10.1099/0022-1317-68-9-2487.
This paper describes the biophysical and biochemical properties as well as electron microscopical studies of a candidate hepatitis A vaccine strain propagated in human fibroblast cells. Our results indicated that, in CsCl, the density of hepatitis A virus (HAV) from cell culture supernatant and of HAV extracted from infected cells was influenced by the quantity of lipid material associated with HAV. Antigenicity of untreated HAV, therefore, was detected primarily in low density CsCl fractions (1.11 g/ml, 1.21 g/ml). After lipid reduction with NP40 detergent or chloroform/Genetron, antigenicity and infectivity were primarily detected in high density CsCl fractions (1.31 g/ml). Electron microscopy demonstrated a strong association between membranous material and virus particles of low density in CsCl as well as virus-like particles in ultrathin sections of HAV-infected human fibroblast cells. The uncovered virus particles banded in the 1.31 g/ml dense CsCl fraction lacked lipid material. The s value was 79 for 1.19 g/ml to 1.22 g/ml dense HAV and 147 for 1.29 g/ml to 1.33 g/ml HAV. Autoradiography of the radioiodinated dense HAV revealed some proteins with high Mr (120K to 67K) and others with low Mr (37K to 15K).
本文描述了在人成纤维细胞中增殖的甲型肝炎疫苗候选毒株的生物物理和生化特性以及电子显微镜研究。我们的结果表明，在氯化铯中，细胞培养上清液中的甲型肝炎病毒（HAV）以及从感染细胞中提取的HAV的密度受与HAV相关的脂质物质数量的影响。因此，未处理的HAV的抗原性主要在低密度氯化铯组分（1.11 g/ml，1.21 g/ml）中检测到。用NP40去污剂或氯仿/氟利昂进行脂质去除后，抗原性和感染性主要在高密度氯化铯组分（1.31 g/ml）中检测到。电子显微镜显示，在氯化铯中的低密度病毒颗粒以及HAV感染的人成纤维细胞超薄切片中的病毒样颗粒与膜状物质之间有很强的关联。在1.31 g/ml密度的氯化铯组分中形成条带的裸露病毒颗粒缺乏脂质物质。密度为1.19 g/ml至1.22 g/ml的HAV的s值为79，密度为1.29 g/ml至1.33 g/ml的HAV的s值为147。放射性碘化高密度HAV的放射自显影显示出一些高分子量（120K至67K）的蛋白质和其他低分子量（37K至15K）的蛋白质。
