Choi S S, Rasshofer R, Roggendorf M
Max von Pettenkofer-Institute for Hygiene and Medical Microbiology, University of Munich, Federal Republic of Germany.
Virology. 1988 Dec;167(2):451-7.
Monolayer cell cultures of primary woodchuck hepatocytes, prepared by perfusing the liver in situ with collagenase type I, yielded hepatocytes with a viability of greater than 90% which could be held in culture for up to 3 months. Cultures of primary woodchuck hepatocytes were infected one day after plating with hepatitis delta virus (HDV) which had been passaged five times in woodchucks and was therefore identified as woodchuck hepatitis delta virus (WHDV). Replication of WHDV was demonstrated by the appearance of genomic WHDV RNA of ca. 1.7 kb beginning 7 days after infection, with an increase of copy numbers up to 2 weeks after inoculation. Synthesis of hepatitis delta virus-associated antigen (HDAg) in hepatocytes was detected by immunofluorescence staining of hepatocytes. Preincubation of the inoculum with rabbit sera containing antibodies against woodchuck hepatitis virus surface antigen (anti-WHs) reduced the infectivity of WHDV to an undetectable level compared with inocula which were treated with anti-WHs negative sera.
通过用I型胶原酶原位灌注肝脏制备的原代土拨鼠肝细胞单层细胞培养物，所获得的肝细胞活力大于90%，可在培养中维持长达3个月。原代土拨鼠肝细胞培养物在接种一天后用在土拨鼠中传代了5次的丁型肝炎病毒（HDV）进行感染，因此被鉴定为土拨鼠丁型肝炎病毒（WHDV）。感染7天后开始出现约1.7 kb的基因组WHDV RNA，接种后2周内拷贝数增加，证明了WHDV的复制。通过肝细胞免疫荧光染色检测肝细胞中丁型肝炎病毒相关抗原（HDAg）的合成。与用抗WHs阴性血清处理的接种物相比，用含有抗土拨鼠肝炎病毒表面抗原抗体的兔血清预孵育接种物可将WHDV的感染性降低到无法检测的水平。
