Galle P R, Schlicht H J, Kuhn C, Schaller H
Zentrum für Molekulare Biologie Heidelberg, Federal Republic of Germany.
Hepatology. 1989 Oct;10(4):459-65. doi: 10.1002/hep.1840100410.
Primary duck hepatocytes obtained from Pekin ducks congenitally infected with duck hepatitis B virus were used to monitor expression of viral proteins and replication of viral DNA in cell culture. Duck hepatitis B virus core antigen, duck hepatitis B virus pre-surface antigen and duck hepatitis B virus DNA were detectable for at least 12 days after cell plating. Whereas expression of duck hepatitis B pre-surface antigen was constant during this time, expression of duck hepatitis B core antigen and of viral DNA rapidly declined. This diminished production of viral components in vitro was paralleled by a change of the hepatocytes toward a fibroblast-like morphology. Supplementation of cell culture medium with 2% dimethyl sulfoxide, a solvent known to maintain the differentiated state of cultured cells, retained competence of the cultured hepatocytes to express duck hepatitis B core antigen and duck hepatitis B virus DNA at high levels. In a second set of experiments, duck hepatitis B virus negative hepatocytes were infected with duck hepatitis B virus from serum of congenitally infected ducks. Dimethyl sulfoxide remarkably improved the competence of cultured duck hepatocytes to become productively infected. This function was maintained for at least 12 days postplating.
从先天性感染鸭乙型肝炎病毒的北京鸭中获取的原代鸭肝细胞,用于监测细胞培养中病毒蛋白的表达和病毒DNA的复制。在细胞接种后至少12天内可检测到鸭乙型肝炎病毒核心抗原、鸭乙型肝炎病毒前表面抗原和鸭乙型肝炎病毒DNA。在此期间,鸭乙型肝炎前表面抗原的表达保持恒定,而鸭乙型肝炎核心抗原和病毒DNA的表达迅速下降。体外病毒成分产量的减少与肝细胞向成纤维细胞样形态的转变同时发生。向细胞培养基中添加2%的二甲基亚砜(一种已知可维持培养细胞分化状态的溶剂),可使培养的肝细胞保持高水平表达鸭乙型肝炎核心抗原和鸭乙型肝炎病毒DNA的能力。在另一组实验中,用先天性感染鸭血清中的鸭乙型肝炎病毒感染鸭乙型肝炎病毒阴性肝细胞。二甲基亚砜显著提高了培养的鸭肝细胞发生有效感染的能力。该功能在接种后至少12天内得以维持。