甲肝病毒在人血白细胞中体外连续传代和在分类白细胞中增殖的研究

Experimental Studies on Serial Passage of HAV in Human Leukocytes and Propagation of HAV In Various Kinds of Leukocytes in Vitro

1985年我们采用间接免疫荧光法(IF法)检测出甲肝患者外周血白细胞中有甲肝抗原(HAAg)存在,继而又将HAAg阳性白细胞直接种入PLC/PRF/5细胞,分离到两株甲肝病毒(HAV)NJ—3株和H—1株。为了弄清白细胞所携带的病毒究竟仅为吸附吞饮,抑或能在其中复制增殖,我们将分离到的HAV用正常人血白细胞进行体外增殖试验,现将结果报告如下。

This paper reports the experimental studies on propagation of HAV in the human mononuclear leukocytes in vitro. The mononuelear leukocytes were separated from the peripheral blood of health person and the umbilical blood of fetus using Ficoll-Hypaque technique. The NJ-3 strain of HAV was inoclated into the adherent cells in culture flask and the suspensions of mononuclear leukocytes. After incubation the cultures were examined by indirect IF technique. It was found that both the adherent and nonadherent mononuclear leukocytes could be infected and propagated by HAV The numbers of leukocytes with IF granules of HAV were found to be increased as times went on and also with the increasing passages. On serial passage HAV could propagate in the leukocytes over 4 passages TCID_(50) increased from 10^(-1)/ml at passage 2 to 10^(-4.25)/ml at passage 4. In further studies, the monoeytes and T or B lymphoeytes were separated and purified using E-Rosette Forming method. The NJ-3 strain of HAV was inoculated into the cells respectively. After ineubation, the results indicated that HAV could propagate in monoeytes and T cells but hardly or not propagate in B lymphocytes.