Daemer R J, Feinstone S M, Gust I D, Purcell R H
Infect Immun. 1981 Apr;32(1):388-93. doi: 10.1128/iai.32.1.388-393.1981.
Human hepatitis A virus (HAV) was propagated in primary African Green Monkey (Cercopithecus aethiops) kidney (AGMK) cell cultures. Three strains of HAV were used: MS-1, SD-11, and HM-175. Cells were inoculated with marmoset-passaged material or human clinical specimens and were stained by direct immunofluorescence to establish the identity of the virus. Both clinical samples and marmoset-passaged material produced immunofluorescence. HAV antigen was found scattered throughout the cytoplasm of inoculated cultures. The HM-175 strain produced the most intense immunofluorescence. This strain of HAV had been serially passaged in cell culture seven times. Blocking experiments with paired human sera from naturally acquired HAV infections and hyperimmune chimpanzee serum from an experimentally infected animal established that the immunofluorescence was specific. The viral antigen was found to be exclusively intracellular. The interval to maximum HAV antigen expression was decreased by serial passage. The HAV strain described herein, which was recovered directly from the stool specimen of a patient with HAV in primary AGMK cell culture, may prove useful as a source of antigen for serological tests and as a candidate vaccine strain.
人甲型肝炎病毒(HAV)在原代非洲绿猴(猕猴)肾(AGMK)细胞培养物中增殖。使用了三株HAV:MS-1、SD-11和HM-175。用传代于狨猴的材料或人类临床标本接种细胞,并通过直接免疫荧光染色来确定病毒的身份。临床样本和传代于狨猴的材料均产生免疫荧光。在接种的培养物中发现HAV抗原散布于整个细胞质中。HM-175毒株产生的免疫荧光最强。该毒株的HAV已在细胞培养中连续传代7次。用来自自然感染HAV的成对人血清和来自实验感染动物的超免疫黑猩猩血清进行的阻断实验证实,这种免疫荧光是特异性的。发现病毒抗原仅存在于细胞内。连续传代可缩短达到最大HAV抗原表达的间隔时间。本文所述的HAV毒株是直接从一名甲型肝炎患者的粪便标本中在原代AGMK细胞培养物中分离得到的,可能作为血清学检测的抗原来源和候选疫苗毒株有用。