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Propagation of human hepatitis A virus in African green monkey kidney cell culture: primary isolation and serial passage.

机译:人类甲型肝炎病毒在非洲绿猴肾细胞培养物中的传播:初步分离和连续传代。

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摘要

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.
机译:人类甲型肝炎病毒(HAV)在原发非洲绿猴(Cercopithecus aethiops)肾脏(AGMK)细胞培养物中繁殖。使用了三种HAV菌株:MS-1,SD-11和HM-175。用mar猴传代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确定病毒的身份。临床样品和mar猴传代的材料均产生免疫荧光。发现HAV抗原散布在接种培养物的整个细胞质中。 HM-175菌株产生最强烈的免疫荧光。该HAV株已在细胞培养物中连续传代7次。用来自自然感染的HAV感染的成对人类血清和来自实验感染的动物的超免疫黑猩猩血清进行的阻断实验证实,免疫荧光是特异性的。发现病毒抗原仅在细胞内。通过连续传代,最大HAV抗原表达的间隔减小。直接从原代AGMK细胞培养中的HAV患者粪便样本中回收的本文所述的HAV株可证明可用作血清学测试的抗原来源和候选疫苗株。

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