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人细小病毒B19在源自胎儿肝脏的红系谱系细胞原代培养中的增殖。

Propagation of human parvovirus B19 in primary culture of erythroid lineage cells derived from fetal liver.

作者信息

Yaegashi N, Shiraishi H, Takeshita T, Nakamura M, Yajima A, Sugamura K

机构信息

Department of Bacteriology, Tohoku University School of Medicine, Sendai, Japan.

出版信息

J Virol. 1989 Jun;63(6):2422-6. doi: 10.1128/JVI.63.6.2422-2426.1989.

Abstract

Erythroid lineage cells derived from fetal liver were demonstrated to be target cells for human parvovirus B19 infection. B19 virus antigen-positive serum was inoculated into primary cultures containing erythroid lineage cells enriched from fetal liver. The B19 virus antigen was detected on about 5% of cells in the culture by immunofluorescence staining, and the stained cells were identified as erythroid lineage cells by double staining with anti-B19 virus-positive serum and anti-erythroid lineage monoclonal antibody. The immunofluorescence staining study also revealed that the B19 virus antigen localized in the nucleus and the periphery of cytoplasm. We also detected B19 virus DNA, which was generated by replication in the infected cells, not only in the cells but also in the culture supernatants, in which the amount of B19 DNA increased depending on the period of culture, indicating that the cells infected with B19 virus produced B19 virus and released it into the medium. The ability of B19 virus released into the medium to infect fetal erythroid lineage cells was demonstrated quantitatively. Because of the absence of any cytopathic effect of B19 virus during culture periods of at least 15 days, this culture system should be useful in the study of B19 virus replication and in vitro generation of B19 virus. In addition, the present study may contribute to a better understanding of the pathogenesis of hydrops fetalis, which is probably associated with B19 virus infection during pregnancy.

摘要

源自胎儿肝脏的红系细胞被证明是人细小病毒B19感染的靶细胞。将B19病毒抗原阳性血清接种到含有从胎儿肝脏富集的红系细胞的原代培养物中。通过免疫荧光染色在培养物中约5%的细胞上检测到B19病毒抗原,并且通过用抗B19病毒阳性血清和抗红系单克隆抗体进行双重染色,将染色的细胞鉴定为红系细胞。免疫荧光染色研究还显示B19病毒抗原定位于细胞核和细胞质周边。我们还检测到B19病毒DNA,其在感染细胞中通过复制产生,不仅存在于细胞中,还存在于培养上清液中,其中B19 DNA的量根据培养时间而增加,这表明感染B19病毒的细胞产生了B19病毒并将其释放到培养基中。定量证明了释放到培养基中的B19病毒感染胎儿红系细胞的能力。由于在至少15天的培养期间B19病毒没有任何细胞病变效应,该培养系统应有助于研究B19病毒复制和B19病毒的体外产生。此外,本研究可能有助于更好地理解胎儿水肿的发病机制,胎儿水肿可能与孕期B19病毒感染有关。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1b7c/250692/5ea8582ddb25/jvirol00073-0022-a.jpg

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