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建立持续感染原型BK多瘤病毒的COS-BK细胞。

Establishment of COS-BK cells persistently infected with archetype BK polyomavirus.

作者信息

Nukuzuma Souichi, Onogi Hiroshi, Suzuki Tetsuro

机构信息

Research Laboratory, KinoPharma Inc., Kyoto, Japan.

Department of Microbiology and Immunology, Hamamatsu University School of Medicine, Hamamatsu, Shizuoka, Japan.

出版信息

Microbiol Immunol. 2024 May;68(5):179-184. doi: 10.1111/1348-0421.13124. Epub 2024 Mar 3.

Abstract

BK polyomavirus (BKPyV) was the first human polyomavirus to be isolated from an immunosuppressed kidney transplant recipient in 1971. BKPyV reactivation causes BKPyV-associated nephropathy and hemorrhagic cystitis. However, the mechanisms underlying BKPyV replication remain unclear. In the present study, we performed the long-term cultivation of COS-7 cells transfected with archetype KOM-5 DNA, which were designated as COS-BK cells. BKPyV derived from COS-BK cells was characterized by analyzing the amount of the virus based on hemagglutination, viral replication, and the production of viral protein 1 (VP1). Immunostaining showed that VP1-positive cells accounted for a small percentage of COS-BK cells. The nucleotide sequences encompassing the origin of the DNA replication of BKPyV derived from COS-BK cells were generated from KOM-5 by the deletion of an 8-bp sequence, which did not involve T antigen binding sites. BKPyV replicated most efficiently in COS-BK cells in DMEM containing 2% fetal bovine serum. These results indicate that COS-BK cells are a suitable culture system for studying the persistent infection of archetype BKPyV.

摘要

BK多瘤病毒(BKPyV)是1971年从一名免疫抑制的肾移植受者中分离出的首例人类多瘤病毒。BKPyV再激活会导致BKPyV相关性肾病和出血性膀胱炎。然而,BKPyV复制的潜在机制仍不清楚。在本研究中,我们对转染了原型KOM-5 DNA的COS-7细胞进行了长期培养,这些细胞被命名为COS-BK细胞。通过基于血凝、病毒复制和病毒蛋白1(VP1)产生来分析病毒量,对源自COS-BK细胞的BKPyV进行了表征。免疫染色显示,VP1阳性细胞在COS-BK细胞中占比很小。通过缺失一个8碱基对序列(该序列不涉及T抗原结合位点)从KOM-5生成了包含源自COS-BK细胞的BKPyV DNA复制起点的核苷酸序列。BKPyV在含有2%胎牛血清的DMEM中在COS-BK细胞中复制效率最高。这些结果表明,COS-BK细胞是研究原型BKPyV持续感染的合适培养系统。

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