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

IF 1.9 4区 医学 Q4 IMMUNOLOGY Microbiology and Immunology Pub Date : 2024-03-03 DOI:10.1111/1348-0421.13124
Souichi Nukuzuma, Hiroshi Onogi, Tetsuro Suzuki
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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.

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建立持续感染原型 BK 多瘤病毒的 COS-BK 细胞。
BK 多瘤病毒(BKPyV)是 1971 年从免疫抑制肾移植受者体内分离出的第一种人类多瘤病毒。BKPyV 再活化会导致 BKPyV 相关性肾病和出血性膀胱炎。然而,BKPyV 的复制机制仍不清楚。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,并将其命名为 COS-BK 细胞。我们根据血凝、病毒复制和病毒蛋白 1(VP1)的产生情况分析了病毒的数量,从而确定了从 COS-BK 细胞中提取的 BKPyV 的特征。免疫染色显示,VP1 阳性细胞只占 COS-BK 细胞的一小部分。从 COS-BK 细胞中提取的 BKPyV DNA 复制源的核苷酸序列是从 KOM-5 中删除 8-bp 序列后产生的,该序列不涉及 T 抗原结合位点。在含有 2% 胎牛血清的 DMEM 中,BKPyV 在 COS-BK 细胞中的复制效率最高。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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来源期刊
Microbiology and Immunology
Microbiology and Immunology 医学-免疫学
CiteScore
5.20
自引率
3.80%
发文量
78
审稿时长
1 months
期刊介绍: Microbiology and Immunology is published in association with Japanese Society for Bacteriology, Japanese Society for Virology, and Japanese Society for Host Defense Research. It is peer-reviewed publication that provides insight into the study of microbes and the host immune, biological and physiological responses. Fields covered by Microbiology and Immunology include:Bacteriology|Virology|Immunology|pathogenic infections in human, animals and plants|pathogenicity and virulence factors such as microbial toxins and cell-surface components|factors involved in host defense, inflammation, development of vaccines|antimicrobial agents and drug resistance of microbes|genomics and proteomics.
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