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根据细胞周期状态增强对细小病毒 B19V 感染单位的基于细胞的检测。

Enhanced Cell-Based Detection of Parvovirus B19V Infectious Units According to Cell Cycle Status.

机构信息

Laboratoire Français du Fractionnement et des Biotechnologies (LFB), 3 avenue des Tropiques, Les Ulis, BP 305, 91958 Courtabœuf Cedex, France.

Division of Innovative Therapies, UMR-1184, IMVA-HB and IDMIT Center, CEA, INSERM and Paris-Saclay University, F-92265 Fontenay-aux-Roses, France.

出版信息

Viruses. 2020 Dec 18;12(12):1467. doi: 10.3390/v12121467.

DOI:10.3390/v12121467
PMID:33353185
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7766612/
Abstract

Human parvovirus B19 (B19V) causes various human diseases, ranging from childhood benign infection to arthropathies, severe anemia and fetal hydrops, depending on the health state and hematological status of the patient. To counteract B19V blood-borne contamination, evaluation of B19 DNA in plasma pools and viral inactivation/removal steps are performed, but nucleic acid testing does not correctly reflect B19V infectivity. There is currently no appropriate cellular model for detection of infectious units of B19V. We describe here an improved cell-based method for detecting B19V infectious units by evaluating its host transcription. We evaluated the ability of various cell lines to support B19V infection. Of all tested, UT7/Epo cell line, UT7/Epo-STI, showed the greatest sensitivity to B19 infection combined with ease of performance. We generated stable clones by limiting dilution on the UT7/Epo-STI cell line with graduated permissiveness for B19V and demonstrated a direct correlation between infectivity and S/G2/M cell cycle stage. Two of the clones tested, B12 and E2, reached sensitivity levels higher than those of UT7/Epo-S1 and CD36 erythroid progenitor cells. These findings highlight the importance of cell cycle status for sensitivity to B19V, and we propose a promising new straightforward cell-based method for quantifying B19V infectious units.

摘要

人细小病毒 B19（B19V）可引起多种人类疾病，范围从儿童良性感染到关节病、严重贫血和胎儿水肿，这取决于患者的健康状况和血液状况。为了对抗 B19V 的血液污染，对血浆池中的 B19 DNA 进行评估，并采取病毒灭活/去除步骤，但核酸检测不能正确反映 B19V 的感染力。目前，尚无用于检测 B19V 感染性单位的适当细胞模型。我们在这里描述了一种改进的基于细胞的方法，通过评估其宿主转录来检测 B19V 感染性单位。我们评估了各种细胞系支持 B19V 感染的能力。在所有测试的细胞系中，UT7/Epo 细胞系和 UT7/Epo-STI 细胞系对 B19 感染的敏感性最高，并且易于操作。我们在 UT7/Epo-STI 细胞系上通过有限稀释生成了具有 B19V 逐渐许可性的稳定克隆，并证明了感染性与 S/G2/M 细胞周期阶段之间存在直接相关性。测试的两个克隆，B12 和 E2，其敏感性水平高于 UT7/Epo-S1 和 CD36 红系祖细胞。这些发现强调了细胞周期状态对 B19V 敏感性的重要性，我们提出了一种有前途的新的简单基于细胞的方法来定量 B19V 感染性单位。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1eff/7766612/b0cd9f440402/viruses-12-01467-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1eff/7766612/7caf91697b1a/viruses-12-01467-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1eff/7766612/874d20a2ffc7/viruses-12-01467-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1eff/7766612/89da7a663c14/viruses-12-01467-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1eff/7766612/0fb1c19d6c4b/viruses-12-01467-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1eff/7766612/33890c7c09e8/viruses-12-01467-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1eff/7766612/a372990858dc/viruses-12-01467-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1eff/7766612/cce27bd25b0a/viruses-12-01467-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1eff/7766612/b0cd9f440402/viruses-12-01467-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1eff/7766612/7caf91697b1a/viruses-12-01467-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1eff/7766612/874d20a2ffc7/viruses-12-01467-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1eff/7766612/89da7a663c14/viruses-12-01467-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1eff/7766612/0fb1c19d6c4b/viruses-12-01467-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1eff/7766612/33890c7c09e8/viruses-12-01467-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1eff/7766612/a372990858dc/viruses-12-01467-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1eff/7766612/cce27bd25b0a/viruses-12-01467-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1eff/7766612/b0cd9f440402/viruses-12-01467-g008.jpg

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