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传染性软疣病毒在原代人成纤维细胞中表达晚期基因,但不产生有感染性的子代病毒。

Molluscum contagiosum virus expresses late genes in primary human fibroblasts but does not produce infectious progeny.

作者信息

Bugert J J, Melquiot N, Kehm R

机构信息

Department of Virology, Ruprecht-Karls-Universität Heidelberg.

出版信息

Virus Genes. 2001 Jan;22(1):27-33. doi: 10.1023/a:1008126217725.

Abstract

Molluscum contagiosum virus (MCV), a member of the family Poxviridae, can be isolated from skin-lesion material of patients with molluscum contagiosum infection. MCV replicates efficiently in human keratinocytes in vivo but viral replication has not been observed in vitro in cell or tissue culture systems. We investigated a variety of established cell lines for productive MCV infection and found that: (i) MCV induces a typical cytopathogenic effect (CPE) only in human primary fibroblast cells (MRC5 ATCC-CCL 171 and HEPM ATCC-CRL 1486) but not in permanent eucaryotic cell lines of human or simian origin; (ii) UV irradiated MCV virions and heat inactivated virions do not induce a CPE; (iii) decreasing amounts of MCV viral DNA are detectable in infected human embryonic fibroblasts for at least 14 days post infection (p.i.); (iv) MCV early mRNAs are detectable by RT-PCR between one and two hours p.i. and remain detectable upon passaging of the infected cells; (iv) transcripts of viral late genes (mc095L and mc106L) are detectable by RT-PCR from day 5 p.i.; (v) MCV viral antigens can be detected on the surface of infected cells using human and rabbit polyclonal antisera against MCV from 24 h p.i.; (vi) a CPE can not be observed if cell free supernatants or homogenizates of MCV infected cells are used to try passage of the virus onto uninfected human embryonic fibroblasts, indicating that infectious viral progeny is not produced. This is the first report demonstrating long time persistence of MCV viral DNA and expression of late proteins in an in vitro cell culture system.

摘要

传染性软疣病毒(MCV)是痘病毒科的成员,可从传染性软疣感染患者的皮肤病变材料中分离出来。MCV在体内的人角质形成细胞中能高效复制,但在细胞或组织培养系统中尚未观察到其在体外的病毒复制。我们研究了多种已建立的细胞系以检测MCV的有效感染情况,结果发现:(i)MCV仅在人原代成纤维细胞(MRC5 ATCC-CCL 171和HEPM ATCC-CRL 1486)中诱导典型的细胞病变效应(CPE),而在人源或猿源的永生化真核细胞系中则不会;(ii)紫外线照射的MCV病毒粒子和热灭活的病毒粒子不会诱导CPE;(iii)在感染后的至少14天内,在感染的人胚胎成纤维细胞中可检测到数量逐渐减少的MCV病毒DNA;(iv)在感染后1至2小时之间通过RT-PCR可检测到MCV早期mRNA,并且在感染细胞传代后仍可检测到;(iv)从感染后第5天起通过RT-PCR可检测到病毒晚期基因(mc095L和mc106L)的转录本;(v)从感染后24小时起,使用抗MCV的人源和兔源多克隆抗血清可在感染细胞表面检测到MCV病毒抗原;(vi)如果使用MCV感染细胞的无细胞上清液或匀浆来尝试将病毒传代至未感染的人胚胎成纤维细胞上,则不会观察到CPE,这表明未产生有传染性的病毒后代。这是第一份证明MCV病毒DNA在体外细胞培养系统中长时间持续存在以及晚期蛋白表达的报告。

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