AMPLIFICATION OF HUMAN PAPILLOMAVIRUS GENOMES INVITRO IS DEPENDENT ON EPITHELIAL DIFFERENTIATION
AMPLIFICATION OF HUMAN PAPILLOMAVIRUS GENOMES INVITRO IS DEPENDENT ON EPITHELIAL DIFFERENTIATION
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DOI：
10.1128/jvi.65.5.2254-2260.1991
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发表时间：
1991-05-01
影响因子：
5.4
通讯作者：
LAIMINS, LA
中科院分区：
文献类型：
--
作者：
BEDELL, MA;HUDSON, JB;LAIMINS, LA
人乳头瘤病毒(HPV)感染鳞状上皮，并在增殖的基底细胞中建立其基因组的上皮体。随着感染细胞的分化，病毒DNA被放大到高拷贝数，从而产生传染性病毒。由于标准培养方法不能复制上皮细胞分化的大部分阶段，在体外还没有观察到病毒的产生。在这项研究中，我们检查了一个来自低级别宫颈病变的细胞系，发现它含有HPV-31亚型HPV-31b的外体拷贝，每个细胞大约有50个拷贝。当允许在体外RAFT培养的气液界面分层时，该细胞系以一种组织学上类似于体内低级别宫颈病变的方式分化。我们已经观察到HPV-31b基因组在体外复层上皮上部的不同部位的扩增，类似于体内产生性HPV感染。虽然在分层RAFT培养中也检测到HPV-31b晚期区域的转录本，但没有发现衣壳蛋白。这些研究复制了体外高效HPV感染的一个重要方面，即依赖于分化的乳头瘤病毒基因组的扩增。
Human papillomaviruses (HPVs) infect squamous epithelium and establish their genomes as episomes in proliferating basal cells. As infected cells differentiate, the viral DNA is amplified to high copy number and infectious virus is produced. Viral production has not yet been observed in vitro due to the inability of standard culture methods to duplicate most stages of epithelial differentiation. In this study, we have examined a cell line derived from a low-grade cervical lesion and found that it contained episomal copies of an HPV-31 subtype, HPV-31b, at approximately 50 copies per cell. When allowed to stratify at the air-liquid interface of in vitro raft cultures, this cell line differentiates in a manner which histologically resembles a low-grade cervical lesion in vivo. We have observed the amplification of HPV-31b genomes in distinct foci in the upper portion of the in vitro-stratified epithelium similar to that found in productive HPV infections in vivo. Although transcripts from the late region of HPV-31b were also detected specifically in stratified raft cultures, no capsid protein was found. These studies duplicate one important aspect of a productive HPV infection in vitro, the differentiation-dependent amplification of papillomavirus genomes.