Immunofluorescent detection of bovine papillomavirus E4 antigen in the cytoplasm of cells permissive in vitro for viral DNA amplification.
Immunofluorescent detection of bovine papillomavirus E4 antigen in the cytoplasm of cells permissive in vitro for viral DNA amplification.
复制标题
免疫荧光检测允许体外病毒 DNA 扩增的细胞胞质中的牛乳头瘤病毒 E4 抗原。
DOI：
--
复制
发表时间：
1991
影响因子：
3.8
通讯作者：
S. Burnett
中科院分区：
文献类型：
--
作者：
N. Jareborg;S. Burnett
几种人乳头瘤病毒类型的E4基因在分化的表皮细胞中表达与营养病毒DNA合成相关。为了研究牛乳头瘤病毒1型（BPV-1） E4基因在疣和病毒转化细胞系中的表达，将兔多克隆抗血清与大肠杆菌中作为融合多肽产生的BPV-1 E4抗原联系起来。免疫印迹法检测牛纤维乳头瘤组织中e4相关蛋白16K、21K、30K和42K的表达。在由BPV-1或具有复制能力的BPV-1缺失突变体转化的部分（但不是全部）C127细胞系中，通过免疫荧光分析在静止期培养的细胞亚群中检测到主要定位于核周的细胞质E4抗原。表达e4的细胞通过其粗大的大小来鉴定，以代表与前面显示的相同的细胞亚群，以支持BPV-1 DNA扩增。在该系统中观察到E4蛋白的合成与病毒DNA扩增相关，进一步证明在分裂阻滞细胞亚群中存在病毒早期区域基因表达的开关，这可能准确反映了体内终末分化细胞中BPV-1复制营养期发生的事件。
The E4 gene of several human papillomavirus types is expressed in association with vegetative viral DNA synthesis in differentiated epidermal cells. To develop reagents to study expression of the bovine papillomavirus type 1 (BPV-1) E4 gene in warts and in virus-transformed cell lines, rabbit polyclonal antiserum was raised to the BPV-1 E4 antigen produced as a fusion polypeptide in Escherichia coli. By immunoblotting analysis of productively infected bovine fibropapilloma tissue, E4-related proteins of 16K, 21K, 30K and 42K were detected. In some but not all C127 cell lines transformed by BPV-1 or by a replication-competent BPV-1 deletion mutant, cytoplasmic E4 antigen with a predominantly perinuclear localization was detected by immunofluorescence analysis in a subpopulation of cells in stationary-phase cultures. The E4-expressing cells were identified by their grossly enlarged size to represent the same cell subpopulation shown earlier to support BPV-1 DNA amplification. The observation of synthesis of the E4 protein in association with viral DNA amplification in this system provides further evidence that there is a switch in viral early region gene expression in a subpopulation of division-arrested cells, which may accurately reflect events occurring during the vegetative phase of BPV-1 replication in terminally differentiated cells in vivo.