Compton T
Department of Medical Microbiology and Immunology, University of Wisconsin-Madison Medical School 53706-1532.
J Virol. 1993 Jun;67(6):3644-8. doi: 10.1128/JVI.67.6.3644-3648.1993.
Human foreskin fibroblasts (HFF) were immortalized via retrovirus-mediated gene transfer of the E6 and E7 genes of human papillomavirus type 16. An immortalized fibroblast (IF) cell line which was morphologically akin to the parental cell line was isolated. The IF cell line was evaluated for permissiveness to human cytomegalovirus (HCMV) infection after the IF cell line surpassed the normal passage limitation of diploid fibroblasts. Western immunoblot analysis of representative HCMV-encoded immediate-early (72-kDa), early (gB), and late (gH) gene products demonstrated that the IF cell line produced these proteins analogous to those produced by the parental HFF cells. Similar quantities of infectious virus were produced in the IF and HFF cell lines as determined in one-step growth curve experiments. Compared with the HFF cells, morphologically identical plaques were produced in the IF cell line in approximately 10 to 12 days postinfection. These findings indicate that fibroblast cell lines immortalized with transforming genes of human papillomavirus retain complete permissiveness to HCMV infection and support plaque formation. The IF cell line will be useful for future genetic analysis of HCMV.
通过逆转录病毒介导的人乳头瘤病毒16型E6和E7基因转移,使人包皮成纤维细胞(HFF)永生化。分离出一种形态与亲代细胞系相似的永生化成纤维细胞(IF)细胞系。在IF细胞系超过二倍体成纤维细胞的正常传代限制后,评估其对人巨细胞病毒(HCMV)感染的易感性。对代表性的HCMV编码的立即早期(72 kDa)、早期(gB)和晚期(gH)基因产物进行的蛋白质免疫印迹分析表明,IF细胞系产生这些蛋白质的情况与亲代HFF细胞产生的情况类似。在一步生长曲线实验中确定,IF和HFF细胞系产生的感染性病毒量相似。与HFF细胞相比,在感染后约10至12天,IF细胞系中产生了形态相同的噬斑。这些发现表明,用人乳头瘤病毒转化基因永生化的成纤维细胞系对HCMV感染保持完全易感性并支持噬斑形成。IF细胞系将有助于未来对HCMV进行基因分析。