卡波济肉瘤相关疱疹病毒体外感染人牙龈成纤维细胞初探

摘要

Objective: To establish a model of infecting human gingival fibroblast(HGF)cells with KSHVin vitro for further studying the mechanism of oral KS pathogenesy. Methods:The supernatants containing KSHV virion harvested from primary effusion lymphoma (PEL) cells BC-3 were used to infect HGF cells. The cytopathic effect (CPE) of HGF cells was observed post-infection and the lyric replication of KSHV in HGF cells was detected by RT-PCR and Western blot. Results:CPE appeared in post-infective HGF cells. KSHV ORF26 and ORF73 mRNA transcription was detected in HGF cells by RT-PCR at diffenrent time points. And the specific band of vIL-6 protein was detected at 12 hours post-infection by Western blot Conclusion:KSHV could infect HGF cells and establish latent infection,providing a modelin vitro for further studying the mechanism in KSHV leading to oral KS.%目的:探索卡波济肉瘤相关疱疹病毒(Kaposi's sarcoma-associated herpesvirus,KSHV)体外感染人牙龈成纤维细胞(human gingival fibroblast,HGF)的途径和方法,为研究KSHV导致口腔卡波济肉瘤病变的机制提供依据.方法:用12-O-十四烷酰佛波醇-13-乙酯(12-O-tetradecanoylphorbol-13-acetate,TPA)刺激人原发性渗出性淋巴瘤(primary effusion lymphoma,PEL)细胞系的BC-3细胞,收集细胞上清(含KSHV病毒颗粒),感染HGF细胞.观察细胞病变效应(cytopatric effect,CPE).采用RT-PCR和Western blot分别检测HGF细胞内KSHV编码基因的转录情况和蛋白表达水平.结果:KSHV感染HGF细胞后可出现CPE;感染后采用RT-PCR可检测到不同时间点ORF26和ORF73基因的转录;感染后12 h可检测到vIL-6蛋白的表达.结论:KSHV可感染HGF细胞并建立潜伏感染,为进一步研究KSHV导致的口腔kS发病机制提供了较好的体外模型.