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目的:研究证实HIV-1感染相关细胞因子能够诱导原发性渗出性淋巴瘤(PEL)的另一细胞系BC-3中潜伏的人类疱疹病毒8型(HHV-8)发生可溶性周期复制。方法:将TNF-α和HIV-1感染CD4~+ T淋巴细胞常释放的细胞因子IFN-γ、HGF/SF、OSM,连续加入到BC-3细胞中作持续刺激,分别于刺激后的第3天和第7天收集BC-3细胞。采用免疫组化染色法(IHC)检测HHV-8免疫原性蛋白ORF59表达;电子显微镜观察病毒的形成;Northernblot和 定量PCR检查次要衣壳蛋白ORF26 mRNA转录。结果:IFN-γ、HGF/SF、OSM和TNF-α均能不同程度上调ORF26 mRNA表达。其中,IFN-γ刺激BC-3细胞第7天,HHV-8 ORF26 mRNA表达上升了6.1倍;ORF59蛋白表达上升到20％,与此同时,BC-3细胞中可观察到成熟的病毒粒子。另外,TNF-α刺激BC-3细胞第7天,HHV-8 ORF26 mRNA表达也上升了2.5。结论:TNF-α和HIV-1感染释放细胞因子能够诱导PEL另一细胞系BC-3中潜伏的HHV-8发生可溶性周期复制。
PURPOSE: To demonstrate soluble-cycle replication of latent human herpesvirus type 8 (HHV-8) in another cell line, BC-3, that demonstrates that HIV-1-associated cytokines can induce primary exudative lymphoma (PEL). Methods: The cytokines such as IFN-γ, HGF / SF and OSM released by TNF-α and HIV-1 infected CD4 ~ + T lymphocytes were continuously added to BC-3 cells for continuous stimulation. BC-3 cells were collected on days 3 and 7. Immunohistochemical staining (IHC) was used to detect the expression of HHV-8 immunogenic protein ORF59; electron microscopy was used to observe the formation of the virus; Northern blots and quantitative PCR were used to examine the ORF26 mRNA transcription of the minor capsid protein. Results: IFN-γ, HGF / SF, OSM and TNF-α all up-regulated ORF26 mRNA expression to some extent. Among them, the expression of HHV-8 ORF26 mRNA increased by 6.1-fold on the 7th day in BC-3 cells stimulated by IFN-γ, and the expression of ORF59 protein increased to 20%. At the same time, mature virus particles were observed in BC-3 cells. In addition, on day 7 of BC-3 cells stimulated by TNF-α, the expression of HHV-8 ORF26 mRNA also increased by 2.5. CONCLUSION: The release of cytokines by TNF-α and HIV-1 infection induced soluble cyclic replication of latent HHV-8 in BC-3, another cell line of PEL.