HIV-1感染相关细胞因子激活BC-3细胞中潜伏感染的人类疱疹病毒8型的复制研究

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。

Reactivation of Latent Infection of Human Herpesvirus 8 in BC-3 Cells from Primary Effusion Lymphoma by Recombinant Cytokines Similar to that Produced by HIV-1-infected T Cells

Objective: To study and confirm that recombinant cytokines similar to those produced by HIV-1 infected T cells induced lytic cycle replication of human herpesvirus 8 (HHV-8) in BC-3 cells, another cell line from primary effusion lymphoma (PEL). Methods: The persistent stimulation of BC-3 was conducted by several cytokines known to be produced by HIV-1-infected T cells and important in growth and proliferation of Kaposi's sarcoma(KS)cells in vitro, such as the interferonγ (IFN-γ), the hepatoeyte growth factor/scatter factor (HGF/SF), the Oncostain M (OSM), and the tumor necrosis factor-a (TNF-a)which is not produced by HIV-1-infected T cells. Treated and un treated BC-3 cells were collected at the 3rd and 7th day of persistent stimulation, respectively. Immunohistochemical (IHC) staining, Northern blot, quantitative PCR (real-time PCR) and electron microscopy (EM) were carried out to detect the expresswn of immunogenic protein ORF59, messenger RNA (mRNA) of minor capsid protein ORF26, and the presence of viral particles of HHV-8 from treated and untreated BC-3 cells. Results: It showed that IFN-γ, HGF/SF, OSM, and TNF-a were found to induce an increase in mRNA expression of ORF26 when added individually to BC-3 cells. Particularly, ORF26 expression stimulated with IFN-γ and TNF-a respectively, increased 6. 1 and 2. 5-fold(from real-time PCR results)at the 7th day when compared with untreated BC-3 cells.Meanwhile, about 20% of IFN-γ stimulated BC-3 cells expressed ORF 5 9 at the 7th day as compared with 1. 5 % of untreated BC-3 cells when IHC staining was employed. In addition, viral particles of HHV-8 were readily identified in BC-3 cells stimulated with IFN-γ at the 7th day with EM analysis.Conclusion: TNF-a and recombinant cytokines beingsimilar to those produced by HIV-1 infected T Cellscould really induce HHV-8 lytic cycle replication inBC-3 cells, another cell line of PEL.