CELLULAR FACTORS THAT CONTROL VIRAL EXPRESSION AND REGULATION

我们正在研究介导信号的细胞因子的作用， 在病毒基因表达的调节中的转导。 我们分析 细胞癌基因c-fos、c-myc和v-src（a 酪氨酸激酶）在人外周血中的表达 淋巴细胞（PBL）和H9和U937细胞。 静止的PBLs用HIV-1感染，使用无血清培养基， 存在或不存在IL-2。 在感染后的不同时间， 分析RNA中病毒和癌基因序列的存在， 将细胞固定以通过流式细胞术分析抗原。 20折 c-fos RNA增加和c-fos抗原增加4-5倍， 在感染后2至22小时之间观察。 这一增长 放线菌素D抑制。 在这些相同的细胞中，HIV-1序列被 在2小时时首次在基因组DNA中检测到病毒转录物， 感染后4小时。 我们的初步结果表明， 这些细胞癌基因中的一些的表达的增强可 在调节受感染细胞中的病毒复制中起作用。 类似 对感染HIV-2的HUT-78和CEM细胞的研究正在进行中。 评价转染的c-fos和c-jun的作用的实验（另一个 细胞癌基因）在艾滋病毒基因组反式激活中的作用 中求进工作总 凝胶阻滞试验和紫外交联研究正在进行中。 研究这些因素。 我们还计划研究 抗HIV药物对HIV-1和HIV-2感染细胞中细胞激酶的作用。 这些研究将使我们能够进一步确定病毒的阶段 这些基因产物可能发挥作用的复制。

We are investigating the role of cellular factors that mediate signal transduction in the regulation of viral gene expression. We analyzed the expression of the cellular oncogenes c-fos, c-myc and v-src ( a tyrosine kinase expressed in T-cells) in human peripheral blood lymphocytes (PBLs) and H9 and U937 cells following infection with HIV-1. Quiescent PBLs were infected with HIV-1 using serum-free medium in the presence or absence of IL-2. At various times after infection DNA and RNA were analyzed for the presence of viral and oncogene sequences and cells were fixed for analysis of antigen by flow cytometry. A 20 fold increase in c-fos RNA and a 4-5 fold increase in c-fos antigen was observed between 2 and 22 hours post-infection. This increase was inhibited by actinomycin D. In these same cells, HIV-1 sequences were first detected in genomic DNA at 2 hours and viral transcripts between 2 and 4 hours post-infection. Our preliminary results suggest that enhancement of the expression of some of these cellular oncogenes may play a role in regulating viral replication in infected cells. Similar studies are in progress with HUT-78 and CEM cells infected with HIV-2. Experiments evaluating the role of tranfected c-fos and c-jun (another cellular oncogene) in the transactivation of the HIV genome are in progress. Gel retardation assays and UV cross linking studies are being performed to study these factors. We also plan to study the effects of anti-HIV agents on cellular kinases in HIV-1 and HIV-2 infected cells. These studies will enable us to further define the stages of viral replication at which these gene products may function.