Gene Expression Induced by HIV-1 and Chemokine Receptor

除了趋化作用外，C-X-C和C-C趋化因子在T细胞活化和许多淋巴细胞生物学反应中起中介作用。有关下游信号通路的详细信息对于了解趋化因子在正常生理和炎症中的作用是必要的。我们已经建立了几个包含大约3000个已知人类基因的聚焦基因芯片，这些基因由包括淋巴样细胞在内的各种细胞类型表达和分泌，参与细胞生长、信号转导、效应功能、CD分子、新陈代谢和细胞凋亡。利用差异显示分析和商业基因芯片的初步研究表明，表达内源性趋化因子受体或转基因趋化因子受体的T细胞在配体处理后可以表达各种已知和未知的基因。已鉴定的基因包括硫氧还蛋白、CA150、Flotillin、铁蛋白重链和各种细胞因子。RT-PCR和Western印迹分析证实了这些差异表达。其他研究正在测试各种HIV-1病毒分离株、gp120蛋白、细胞因子和趋化因子直接诱导年轻人和老年人淋巴细胞和神经细胞基因表达的能力。我们认为，通过CD4和/或趋化因子受体分子激活的转录信号对于优化HIV-1的感染性和繁殖以及正常的淋巴细胞黏附和迁移是必需的。使用我们定制的人和鼠基因芯片和寡核苷酸微阵列基因芯片，我们已经检测了趋化因子受体连接后或病毒感染后超过22K的已知和未知基因的表达。我们相信，对诱导或抑制基因的鉴定和检测不仅可以深入了解HIV的发病机制，还可以阐明炎症的分子机制和在老年淋巴细胞中观察到的各种信号缺陷。我们目前正在确认和鉴定T细胞在SDF-1、MIP-3、RANTES、gp120和HIV-1病毒刺激下迁移后高表达的几个基因。更好地理解不同趋化因子受体连接诱导的转录信号可能提供一种手段来剖析这些趋化因子诱导细胞迁移和激活的途径，以及任何在HIV进入和复制中重要的宿主转录信号。

Besides chemotaxis, C-X-C and C-C chemokines function as mediators in T-cell activation and in many lymphocyte biological responses. Detailed information about downstream signaling pathways is necessary to understand the role of chemokines in normal physiology and inflammation. We have built several focused cDNA chips containing approximately 3000 known human genes that are expressed and secreted by a variety of cell types including lymphoid cells and participate in cell growth, signal transduction, effector functions, CD molecules, metabolism and apoptosis. Initial studies using differential display analysis and commercial cDNA chips have demonstrated that T cells expressing either endogenous chemokine receptors or transfected chemokine receptors are induced to express a variety of known and unknown genes post ligand treatment. A number of genes have been identified including thioredoxin, CA150, flotillin, ferritin heavy chain, and various cytokines. These mRNA expression difference have since been verified by RT-PCR and Western blot analysis. Additional studies are underway examining the ability of various HIV-1 viral isolates, gp120 proteins, cytokines, and chemokines to directly induce gene expression in young and old human lymphocytes and neuronal cells. We believe that active transcriptional signals through CD4 and/or chemokine receptor molecules are required for optimal HIV-1 infectivity and propagation as well as for normal lymphocyte adhesion and migration. Using our customized human and murine cDNA and oligonucleotide microarray gene chips, we have examined the expression of over 22K known and unknown genes induced post chemokine receptor ligation or viral infection. We believe that the identification and examination of induced or suppressed genes will not only provide insight into HIV pathogenesis but may also elucidate the molecular mechanisms of inflammation and the various signaling defects observed in aged lymphocytes. We are currently confirming and characterizing several genes highly expressed in T cells after migration in response to or stimulation with SDF-1, MIP-3, RANTES, gp120 and HIV-1 virus. A greater understanding of the transcriptional signals differentially induced by the ligation of various chemokine receptors may provide a means to dissect the pathways by which these chemoattractants induce cell migration and activation as well as any host transcriptional signals important in HIV entry and replication.