MOLECULAR DETERMINANTS OF HIV EXPRESSION BY OPIATES

阿片类药物表达 HIV 的分子决定因素

• 批准号: 3213117
• 负责人: OM PRAKASH
• 金额: $ 17.53万
• 依托单位: OCHSNER CLINIC FOUNDATION
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-08-01 至 1992-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3213117
• 关键词: HIV infections; T lymphocyte; acyltransferase; antisense nucleic acid; autoradiography; brain; clone cells; drug abuse; fusion gene; gene deletion mutation; gene expression; genetic manipulation; genetic regulatory element; genetic transcription; genetically modified animals; genome; hematopoietic tissue; human immunodeficiency virus; human tissue; laboratory mouse; morphine; naloxone; neurons; nucleic acid repetitive sequence; nucleoproteins; opiate alkaloid; protooncogene; tissue /cell culture; transfection

The retrovirus HIV (human immunodeficiency virus) has been identified as the causative agent in acquired immunodeficiency syndrome (AIDS). Infection is followed in many cases by a clinically quiescent or latent phase that appears to continue as long as host antiviral defense is intact. These findings raise the possibility htat host susceptibility factors (i.e., "environmental cofactors") may influence the progression of the disease. The identification of such cofactors is, obviously, of immense significance in the understanding and characterization of the pathogenesis of AIDS. An estimated 25% of all AIDS cases in the United States are drug abusers. Recently, the findings that various "drugs of abuse" (i.e., opiates) are immunomodulatory and immunocompromising has ld to the suggestion that opiates may serve as putative environmntal cofactors and influence the susceptibility and progression of HIV infection. Our goal is to define the relationship between opiates and the human immunodeficiency virus genome on a functional level. We will use molecular genetic approaches and both in vitro and in vivomodels to characterize the effects of opiates on the transcriptional regulation of HIV. To examine this question directly, we will use HIV-long terminal repeat-chloramphenicol acetyltransferase fusion genes (HIV/LTR-CAT). The HIV/LTR-CAT chimeric constructs will be transfected into both human clonal T-cells and human neural cells in order to establish appropriate in vitro models in which to examine this putative relationship. W have demonstrated that obth cell types are responsive to morphine in a naloxone-reversible manner at the level of gene expression. We will use deletion mutagenesis to define the opiate-responsive region of the HIV LTR. Since previous results from our laboratory have demonstrated a rapid and transient inductive effect of morphine on c-fos expression, we will also determine if Fos can function as an adaptor gene and link the short-term post-receptor effects of opiates to the activation HIV expression. Our hypothesis is based on the recent identification of two AP-1 transcriptional regulatory sequences within the HIV LTR and that the AP-1 sequence is required for the transcriptional activation of eukaryotic genes by a Fos-containing nucleoprotein complex. Finally, we have proposed to introduce HIV/LTR-CAT fusion genes into the germline of mice (i.e., transgenic mice) in order to establish appropriate in vivo models. We predict our findings will have important implications for the pathogenesis of AIDS in patients classified as drug abusers.

逆转录病毒HIV（人类免疫缺陷病毒）已被确定为 获得性免疫缺陷综合症（艾滋病）的病原体。 感染后，在许多情况下，临床上静止或潜伏 这一阶段似乎只要宿主的抗病毒防御持续， 完整 这些发现提高了htat宿主易感性的可能性 因素（即，“环境辅助因子”）可能会影响进展 的疾病。 这种辅因子的鉴定显然是 在理解和描述 艾滋病的发病机制。 据估计，美国25%的艾滋病病例 国家是吸毒者。 最近，研究发现，各种“药物的 滥用”（即，阿片类药物）具有免疫调节和免疫损害作用， 对于阿片类药物可能作为假定的麻醉剂的建议， 辅助因子和影响艾滋病毒的易感性和进展 感染 我们的目标是确定阿片类药物和 人类免疫缺陷病毒基因组的功能水平。 我们将 使用分子遗传学方法，在体外和体内模型中， 描述阿片类药物对转录调控的影响， 艾滋病。 为了直接研究这个问题，我们将使用艾滋病毒长终端 重复氯霉素乙酰转移酶融合基因（HIV/LTR-CAT）。 将HIV/LTR-CAT嵌合构建体转染到两个人中， 克隆T细胞和人神经细胞，以建立适当的 在体外模型中检查这种假定的关系。 W具有 表明obth细胞类型对吗啡有反应， 纳洛酮可逆的方式在基因表达水平。 我们将使用 缺失诱变以确定HIV的阿片类药物反应区 LTR 由于我们实验室以前的结果表明， 吗啡对c-fos表达的快速和短暂的诱导作用， 还将确定Fos是否可以作为一个衔接基因发挥作用， 阿片类药物对HIV激活的短期受体后效应 表情 我们的假设是基于最近发现的两种 HIV LTR内的AP-1转录调控序列， AP-1序列是转录激活 真核生物基因通过含Fos的核蛋白复合物。 最后我们 已经提出将HIV/LTR-CAT融合基因引入到 小鼠（即，转基因小鼠）以建立适当的体内 模型 我们预测，我们的发现将对以下方面产生重要影响： 将艾滋病患者的发病机制归类为吸毒者。