IMMUNE CELL ACTIVATION BY A UNIQUE LENTIVIRUS NEF ALLELE

独特的慢病毒 NEF 等位基因激活免疫细胞

• 批准号: 6349756
• 负责人: LINDA E WHETTER
• 金额: $ 9.87万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-02-15 至 2002-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6349756
• 关键词: T lymphocyte; biological signal transduction; gene complementation; leukocyte activation /transformation; lymphocyte proliferation; molecular cloning; monocyte; protein kinase; regulatory gene; simian AIDSs; simian immunodeficiency virus; tissue /cell culture; transcription factor; virus genetics; virus replication

The PI is conducting research on simian immunodeficiency virus (SIV) in the sponsors laboratory, and sets forth a specific series of experiments and career development activities designed to result in her becoming a successful, independent researcher in the HIV/AIDS field. The experimental aspects of this project focuses on SIVsmmPBj14, which causes severe acute disease in vivo, and efficiently triggers the proliferation of resting simian lymphocytes in vitro. These properties depend on the presence of an immunoreceptor tyrosine-based activation motif (ITAM) in SIVsmmPBj14 Nef. The following hypotheses will be tested: (i) that the lymphoproliferation-inducing phenotype of SIVsmmPBj14 is dependent not only on the Nef ITAM, but also on additional determinants in Nef, and (ii) that the Nef ITAM has effects not only on T cells, but also on monocytes, which enhance viral replication in these cells. Specific aim 1 will examine whether well- defined regions of Nef, including a conserved SH3-binding motif (PxxP), are required for SIVsmmPBj14 to replicate efficiently in simian monocytes, and for the induction of lymphoproliferation. Aim 2 will assess whether trans-complementation of genetically nef-negative SIVsmmPBj14 can restore the functional activity of Nef (as measured by induction of lymphoproliferation and efficiency of viral replication in primary monocytes). Finally, aim 3 will examine whether expression of SIVsmmPBj14 Nef alters intracellular signaling pathways in primary monocytes, and whether this is dependent on the ITAM. Emphasis will be placed on activation of cellular protein kinases, and transcription factors which may enhance viral gene expression. Overall, the experiments proposed are expected to shed light on the pathogenic mechanisms which contribute to the development of AIDS.

PI正在进行猴免疫缺陷病毒（SIV）的研究， 发起人实验室，并提出了一系列具体的实验， 和职业发展活动，旨在使她成为一个 艾滋病领域的独立研究员。的 该项目的实验方面集中在SIVsmmPBj 14上， 在体内引起严重的急性疾病，并有效地触发 体外静止猿猴淋巴细胞的增殖。这些属性 依赖于基于免疫受体酪氨酸的激活的存在 SIVsmmPBj14 Nef.以下假设将 测试：（i）淋巴细胞增殖诱导表型 SIVsmmPBj 14不仅依赖于Nef ITAM，还依赖于 Nef中的其他决定因素，以及（ii）Nef ITAM具有影响 不仅在T细胞上，而且在单核细胞上， 在这些细胞中复制。具体目标1将审查是否良好- Nef的确定区域，包括保守的SH 3结合基序（PxxP）， SIVsmmPBj 14在猿中有效复制所必需的 单核细胞和用于诱导淋巴细胞增殖。目标2将 评估基因nef阴性的反式互补是否 SIVsmmPBj 14可以恢复Nef的功能活性（如通过免疫组织化学法测量的）。 诱导淋巴细胞增殖和病毒复制效率 初级单核细胞）。最后，目标3将检查是否表达 SIVsmmPBj 14 Nef改变原发性肝癌细胞内信号通路 单核细胞，以及这是否依赖于ITAM。重点将 置于细胞蛋白激酶的激活和转录 可能增强病毒基因表达的因子。总体看 提出的实验有望揭示致病性 这些机制有助于艾滋病的发展。