IMMUNE CELL ACTIVATION BY A UNIQUE LENTIVIRUS NEF ALLELE

国际动物保护协会正在进行猴免疫缺陷病毒(SIV)的研究 赞助实验室，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 在艾滋病毒/艾滋病领域成功的独立研究人员。这个 这个项目的实验方面主要集中在SIVsmmPBj14上，它 在体内引起严重的急性疾病，并有效地触发 静息状态下猿猴淋巴细胞的体外增殖。这些属性 依赖于免疫受体酪氨酸激活的存在 SIVsmmPBj14 Nef中的基序(ITAM)。以下假设将是 检测：(I)小鼠的淋巴增殖诱导表型 SIVsmmPBj14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，而且在单核细胞上也是如此，这会增强病毒 在这些细胞中复制。具体目标1将检查是否良好- Nef的定义区域，包括保守的SH3结合基序(PxxP)， 是SIVsmmPBj14在猿猴体内有效复制所必需的 单核细胞，并用于诱导淋巴增殖。目标2将 评估基因nef阴性的反式互补 SIVsmmPBj14可恢复Nef的功能活性(由 诱导鸡淋巴细胞增殖和病毒复制效率的研究 原代单核细胞)。最后，目标3将检查是否表达 SIVsmmPBj14 Nef改变原发灶细胞内信号通路 单核细胞，以及这是否依赖于ITAM。重点将是 放在细胞蛋白激酶的激活和转录上 可能增强病毒基因表达的因素。总体而言， 拟议中的实验有望阐明致病机理 促进艾滋病发展的机制。

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.