INVESTIGATION OF AUTOSTIMULATORY PATHWAYS REGULATING HIV

描述：巨噬细胞在HIV-1病毒的形成中起关键作用 感染和发展为艾滋病。被感染的巨噬细胞是长寿的 病毒在许多组织中的蓄水池。此外，这些细胞产生 可能影响血管功能的炎性和细胞毒性因素 邻近的细胞和组织。这项提案的总体目标是 调查导致入侵的细胞和分子事件 内皮细胞和巨噬细胞在这一过程中的作用。调查员 提出转录激活物C/EBPbeta参与了 HIV-1感染过程中巨噬细胞基因表达失调性改变 巨噬细胞-内皮细胞的相互作用，并有助于 有利于HIV-1复制的环境。调查员使用 原代巨噬细胞与人脐静脉内皮细胞共培养 (HUVEC)以更好地了解这些互动如何影响 HIV-1复制，以及鉴定可溶性因子和细胞表面 调节这些反应的分子。调查员还将确定 巨噬细胞能否诱导内皮细胞凋亡及相关基因的表达 内皮细胞表面的磷脂酰丝氨酸(PS)。实验将是 以确定PS是否有助于招募受感染的人 巨噬细胞进入血管内皮细胞，持续破坏血管内皮细胞。 最后，将通过表征来直接测试C/EBPbeta的重要性 潜伏感染和未感染的单核细胞系 过度表达C/EBPbeta或功能上与之相互作用的显性负面嘴唇 内皮细胞。

DESCRIPTION: Macrophages play a critical role in the establishment of HIV-1 infection and progression to AIDS. Infected macrophages serve as long-lived reservoirs for virus in many tissues. Furthermore, these cells produce inflammatory and cytotoxic factors that may influence the function of neighboring cells and tissues. The overall goal of this proposal is to investigate the cellular and molecular events that lead to breaching of the endothelilium and the role of macrophages in this process. The investigator proposes that the transcriptional activator C/EBPbeta is involved in disregulated macrophage gene expression during HIV-1 infection, altering macrophage-endothelial cell interactions, and contributing to a micro environment that is favorable for HIV-1 replication. The investigator uses primary macrophages co-cultured with human umbilical vein endothelial cells (HUVEC) to gain a better understanding of how these interactions influence HIV-1 replication, as well as to identify soluble factors and cell surface molecules that mediate these responses. The investigator will also determine whether macrophages can induce endothelial cell apoptosis and expression of phosphatidylserine (PS) on the endothelial cell surface. Experiments will be conducted to determine whether PS contributes to the recruitment of infected macrophages to the endothelium and perpetuates endothelial cell destruction. Finally, the importance of C/EBPbeta will be directly tested by characterizing the ability of latently infected and uninfected monocytic lines that overexpress C/EBPbeta or a dominant negative LIP to functionally interact with endothelial cells.