Mechanism of cell-associated HIV-1 transmission

 描述(申请人提供)：巨噬细胞是哨兵细胞，在广泛的先天和获得性免疫反应中发挥关键作用。巨噬细胞可以被HIV-1感染，因为巨噬细胞是长寿细胞，对病毒感染的细胞病变具有抵抗力，在整个疾病过程中对病毒库做出了重大贡献。巨噬细胞广泛存在于所有组织中，可通过细胞间连接将病毒高效地传播给CD4+T细胞。在这项提案中，我们将检验HIV-1在巨噬细胞中建立生产性感染的假设，以及从头开始病毒蛋白表达触发I型干扰素依赖的促炎反应。使用前病毒突变和表达策略，我们将确定诱导先天反应所必需的病毒决定因素。我们将利用免疫亲和纯化和质谱学方法来鉴定检测这些病原体决定簇的宿主因子(S)，以及该尚未确定的宿主因子与病毒决定簇结合时触发的信号级联反应。最后，我们将确定先天反应的诱导是否会增强干扰素刺激的基因，如CD169的表达，这些基因被HIV-1颠覆，以促进病毒通过感染突触传播到CD4+T细胞，这是全身病毒传播的一种巨噬细胞依赖机制。

 DESCRIPTION (provided by applicant): Macrophages are sentinel cells that play a pivotal role in a broad range of innate and adaptive immune responses. Macrophages can be infected by HIV-1 and because these are long-lived cells and are resistant to the cytopathic effects of virus infection, contribute significantly to the virus reservoir throughout the course of the disease. Macrophages are present ubiquitously in all tissues and can disseminate virus to CD4+ T cells across cell-to-cell junctions with high efficiency. In this proposal, we will test the hypothesis tat establishment of productive infection in macrophages by HIV-1 and de novo viral protein expression triggers type I IFN- dependent pro-inflammatory responses. Using a proviral mutagenesis and expression strategy, we will identify the viral determinants that are necessary for inducing innate responses. We will utilize immuno-affinity purification and mass spectrometry approaches to identify the host factor(s) that detects these pathogen determinants and the signaling cascades that are triggered upon engagement of the viral determinants by this yet to be identified host factor. Finally, we will determine if induction of innate responses enhances expression of interferon-stimulated genes, such as CD169, that are subverted by HIV-1 to facilitate virus spread to CD4+ T cells across infectious synapses, a macrophage-dependent mechanism of systemic virus dissemination.