HIV infection and innate defense mechanisms in dendritic cells

异性传播是世界范围内HIV-1感染的主要方式。因此，了解粘膜早期的先天免疫反应对于设计预防感染的新策略是至关重要的。 令人信服的证据表明，在艾滋病毒-1感染后的最初几天和几周内发生的免疫学事件是影响艾滋病毒-1/艾滋病病程的关键决定因素。先天免疫反应未能限制HIV-1感染的机制目前正在紧张的研究中。该计划项目的目标是使用系统生物学方法剖析针对HLV-1的先天免疫反应中的早期事件。我们的计划(项目4)将使用一个原代DC-T细胞系统来测试这一假设，即HIV-1通过干扰树突状细胞(DC)的功能，特别是在这些细胞中诱导I型干扰素(IFN)来操纵人类先天免疫反应的动力学。 我们推测HIV-1延缓了DC的成熟，并且一个或多个HIV-1蛋白编码一种干扰素拮抗剂。在特定的目标1中，我们将确定HIV-1感染和干扰素/模式识别反应信号相互之间的相互影响。在特定目标#2中，我们将评估新发现的细胞限制因子在病毒感染的背景下在DC中的作用。我们将评估病毒复制和从树突状细胞转移到T淋巴细胞的效率，树突状细胞激活的模式和干扰素/PRR信号通路。我们将使用慢病毒转导系统下调或过度表达选定的宿主因子(50-100)，以测试它们在髓系DC谱系和T淋巴细胞中获得或丧失功能的影响。在特定的目标#3中，我们将使用一系列不同亚型的原始病毒分离株、删除辅助基因的HIV-1全长分子克隆来鉴定可能的DC成熟和干扰素产生的病毒抑制物。我们将通过将单个附件和调节性HIV-1基因插入重组新城疫病毒(NDV)载体来证实和扩大我们的发现，这些重组新城疫病毒载体可以诱导快速而强大的天然免疫反应。该项目结合了安娜·费尔南德斯博士的互补专业领域- 塞斯玛和维维亚娜·西蒙博士。Fernandez-Sesma博士在原代人类DC以及不同病毒(如流感、登革热(DENV)和新城疫病毒)在这些细胞中启动免疫反应方面拥有丰富的经验。Simon博士在HIV-1分子病毒学和影响HLV-1复制的宿主因素(如AP0BEC3)方面拥有丰富的专业知识。我们项目的结果将作为项目6中生成的数学模型的原始数据。本研究项目将确定项目1和2中确定的限制因素在DC和T淋巴细胞中的信号、DC成熟、先天性免疫反应和病毒抑制方面的作用，这两个主要细胞群与粘膜免疫最相关。

Heterosexual transmission is the dominant mode of HIV-1 acquisition woridwide. Understanding the eariy innate immune response in the mucosa is, thus, essential for devising novel strategies to prevent infection. Compelling evidence suggests that immunological events occurring during the first days and weeks after HIV-1 infection are critical determinants shaping the course of HIV-1/AIDS disease. The mechanisms that underiie the failure of innate immune responses to restrict HIV-1 infection are currently under intense investigation. The goal of this program project is to dissect the early events in the innate immune response directed at HlV-1 using a systems biology approach. Our proposal (project 4) will use a primary DC-T cell system to test the hypothesis that HIV-1 manipulates the kinetics of human innate immune responses by interfering with dendritic cell (DC) function, in particular, the induction of type I interferon (IFN) in those cells. We speculate that HIV-1 delays DC maturation and that one or more of the HIV-1 proteins encodes an IFN antagonist. In specific aim #1 we will determine the reciprocal impact of HIV-1 infection and IFN/pattern recognition response signaling on each other. In specific aim #2 we will evaluate the role of newly identified cellular restriction factors in DCs within the context of viral infection. We will assess the efficiency of viral replication and transfer from dendritic cells to T lymphocytes, the pattern of DC activation and the IFN/PRR signaling pathways. We will use lentiviral transduction systems to down-regulate or over-express selected host factors (50-100) to test the effect of their gain or loss of function in myeloid DC lineages and T lymphocytes. In specific aim #3 we will identify putative viral inhibitors of DC maturation and IFN production using a series of primary viral isolates of different subtypes, HIV-1 full-length molecular clones deleted of accessory genes. We will confirm and expand our findings by inserting single accessory and regulatory HIV-1 genes into recombinant Newcastle Disease Virus (NDV) vectors which induce rapid and strong innate immune responses. This project combines the complementary areas of expertise of Dr. Ana Fernandez- Sesma and Dr. Viviana Simon. Dr. Fernandez-Sesma has extensive experience with primary human DCs and the initiation of immune responses in those cells by different viruses, such as Influenza, Dengue (DENV) and NDV. Dr. Simon has great expertise in HIV-1 molecular virology and host factors influencing HlV-1 replication, such as AP0BEC3. The results of our project will serve as raw data forthe mathematical models generated in project 6. This research project will determine the role of the restriction factors identified in project 1 and 2 on signaling, DC maturation, innate immune responses and viral inhibition in DCs and Tlymphocytes, both primary cell populations most relevant to mucosal immunity.