HIV infection and innate defense mechanisms in dendritic cells

HIV感染和树突状细胞的先天防御机制

基本信息

批准号：
8013195
负责人：
Ana Fernandez-Sesma
金额：
$ 47.69万
依托单位：
SALK INSTITUTE FOR BIOLOGICAL STUDIES
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-08-15 至 2015-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8013195
关键词：
Acute Address Affect Antiviral Agents Area Biological Biological Assay CD4 Positive T Lymphocytes Cell Line Cell Lineage Cell Maturation Cell physiology Cells Code Collaborations Data Defense Mechanisms Dendritic Cells Dendritic cell activation Dengue Disease Down-Regulation Enzyme-Linked Immunosorbent Assay Event Failure Genes Genetic Genome Goals HIV Infections HIV-1 HIV-2 vaccine Heterosexuals Human Immune response Immunity Infection Infection prevention Influenza Integration Host Factors Interferon Receptor Interferon Type I Interferons Invaded Investigation Kinetics Knowledge Length Measures Mediating Microarray Analysis Modeling Molecular Cloning Molecular Virology Mucosal Immunity Mucous Membrane Mutation Myelogenous Myeloid Cells Natural Immunity Newcastle disease virus Nucleic Acids Participant Pathway interactions Patients Pattern Pattern Recognition Physiologic pulse Physiological Population Production Protein Biosynthesis Proteins RNA Viruses Receptor Signaling Recombinants Repression Research Project Grants Role Series Shapes Signal Pathway Signal Transduction System Systems Biology T-Lymphocyte Testing Toll-like receptors Vaccines Validation Viral Virus Virus Diseases Virus Inhibitors adaptive immunity base chemokine clinical material cohort cytokine experience gain of function genome wide association study genome-wide immunogenicity inhibitor/antagonist loss of function mathematical model migration mutant novel novel strategies overexpression pathogen programs receptor research study response transmission process vector vif Gene Products

项目摘要

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.

异性传播是全球范围内 HIV-1 感染的主要方式。因此，了解粘膜的早期先天免疫反应对于设计预防感染的新策略至关重要。令人信服的证据表明，HIV-1 感染后最初几天和几周内发生的免疫事件是影响 HIV-1/AIDS 疾病进程的关键决定因素。目前正在深入研究导致先天免疫反应未能限制 HIV-1 感染的机制。该项目的目标是利用系统生物学方法剖析针对 HIV-1 的先天免疫反应的早期事件。我们的提案（项目 4）将使用初级 DC-T 细胞系统来检验 HIV-1 通过干扰树突状细胞 (DC) 功能（特别是 I 型干扰素的诱导）来操纵人类先天免疫反应动力学的假设(IFN) 在这些细胞中。我们推测 HIV-1 延迟 DC 成熟，并且一种或多种 HIV-1 蛋白编码 IFN 拮抗剂。在具体目标#1中，我们将确定 HIV-1 感染和 IFN/模式识别响应信号传导之间的相互影响。在具体目标#2中，我们将评估新发现的细胞限制因子在病毒感染背景下在树突状细胞中的作用。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、DC 激活模式和 IFN/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)，以测试它们在骨髓 DC 谱系和 T 淋巴细胞中功能获得或丧失的影响。在具体目标#3中，我们将使用一系列不同亚型的初级病毒分离株、删除辅助基因的HIV-1全长分子克隆来鉴定假定的DC成熟和IFN产生的病毒抑制剂。我们将通过将单个辅助和调节性 HIV-1 基因插入重组新城疫病毒 (NDV) 载体来证实和扩展我们的发现，该载体可诱导快速而强烈的先天免疫反应。该项目结合了 Ana Fernandez 博士的互补专业领域- Sesma 和 Viviana Simon 博士。 Fernandez-Sesma 博士在人类原代 DC 以及不同病毒（例如流感、登革热 (DENV) 和 NDV）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 HIV-1 复制的宿主因素（例如 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。该研究项目将确定项目 1 和 2 中确定的限制因素对 DC 信号传导、DC 成熟、先天免疫反应和病毒抑制的作用，以及T淋巴细胞是与粘膜免疫最相关的两种原代细胞群。