Human Beta-Defensin-1 in HSV-1 Innate Immunity

HSV-1 先天免疫中的人类 Beta-Defensin-1

• 批准号: 7632394
• 负责人: Lisa Kathleen Ryan
• 金额: $ 17.88万
• 依托单位: UNIV OF MED/DENT OF NJ-NJ MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-20 至 2009-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7632394
• 关键词: Acquired Immunodeficiency Syndrome; Address; Animal Model; Animals; Antiviral Agents; Area; Binding Sites; Biological Assay; Biological Process; Blood Circulation; CCR6 gene; CD4 Positive T Lymphocytes; Cells; Chemotaxis; Chloroquine; Clinical; Clinical Research; Conditioned Culture Media; Cytotoxic T-Lymphocytes; Data; Defensins; Dendritic Cells; Development; Disease; Endosomes; Environment; Epithelial; Exhibits; Future; Gene Expression; Genes; Genetic Transcription; Goals; Grant; HIV; Herpesvirus 1; Host Defense; Host Defense Mechanism; Hour; Human; Human Virus; Immune; Immune response; Immune system; Immunity; Immunocompromised Host; Immunologic Deficiency Syndromes; Immunology; In Vitro; Infection; Influenza; Interferons; Kinetics; Knowledge; Lead; Life; Link; Mediator of activation protein; Messenger RNA; Methods; Modeling; Molecular; Mus; Myelogenous; Natural Immunity; Natural Killer Cells; Nature; Opportunistic Infections; Oral; Organ Transplantation; Pathway interactions; Patients; Peptide Antibiotics; Peptides; Play; Predisposition; Prevention; Process; Production; Promoter Regions; Property; Range; Receptor Signaling; Recruitment Activity; Regulation; Research; Research Personnel; Role; Signal Induction; Simplexvirus; Staging; Stomatitis; Study models; TLR9 gene; Testing; Therapeutic; Toll-Like Receptor 2; Toll-like receptors; Transcriptional Activation; Tumor Necrosis Factor-alpha; Tumor Necrosis Factors; Up-Regulation; Viral; Virus; Virus Diseases; Work; antimicrobial drug; antimicrobial peptide; base; cell type; chemokine; chemotherapy; cytokine; design; human DEFB1 protein; human TNF protein; immune function; improved; in vitro Assay; in vivo; inhibitor/antagonist; lymph nodes; member; monocyte; novel; oral biology; pathogen; peripheral blood; receptor; research study; response; success; transcription factor

DESCRIPTION (provided by applicant): Viral infections caused by Herpes Simplex virus-1 (HSV-1), often occur as a result of a compromised immune system. The innate immune system is the first line of defense against prevention of these pathogens and consists of numerous soluble factors and cell types, including dendritic cells (DC) and natural killer cells. One type of DC, the plasmacytoid DC (PDC) is important in innate immune function through the production of interferon-? (IFN) induced by enveloped viruses like HSV-1. PDC have recently been shown to play a role in induction of Th1 immunity against influenza and HSV-1 and are recruited in mouse infection models to lymph nodes to stimulate antiviral cytotoxic T-cells. In AIDS patients, the lack of PDC in the circulation, along with depletion of CD4 T-lymphocytes is associated with the development of opportunistic infections, including viral infections. In addition to IFN, PDC produce chemokines and tumor necrosis factor-a upon HSV stimulation. We demonstrated that PDC also produce human beta-defensin-1 (hBD-1), an antimicrobial peptide that has antiviral activity and chemotactic properties towards cells transfected with CCR6 receptor. We hypothesize that an initial step in anti-viral immunity against HSV-1 is the production of hBD-1 in immune cells such as PDC. Our preliminary data show that HSV-1 induces increased hBD-1 mRNA and peptide in PDC within 2 hours of HSV-1 stimulation. Since PDC have potential importance in viral infections, it is essential to first understand the nature and mechanism of the viral induction of hBD-1 in PDC before determining its presence in clinical disease in patients. We hypothesize that HSV-1 directly induces relevant amounts of hBD-1 in PDC via stimulation of Toll-like receptor 9 and activation of transcription factors subsequently increasing hBD-1 gene transcription. This study proposes to explore the initial response pathway of HSV-1 induced hBD-1 mRNA and peptide in PDC, to develop a method to quantify hBD-1 induction in this rare DC subset, and to determine the transcriptional activation pathway of hBD-1 induction by HSV-1. In the first aim we will determine whether hBD-1 is stimulated by HSV-1 via a direct pathway or whether cytokines and chemokines influence the induction via an indirect mechanism. We will also study HSV-1 and PDC interactions using co- localization studies of virus and hBD-1 and whether hBD-1 requires endosomal acidification upon viral entry using chloroquine treatment of PBMC. To begin to address the biological function of hBD-1, chemotaxis studies will be done. We will examine TLR 2, 7 and 9 signaling, induction of interferon regulatory factors and identification of binding sites of transcription factors in the hBD-1 promoter region. The data from this grant will yield new information on a new mechanism for ¿-defensin regulation, different from hBD-2, and enhance the knowledge of the biological function of PDC and also enable future studies in vivo, designing animal studies that model HSV-1 infection or clinical studies. Herpes Simplex Virus-1 (HSV-1) is the cause of severe and often persistent opportunistic infections in immunocompromised patients, such as those undergoing chemotherapy, organ transplantation, and those with immunodeficiencies and HIV. The first line of defense against this and other viral pathogens is the innate immune system. One component of innate immunity is the production of antibiotic peptides such as ¿- defensins. The long-range goal of our research is to understand the role of ¿-defensins in the innate immune defense against viruses so that novel therapies can be developed from our findings.

描述（由申请人提供）： 由单纯疱疹病毒 1 (HSV-1) 引起的病毒感染通常是由于免疫系统受损而发生。先天免疫系统是预防这些病原体的第一道防线，由许多可溶性因子和细胞类型组成，包括树突状细胞 (DC) 和自然杀伤细胞。浆细胞样 DC (PDC) 是 DC 的一种类型，通过产生干扰素-α，在先天免疫功能中发挥重要作用。 (IFN) 由 HSV-1 等包膜病毒诱导。最近已证明 PDC 在诱导针对流感和 HSV-1 的 Th1 免疫中发挥作用，并在小鼠感染模型中被募集至淋巴结以刺激抗病毒细胞毒性 T 细胞。在艾滋病患者中，循环中 PDC 的缺乏以及 CD4 T 淋巴细胞的消耗与机会性感染（包括病毒感染）的发生有关。除了 IFN 之外，PDC 在 HSV 刺激下还产生趋化因子和肿瘤坏死因子-a。我们证明 PDC 还产生人 β-防御素-1 (hBD-1)，这是一种抗菌肽，对转染 CCR6 受体的细胞具有抗病毒活性和趋化特性。我们假设 HSV-1 抗病毒免疫的第一步是在 PDC 等免疫细胞中产生 hBD-1。我们的初步数据表明，HSV-1 刺激后 2 小时内，HSV-1 会诱导 PDC 中 hBD-1 mRNA 和肽的增加。由于 PDC 在病毒感染中具有潜在的重要性，因此在确定其在患者临床疾病中的存在之前，首先了解 PDC 中 hBD-1 病毒诱导的性质和机制至关重要。我们假设 HSV-1 通过刺激 Toll 样受体 9 和激活转录因子，随后增加 hBD-1 基因转录，直接诱导 PDC 中相应量的 hBD-1。本研究旨在探索PDC中HSV-1诱导hBD-1 mRNA和肽的初始反应途径，开发一种量化这种罕见DC亚群中hBD-1诱导的方法，并确定HSV-1诱导hBD-1的转录激活途径。第一个目标是确定 hBD-1 是否通过直接途径被 HSV-1 刺激，或者细胞因子和趋化因子是否通过间接机制影响诱导。我们还将通过病毒和 hBD-1 的共定位研究来研究 HSV-1 和 PDC 的相互作用，以及使用氯喹处理 PBMC 时，hBD-1 是否需要在病毒进入后进行内体酸化。为了开始解决 hBD-1 的生物学功能，将进行趋化性研究。我们将检查 TLR 2、7 和 9 信号传导、干扰素调节因子的诱导以及 hBD-1 启动子区域中转录因子结合位点的鉴定。这笔资助的数据将产生关于不同于 hBD-2 的 β-防御素调节新机制的新信息，并增强对 PDC 生物学功能的了解，也使未来的体内研究、设计 HSV-1 感染模型的动物研究或临床研究成为可能。单纯疱疹病毒-1 (HSV-1) 是免疫功能低下患者（例如接受化疗、器官移植的患者以及患有免疫缺陷和艾滋病毒的患者）严重且经常持续的机会性感染的原因。针对这种病毒和其他病毒病原体的第一道防线是先天免疫系统。先天免疫的组成部分之一是抗生素肽的产生，例如β-防御素。我们研究的长期目标是了解β-防御素在针对病毒的先天免疫防御中的作用，以便根据我们的发现开发新的疗法。