Hantavirus: Hemorrhagic Fever Immunopathogenesis

汉坦病毒：出血热免疫发病机制

• 批准号: 7698541
• 负责人: Daniel H. Libraty
• 金额: $ 39.24万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-15 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7698541
• 关键词: Activation Analysis; Acute; Address; Aerosols; Affect; Antibodies; Antigen-Antibody Complex; Antigens; Area; Behavior; Biological Assay; Blood Cells; Blood Vessels; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; CD8B1 gene; Categories; Cell physiology; Cells; Class; Clinical; Clone Cells; Cohort Studies; Cytotoxic T-Lymphocytes; Dendritic Cells; Detection; Development; Disease; Endothelial Cells; Extravasation; Fever; Finland; Flow Cytometry; Functional disorder; Gene Expression; Genomics; Goals; Hantavirus; Hantavirus Infections; Hantavirus Pulmonary Syndrome; Hemorrhagic Fever with Renal Syndrome; Human; Immune; Immune response; Immunoassay; Immunologic Markers; Immunologics; In Vitro; Interferon Type I; Kidney; Kinetics; Lead; Leukocytes; Lung; Maps; Mediating; Mediator of activation protein; Myalgia; National Institute of Allergy and Infectious Disease; Organ; Pathway interactions; Pattern; Peptides; Peripheral Blood Mononuclear Cell; Puumala virus; RNA Viruses; Reverse Transcriptase Polymerase Chain Reaction; Sampling; Screening procedure; Severity of illness; Shapes; Specificity; Staging; Staining method; Stains; Syndrome; T-Lymphocyte; T-Lymphocyte Epitopes; Techniques; Therapeutic; Thrombocytopenia; Urine; Vaccines; Viral; Viral Antigens; Viral Hemorrhagic Fevers; Viral Load result; Viral Proteins; Virus; Virus Diseases; base; biodefense; cell behavior; cell growth regulation; chemokine; cytokine; in vivo; novel; pathogen; prospective; response; synthetic peptide

Hantaviruses are RNA viruses that cause hantavirus pulmonary syndrome (HPS) and hemorrhagic fever with renal syndrome (HFRS). HPS and HFRS are characterized by fever, myalgia, rapid onset of a vascular leak syndrome, hemoconcentration, and thrombocytopenia. In HPS, the lung is the prominent target organ; while, in HFRS, the kidney is the prominent target organ. Hantaviruses are NIAID category A priority pathogens with regards to biodefense, as they can produce severe, potentially fatal, diseases, are transmitted by aerosol, and do not have effective vaccines or specific therapeutics. The goal of this project is to understand the immunologic mechanisms that lead to HFRS. Several lines of evidence suggest that HFRS is not caused by direct cytopathic effects of hantaviruses, but rather by exuberant host immunopathological responses. This project will rely on samples provided from a prospective cohort study of Puumala (PUU) virus infections, a HFRS-associated hantavirus in Finland. The first aim will be to characterize dendritic cell functions and humoral immune responses that affect the PUU virus burden, using flow cytometry, antibody detection assays, and quantitative viral RT-PCR. The second aim will be to analyze the patterns and temporal regulation of cellular immune responses throughout acute PUU virus infection. ELISAs, multiplex immunoassays, quantitative RT-PCR, and genomic screening techniques will be used to examine immune response mediators in a comprehensive fashion, along with virus levels and disease severity. The third aim will be to characterize the antigen specificity and behavior of T lymphocyte responses during and after PUU virus infection. CD8+ and CD4+ T cell epitopes from PUU virus proteins will be identified using cell cloning techniques, ELISPOTs, cytotoxic T lymphocyte (CTL) assays, and mapping with overlapping synthetic peptides. Effector mechanisms of vascular leakage will be studied by examining interactions between endothelial cells and PUU virus-specific T cell clones. Peptide stimulation with intracellular cytokine staining and peptide-HLA Class I and II tetramers will be used to identify and quantify antigen-specific T cell responses across a spectrum of PUU virus disease. Elucidation of the immunopathogenetic mechanisms in PUU virus infection will contribute to the development of effective vaccine strateaies and immune-based therapies of HFRS and HPS.

汉坦病毒是引起汉坦病毒肺综合征(HPS)和 肾综合征出血热(HFRS)。HPS和HFRS的特征是发热、肌肉疼痛、迅速 出现血管渗漏综合征、血液浓缩和血小板减少症。在HPS中，肺是 突出靶器官；而在肾综合征出血热中，肾脏是突出靶器官。汉坦病毒是NIAID A类病原体在生物防御方面的优先地位，因为它们可以产生严重的、潜在的致命的、 疾病是通过气雾剂传播的，没有有效的疫苗或特定的治疗方法。 该项目的目标是了解导致肾综合征出血热的免疫学机制。几个 一系列证据表明，肾综合征出血热不是由汉坦病毒的直接细胞病变效应引起的，而是由 旺盛的宿主免疫病理反应。该项目将依赖于从潜在客户提供的样本 芬兰与肾综合征出血热相关的汉坦病毒--普玛拉(PUU)病毒感染的队列研究。第一个目标是 为了表征影响PUU病毒载量的树突状细胞功能和体液免疫反应， 使用流式细胞术、抗体检测和病毒定量RT-PCR。第二个目标将是 分析急性PUU病毒细胞免疫应答的模式和时间调节 感染。ELISA、多重免疫分析、定量RT-PCR和基因组筛选技术将 用于全面检查免疫反应介质，以及病毒水平和 疾病的严重性。第三个目标将是表征T淋巴细胞的抗原特异性和行为 PUU病毒感染期间和感染后的反应。PUU病毒蛋白的CD8+和CD4+T细胞表位将 通过细胞克隆技术、ELISPOTS、细胞毒性T淋巴细胞(CTL)检测和地图绘制进行鉴定 有重叠的合成多肽。将通过检查来研究血管渗漏的效应机制 内皮细胞与PUU病毒特异性T细胞克隆的相互作用。多肽刺激 将使用细胞内细胞因子染色和多肽-HLAI和II四聚体来鉴定和定量 PUU病毒谱中的抗原特异性T细胞反应。澄清： 免疫致病机制在PUU病毒感染中的作用 HFRS和HPS的疫苗策略和基于免疫的治疗。