IMMUNOPATHOLOGICAL ANALYSIS OF DENGUE HEMORRHAGIC FEVER/DENGUE SHOCK SYNDROME

登革热病毒感染是#年发病率和死亡率的严重原因。 世界上许多地区。严重并发症的发病机制 登革病毒感染，登革出血热(DHF)对 阐明DHF的预防和治疗。流行病学研究 已经表明，DHF在第二次世界大战期间更常见 感染不同血清型的登革热病毒 最初的感染，并假设DHF是由 免疫病理机制。我们假设，增强的感染 登革热病毒抗体复合体诱导单核细胞显著活化 登革热病毒特异性的CD4和CD8T细胞及其高表达 导致DHF的细胞因子水平。 这个项目的目标是定义免疫病理机制， 使用分子免疫学技术诱导DHF。没有动物模型是 可用于研究DHF；因此，使用人体受试者的研究是 必填项。我们将通过分析来阐明免疫病理机制 弥漫性心衰患者外周血单个核细胞和血浆的检测 或在急性期及之后患有无并发症的登革热(D) 恢复。我们将：1)测定外周血中淋巴因子mRNA的水平 体内激活的CD8 T细胞，2)确定登革热病毒特异性 通过建立CD4T细胞，DHF患者的CD4=T细胞为Th1或Th2 克隆，3)确定登革热病毒特异性CD8=T细胞克隆是否也 产生一组具有特征的淋巴因子，并分析T细胞受体 Valpha和Vbeta基因的使用，并确定T细胞激活是否 体内寡克隆。 单核细胞是最容易复制登革热病毒的人类细胞。 我们将测定患者单核细胞中细胞因子mRNA的水平。 使用DHF或D。此外，我们将确定感染性水平 登革病毒抗体复合体在DHF患者血浆中的表达。这个 D和DHF患者病毒分离株复制能力的比较 在人类单核细胞中进行测试将在这个项目中进行测试。如果 检测到显著的增长差异，我们将利用这一点作为 选择典型菌株进行基因组测序的生物标记 项目3.我们将定义体内的免疫反应 在DHF中，基于对DHF或D患者样本的分析。 这些分析将提供有关抗体的基本免疫学数据。 以及对登革热病毒的T细胞反应，这是发展登革热所必需的 安全有效的登革热疫苗。

Dengue virus infections are a serious cause of morbidity and mortality in many areas of the world. The pathogenesis of severe complications of dengue virus infection, dengue hemorrhagic fever (DHF) is important to elucidate for prevention and treatment of DHF. Epidemiological studies have shown that DHF is much more commonly observed during secondary infections with a different serotype of dengue virus from that which caused the primary infection, and it is assumed that DHF is caused by immunopathological mechanisms. We hypothesize that enhanced infection of monocytes by dengue virus-antibody complexes results in marked activation of dengue virus-specific CD4+ and CD8+ T cells and the production of high levels of cytokines which lead to DHF. The goal of this project is to define immunopathological mechanisms which induce DHF, using molecular immunological techniques. No animal models are available to study DHF; therefore, research using human subjects is required. We will elucidate immunopathological mechanisms by analyzing peripheral blood mononuclear cells (PBMC) and plasma of patients with DHF or with uncomplicated dengue fever (D) during the acute phase and after recovery. We will; 1) determine the levels of lymphokine mRNA in CD4+ and CD8+ T cells activated in vivo, 2) determine whether dengue virus-specific CD4=T cells in patients with DHF are Th1 or Th2 by establishing CD4+T cell clones, 3) determine whether dengue virus-specific CD8=T cell clones also produce a characteristic set of lymphokines, and 4) analyze T cell receptor Valpha and Vbeta gene usage and determine whether T cell activation is oligoclonal in vivo. Monocytes are the most permissive human cells for dengue virus replication. We will determine the levels of cytokine mRNA in monocytes from patients with DHF or D. Furthermore, we will determine the levels of infectious dengue virus-antibody complexes in the plasma of patients with DHF. The comparative ability of virus isolates from D and DHF patients to replicate in human monocytes will be tested in tested in this project. If significant differences in growth are detected, we will use this as a biologic marker for selecting typical strains for genome sequencing in project 3. We will define the immunological responses in vivo which result in DHF, based on these analyses of samples from patients with DHF or D. These analyses will provide basic immunological data concerning antibody and T cell responses to dengue virus that are needed for the development of safe and effective dengue vaccines.