Dengue Infected Endothelial Cells Enhance Immune Cell Activation

登革热感染的内皮细胞增强免疫细胞激活

• 批准号: 8385024
• 负责人: Erich R Mackow
• 金额: $ 19.63万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-01 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8385024
• 关键词: Address; Anaphylatoxins; Antibodies; Antibody Formation; Arteries; Autopsy; B-Lymphocytes; Blood; Blood Platelets; Blood Vessels; Blood capillaries; CXCL10 gene; CXCL11 gene; Capillary Permeability; Cell Degranulation; Cell Line; Cell Maturation; Cell Proliferation; Cell Surface Receptors; Cells; Chemotaxis; Complement; Complement 3a; Complement 5a; Complement Activation; Culicidae; Dendritic Cells; Dengue; Dengue Hemorrhagic Fever; Dengue Shock Syndrome; Dengue Virus; Disease; Disease model; Edema; Endothelial Cells; Endothelium; Extravasation; Figs - dietary; Hemorrhage; Human; Immune; Immune Cell Activation; Immune Targeting; Immune response; Immunoperoxidase Technics; In Vitro; Infection; Inflammation; Interleukin-7; Left; Leukocytes; Liquid substance; Liver; Lung; Mediating; Monitor; Mus; Natural Killer Cells; Pathogenesis; Pathway interactions; Patients; Peripheral; Peripheral Blood Lymphocyte; Permeability; Plasma; Play; Process; Production; Properdin; Proteins; RANTES; Recruitment Activity; Role; Sampling; Serotyping; Shock; Spleen; Staining method; Stains; Surface; T cell activating factor; T-Lymphocyte; Vascular Permeabilities; Viral; Viral Antigens; Viral Pathogenesis; Viremia; Virus; Virus Activation; Virus Diseases; capillary; chemokine; complement system; cytokine; in vivo; irofulven; mast cell; mortality; mouse model; neutrophil; release factor; response; virus pathogenesis

DESCRIPTION (provided by applicant): Dengue fever (DF), dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS) are diseases caused by 4 serotypes of dengue virus (DV). Edema and hemorrhagic disease found in DHF/DSS result from vascular leakage, and these severe manifestations are enhanced by preexisting DV antibodies and subsequent infection by a second DV serotype. Thus an immune enhanced disease process contributes to increased endothelial cell (EC) permeability in DHF/DSS cases. DVs infect peripheral human leukocytes, dendritic cells and endothelial cells, and mechanisms by which DVs enhance vascular permeability are just beginning to be define. The ability of DVs to infect immune cells has resulted in many studies on infection of peripheral blood lymphocytes (PBLs) and their chemokine responses. Both are clearly important to viral pathogenesis and are readily studied in blood from humans or DV infected murine disease models. Although ECs are more difficult to study in vivo, they form the primary fluid barrier of the vasculature, and ultimately edema or hemorrhagic disease result from altering barrier functions of the endothelium. Postmortem studies of DV-infected patients and the AG129 mouse model demonstrate that ECs in the liver, lung, and spleen are infected. The ability of DVs to infect ECs provides a means for infection to alter capillary permeability, replicate and induce EC chemokine responses that activate and recruit immune cells to the endothelium. However, the contribution of DV infected ECs to immune enhancement and vascular permeability has yet to be factored into the DV disease process. DV infects ECs in culture, however prior in vitro studies were performed on ECs that were only 2-10% infected. In contrast, our recent studies of DV infected ECs were performed by synchronously infecting >80% of primary human ECs and monitoring viral and cellular responses. We found that DV productively infects primary human ECs, with a rapid increase in viral titer (~105/ml) 1 day p.i. Our findings suggest that DV infected ECs contribute to viremia, viral dissemination and the presentation of DV antigens on ECs that make them targets of antibodies and immune cells. Our analysis of EC responses to DV infection revealed the high level induction of chemokines that direct immune cell recruitment and activation. Cytokines CXCL10, CXCL11, IL-7, RANTES and BAFF are induced 337, 45, 128, 84 and 119 fold, respectively, in synchronously infected ECs by 24 hrs p.i. CXCL10/11 and RANTES secretion recruit and activate leukocytes and T-cells while IL-7 stimulates B, T and NK cell proliferation and T cell maturation. BAFF is a B and T cell activating factor that is released or expressed on the surface of activated ECs. DV infection also induced properdin (Factor P, 34-fold), which activates the alternative pathway (AP) complement system producing C3a/C5a: chemotactic anaphylatoxins which trigger localized inflammation, mast cell degranulation and vascular permeability. Importantly, high levels of C3a are present in severely ill dengue patients and associated with severe disease, plasma leakage and shock. These newly discovered EC responses indicate that ECs are not only targets of DV infection, but release factors that potentiate immune cell chemotaxis and permeability of the infected endothelium. Collectively these findings suggest that DV infected ECs contribute to viremia, enhanced immune responses and vascular permeability which are fundamental components of DHF and DSS. We propose to define EC responses to DV infection that enhance immune responses by activating and recruiting immune cells to the endothelium. PUBLIC HEALTH RELEVANCE: Dengue virus is carried by mosquitos and infects 50-100 million people/ year, causing a vascular leak disorder responsible for edema and hemorrhagic disease in 500,000-1,000,000/ year with a 5-30% mortality rate. Dengue results in an immune enhanced disease process whereby the 2nd infection is more severe from a distinct serotype. DV infects immune as well as endothelial cells that line capillaries and regulate edema and bleeding. We have found that DV infected endothelial cells elicit responses that enhance immune mediated responses that contribute to vascular leakage and contribute to increased virus production and spread. We address this process in endothelial cells in this study.

描述（由申请人提供）：登革热（DF），登革热大量热（DHF）和登革热综合征（DSS）是由4种登革热病毒（DV）引起的疾病。在DHF/DSS中发现的水肿和出血性疾病是由血管泄漏引起的，这些严重的表现通过先前存在的DV抗体和随后的第二DV血清型增强。因此，免疫增强的疾病过程有助于在DHF/DSS病例中增加内皮细胞（EC）渗透率。 DVS感染外周血细胞，树突状细胞和内皮细胞，以及DVS增强血管通透性的机制。 DV感染免疫细胞的能力导致了许多有关外周血淋巴细胞（PBL）及其趋化因子反应的感染的研究。两者对于病毒发病机理显然很重要，并且在人类或DV感染的鼠类疾病模型中很容易研究。尽管EC在体内更难研究，但它们构成了脉管系统的主要流体屏障，最终是由于内皮的屏障功能改变而导致的水肿或出血性疾病。 DV感染患者和AG129小鼠模型的死后研究表明，肝脏，肺和脾脏中的EC被感染。 DV感染ECS的能力为感染提供了一种改变毛细血管渗透性，复制并诱导EC趋化因子反应，从而激活和募集免疫细胞对内皮细胞。但是，DV感染EC对免疫增强和血管通透性的贡献尚未纳入DV疾病过程。 DV在培养中感染EC，但是对仅感染2-10％的EC进行了先前的体外研究。相反，我们最近对DV感染EC的研究是通过同步感染> 80％的原代人EC并监测病毒和细胞反应进行的。我们发现DV有效地感染了原发性人类EC，病毒滴度（〜105/ml）1天P.I.我们的发现表明，DV感染的EC有助于病毒血症，病毒传播以及对EC的DV抗原的呈现，使它们成为抗体和免疫细胞的靶标。我们对EC对DV感染反应的分析表明，直接免疫细胞募集和激活的趋化因子的高水平诱导。细胞因子CXCL10，CXCL11，IL-7，RANTES和BAFF分别诱导337、45、128、84和119倍，在同步感染的ECS中，在24 HRS P.I.中。 CXCL10/11和Rantes分泌募集并激活白细胞和T细胞，而IL-7刺激B，T和NK细胞增殖和T细胞成熟。 BAFF是在激活EC表面释放或表达的B和T细胞激活因子。 DV感染还诱导了适当的（因子P，34倍），它激活了产生C3A/C5A的替代途径（AP）补体系统：趋化的过敏毒素，趋于局部炎症，肥大细胞脱粒和血管渗透性。重要的是，严重病患者患者存在高水平的C3A，并且与严重疾病，血浆泄漏和休克有关。这些新发现的EC反应表明，EC不仅是DV感染的靶标，而且是释放因受感染内皮的免疫细胞趋化性和渗透性增强的因素。这些发现共同表明，DV感染的EC有助于病毒血症，增强的免疫反应和血管通透性，这是DHF和DSS的基本成分。我们建议定义对DV感染的EC反应，该反应通过激活和募集对内皮的免疫细胞来增强免疫反应。 公共卫生相关性：登革热病毒是由蚊子携带的，每年感染500-1亿人，导致500,000-1,000,000/年的血管泄漏疾病，死亡率为5-30％。登革热导致免疫增强的疾病过程，从而从独特的血清型中更严重。 DV感染了毛细管并调节水肿和出血的免疫细胞和内皮细胞。我们发现，DV感染的内皮细胞引起的反应增强了免疫介导的反应，从而有助于血管泄漏并有助于增加病毒和扩散。在这项研究中，我们在内皮细胞中解决了这一过程。