HIV evolution and CD16+ monocytes as cellular reservoirs: insights into HIV/pulmo

HIV 进化和 CD16 单核细胞作为细胞库：对 HIV/pulmo 的见解

• 批准号: 7923064
• 负责人: SHARILYN ALMODOVAR
• 金额: $ 13.63万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-16 至 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7923064
• 关键词: AIDS/HIV problem; Acute; Affect; Alleles; Alveolar Macrophages; Amino Acid Sequence; Antibodies; Award; Biological; Blood; Blood specimen; Bone Marrow; Brain; Bronchoalveolar Lavage; Bronchoalveolar Lavage Fluid; Bronchoscopy; CD14 gene; CD3 Antigens; CD4 Antigens; California; Cardiac Catheterization Procedures; Cell Separation; Cells; Cellular Membrane; Clinical; Coupled; Detection; Diagnosis; Disease; Down-Regulation; Endothelial Cells; Engineering; Enrollment; Equus caballus; Evolution; Exhibits; FCGR3B gene; Fingerprint; Flow Cytometry; Flowcharts; Fluorescent in Situ Hybridization; Funding; General Population; Genes; Genital system; Genome; Genomics; Glycoproteins; Gut associated lymphoid tissue; HIV; HIV diagnosis; HIV-1; Highly Active Antiretroviral Therapy; Human; Hydrolase; In Vitro; Individual; Infection; Inflammation; Inflammatory; Inflammatory Infiltrate; Laboratories; Lesion; Life; Ligands; Link; Lung; Lymphocyte; Lymphoid; Macaca; Measurement; Membrane Microdomains; Mentors; Mitogen-Activated Protein Kinases; Molecular; Myelogenous; Myeloid Cells; National Heart, Lung, and Blood Institute; Organ; Pathogenesis; Pathway interactions; Patients; Peptide Sequence Determination; Peripheral; Peripheral Blood Mononuclear Cell; Phase; Phenotype; Phylogenetic Analysis; Plasma; Population; Production; Proteins; Pulmonary Hypertension; Pulmonary artery structure; Recombinants; Research; Residual state; Rest; Role; Sampling; San Francisco; Sequence Analysis; Signal Transduction; Site; Source; Specimen; Staging; T-Lymphocyte; Techniques; Testing; Time; Tissues; Translating; Tropism; Universities; Variant; Vascular Endothelial Cell; Vascular Endothelium; Vascular remodeling; Viral; Viral Load result; Viremia; Virion; Virus; Work; abstracting; angiogenesis; base; cell injury; cellular imaging; chemokine; cytokine; extracellular; ikarugamycin; in vivo; inhibitor/antagonist; injured; insight; macrophage; magnetic beads; migration; monocyte; mortality; nef Genes; nef Protein; normotensive; peripheral blood; pressure; public health relevance; receptor expression; success; trafficking

DESCRIPTION (provided by applicant): The dynamics of viral replication and evolution in the lungs of patients infected with Human Immunodeficiency Virus type-1 (HIV-1) remain poorly understood. In addition, pulmonary complications are still prevalent despite the unquestioned success of Highly Active Antiretroviral Therapy (HAART). HIV-1 replicates extensively during the acute phase of infection in peripheral blood and end organs. After seroconversion, and suppression of HIV viral loads by HAART, patients may undergo a phase of sub-clinical viremia (HIV residual replication). This phase refers to a stage in which the virus replicates below the levels of detection. During this phase, tissues may be reseeded with quasispecies that are not necessarily detected in the peripheral plasma. Several anatomical sites provide HIV with protected compartments for viral replication and evolution; these may be important sources for the sub-clinical viremia. These compartments include the brain, gut-associated lymphoid tissue, bone marrow, and genital tract, etc. Research efforts are now focused on the search for cellular reservoirs other than lymphocytes during the residual replication of HIV. Evidence points to myeloid cells as the cellular reservoirs of virus not reseeded from plasma viruses. HIV+ patients are more frequently diagnosed with HIV-related pulmonary hypertension (HRPAH) compared to the general population, regardless of suppressive HAART. HRPAH is a disease with a prominent inflammatory component that is also linked to higher mortality. Our group has demonstrated that HIV Nef is associated with vascular remodeling and lumen- obliterative PAH-like lesions in the lungs of SHIV-nef-infected macaques. Of note, these lesions appeared only in the lungs (not in other organs) and were preceded by inflammatory infiltrates. Others have also shown that HIV Envelope (Env) glycoprotein 120 induces vascular remodeling as well. Furthermore, we have evidence that patients diagnosed with bona fide HRPAH harbor specific nef alleles in the peripheral blood. Whether HRPAH patients harbor only a signature Nef or signature HIV quasispecies with genes like env evolving in concert remains unknown at this time. The cellular sources of HIV and their impact in the lung vasculature of HRPAH patients receiving suppressive HAART also remain to be elucidated. The applicant and others have shown that classic CD14+ monocytes do not carry HIV during suppressive HAART and CD16+ pro- inflammatory monocytes in peripheral blood harbor viral quasispecies different from those in plasma of viremic patients. Peripheral blood CD16+ monocytes resemble alveolar macrophages, and undergo transendothelial migration when infected with HIV. Given these intriguing observations, we think that HRPAH patients HRPAH- associated quasispecies. Peripheral blood CD16+ monocytes in HIV-infected patients serve as Trojan horses for these specific HIV quasispecies. After reaching the lung and upon activation, these pro-inflammatory lung monocyte/macrophages reseed the lung with such virions, starting a vicious cycle in which encoded Nef and Env increase the production of inflammatory molecules that may impact endothelial cells. Subsequently, injured endothelial cells may switch to a proliferative-angiogenic phenotype leading to the clinical picture of HRPAH. We will use several laboratory techniques single-genome amplification, phylogenetic analyses, cell isolation using magnetic beads, fluorescent in itu hybridization and in vivo cell imaging. PUBLIC HEALTH RELEVANCE: This project seeks to fingerprint the HIV in the lungs of infected patients and to discover how HIV proteins insults the endothelial cells in the lung vasculature. (End of Abstract)

描述（由申请方提供）：对人类免疫缺陷病毒1型（HIV-1）感染患者肺部病毒复制和进化的动力学仍知之甚少。此外，尽管高效抗逆转录病毒疗法（HAART）取得了毋庸置疑的成功，但肺部并发症仍然普遍存在。HIV-1在外周血和终末器官感染的急性期广泛复制。在血清转换和HAART抑制HIV病毒载量后，患者可能会经历亚临床病毒血症（HIV残留复制）阶段。这一阶段是指病毒复制低于检测水平的阶段。在此阶段，组织可能会被准种重新吸收，而这些准种不一定在外周血浆中检测到。几个解剖部位为HIV提供了病毒复制和进化的保护区室;这些可能是亚临床病毒血症的重要来源。这些隔间包括大脑，肠道相关的淋巴组织，骨髓，生殖道等研究工作现在集中在寻找细胞水库以外的其他淋巴细胞在剩余的复制艾滋病毒。有证据表明，骨髓细胞是病毒的细胞储存库，而不是从血浆病毒中回收。与一般人群相比，HIV+患者更常被诊断为HIV相关肺动脉高压（HRPAH），无论是否使用抑制性HAART。HRPAH是一种具有显著炎症成分的疾病，也与较高的死亡率有关。我们的小组已经证明，HIV Nef与SHIV-nef感染的猕猴肺中的血管重塑和管腔闭塞性PAH样病变相关。值得注意的是，这些病变仅出现在肺中（未出现在其他器官中），并且之前有炎性浸润。其他人也表明HIV包膜（Env）糖蛋白120也诱导血管重塑。此外，我们有证据表明被诊断患有真正的HRPAH的患者外周血中含有特定的nef等位基因。HRPAH患者是否只携带一个标志性Nef或标志性HIV准种，与env等基因协同进化，目前尚不清楚。HIV的细胞来源及其在接受抑制性HAART的HRPAH患者的肺血管系统中的影响仍有待阐明。申请人和其他人已经表明，经典的CD 14+单核细胞在抑制性HAART期间不携带HIV，并且外周血中的CD 16+促炎单核细胞携带与病毒血症患者血浆中的那些不同的病毒准种。外周血CD 16+单核细胞类似于肺泡巨噬细胞，在感染HIV时进行跨内皮迁移。鉴于这些有趣的观察结果，我们认为高血压肺动脉高压患者是高血压肺动脉高压相关的准种. HIV感染者外周血CD 16+单核细胞可作为这些特定HIV准种的特洛伊木马。在到达肺后并在激活时，这些促炎性肺单核细胞/巨噬细胞用这种病毒粒子重新接种肺，开始恶性循环，其中编码的Nef和Env增加可能影响内皮细胞的炎性分子的产生。随后，受损的内皮细胞可能转变为增殖性血管生成表型，导致HRPAH的临床表现。我们将使用几种实验室技术单基因组扩增，系统发育分析，细胞分离使用磁珠，荧光在国际电联杂交和在体内细胞成像。 公共卫生关系：该项目旨在对感染患者肺部的HIV进行指纹识别，并发现HIV蛋白如何损害肺血管系统中的内皮细胞。(End摘要）