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: 8128601
• 负责人: SHARILYN ALMODOVAR
• 金额: $ 13.66万
• 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/8128601
• 关键词: AIDS/HIV problem; Acute; Affect; Alleles; Alveolar Macrophages; Amino Acid Sequence; Antibodies; 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; 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; Infection; Inflammation; Inflammatory; Inflammatory Infiltrate; Laboratories; Lesion; Life; Ligands; Link; Lung; Lymphocyte; 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; simian human immunodeficiency virus; 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（由申请人提供）：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态仍知之甚少。此外，尽管高效抗逆转录病毒疗法（HAART）取得了无可争议的成功，但肺部并发症仍然普遍存在。 HIV-1 在感染急性期在外周血和终末器官中广泛复制。在血清转化和通过HAART抑制HIV病毒载量后，患者可能会经历一个亚临床病毒血症阶段（HIV残留复制）。该阶段是指病毒复制低于检测水平的阶段。在此阶段，组织可能会重新播种外周血浆中不一定检测到的准种。一些解剖部位为 HIV 病毒复制和进化提供了受保护的区室；这些可能是亚临床病毒血症的重要来源。这些区室包括大脑、肠道相关淋巴组织、骨髓和生殖道等。目前的研究工作重点是在艾滋病毒残留复制过程中寻找淋巴细胞以外的细胞储存库。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重新接种的。与普通人群相比，无论是否采用抑制性 HAART，HIV+ 患者更常被诊断出患有 HIV 相关肺动脉高压 (HRPAH)。 HRPAH 是一种具有显着炎症成分的疾病，也与较高的死亡率有关。我们的研究小组已经证明，HIV Nef 与 SHIV-nef 感染的猕猴肺部的血管重塑和管腔闭塞性 PAH 样病变有关。值得注意的是，这些病变仅出现在肺部（不在其他器官中），并且之前有炎症浸润。其他研究还表明，HIV 包膜 (Env) 糖蛋白 120 也会诱导血管重塑。此外，我们有证据表明，诊断为真正的 HRPAH 的患者外周血中含有特定的 nef 等位基因。 HRPAH 患者是否仅携带标志性 Nef 或标志性 HIV 准种（与 env 等基因一致进化）目前仍不得而知。 HIV 的细胞来源及其对接受抑制性 HAART 的 HRPAH 患者肺血管系统的影响也有待阐明。申请人和其他人已经证明，经典的CD14+单核细胞在抑制性HAART期间不携带HIV，并且外周血中的CD16+促炎单核细胞含有与病毒血症患者血浆中的病毒准种不同的病毒准种。外周血 CD16+ 单核细胞类似于肺泡巨噬细胞，当感染 HIV 时会进行跨内皮迁移。鉴于这些有趣的观察结果，我们认为 HRPAH 患者是 HRPAH 相关的准种。 HIV感染者的外周血CD16+单核细胞充当这些特定HIV准种的特洛伊木马。到达肺部并激活后，这些促炎性肺部单核细胞/巨噬细胞在肺部重新播种此类病毒粒子，开始恶性循环，其中编码的 Nef 和 Env 增加了可能影响内皮细胞的炎症分子的产生。随后，受损的内皮细胞可能转变为增殖血管生成表型，从而导致 HRPAH 的临床表现。我们将使用多种实验室技术，包括单基因组扩增、系统发育分析、磁珠细胞分离、荧光原位杂交和体内细胞成像。 公共健康相关性：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。 （摘要完）