HIV and CD16+ monocytes as cellular reservoir & HIV/pulmonary hypertension

HIV 和 CD16 单核细胞作为细胞库

• 批准号: 8269667
• 负责人: SHARILYN ALMODOVAR
• 金额: $ 13.61万
• 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/8269667
• 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; angiogenesis; base; cell injury; cellular imaging; chemokine; cytokine; extracellular; ikarugamycin; in vivo; inhibitor/antagonist; injured; macrophage; magnetic beads; migration; monocyte; mortality; nef Genes; nef Protein; normotensive; peripheral blood; pressure; receptor expression; simian human immunodeficiency virus; success; trafficking

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.

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然知之甚少。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。在感染的急性期，HIV-1在外周血液和末梢器官中广泛复制。在血清转换和HAART抑制HIV病毒载量后，患者可能会经历一段亚临床病毒血症阶段(HIV残留复制)。这一阶段是指病毒在低于检测水平的情况下复制的阶段。在这一阶段，组织可能会补种一些在外周血浆中不一定检测到的准种。几个解剖部位为HIV提供了病毒复制和进化的保护性隔间；这些可能是亚临床病毒血症的重要来源。这些隔室包括脑、肠道相关淋巴组织、骨髓和生殖道等。目前的研究重点是在HIV残留复制过程中寻找淋巴细胞以外的细胞库。有证据表明，髓系细胞是病毒的细胞库，而不是血浆病毒的补种。与普通人群相比，HIV+患者更容易被诊断为HIV相关性肺动脉高压(HRPAH)，而不考虑抑制性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。我们的团队已经证明，HIV Nef与感染Shiv-nef的猕猴肺部的血管重塑和管腔闭塞的PAH样病变有关。值得注意的是，这些病变只出现在肺部(而不是其他器官)，并且在此之前有炎性浸润物。其他研究也表明，HIV包膜糖蛋白120也可以诱导血管重塑。此外，我们有证据表明，被诊断为真正的HRPAH的患者在外周血中含有特定的nef等位基因。目前尚不清楚HRPAH患者是否只有一个标志性的Nef，还是具有env等基因协同进化的标志性HIV准种。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仍有待阐明。申请人和其他人已经证明，经典的CD14+单核细胞在抑制HAART期间不携带HIV，外周血中的CD16+促炎单核细胞含有不同于病毒症患者血浆中的病毒准种。外周血CD16+单核细胞类似于肺泡巨噬细胞，感染HIV后可通过内皮迁移。鉴于这些有趣的观察，我们认为HRPAH患者与HRPAH相关的准种。HIV感染者外周血中的CD16+单核细胞是针对这些特定的HIV准种的特洛伊木马。在到达肺并激活后，这些促炎症的肺单核/巨噬细胞向肺内重新接种病毒粒子，开始恶性循环，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。我们将使用几种实验室技术-单基因组扩增、系统发育分析、使用磁珠分离细胞、国际电信联盟杂交中的荧光和体内细胞成像。