Genotypic and functional properties of HIV-1 Nef clinical isolates in PAH-HIV

PAH-HIV 中 HIV-1 Nef 临床分离株的基因型和功能特性

• 批准号: 8639346
• 负责人: TODD M BULL
• 金额: $ 66.43万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-26 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8639346
• 关键词: Address; Age; Alleles; Amino Acid Substitution; Amino Acids; Apoptosis; Biological Markers; Blood; Blood Banks; Brain; California; Cardiac Catheterization Procedures; Cell Proliferation; Cell physiology; Cells; Chronic; Clinical; Clone Cells; Coculture Techniques; Collaborations; Colorado; Complication; Data; Development; Down-Regulation; Echocardiography; Endothelial Cells; Environment; Evolution; Exercise; Exposure to; Frequencies; Functional disorder; Gene Expression; Genetic Polymorphism; Genetic Transcription; Genetic Variation; HIV; HIV-1; Human; Immune; Immune response; Immune system; In Vitro; Individual; Infection; Lung; Lung Lavage Fluid; Lung diseases; Lymphocyte Function; Macaca; Measures; Mitogens; Modeling; Molecular; Molecular Cloning; Molecular Profiling; National Heart, Lung, and Blood Institute; Pathogenesis; Patients; Peripheral Blood Mononuclear Cell; Phenotype; Plasma; Population; Positioning Attribute; Process; Property; Protein Kinase; Proteins; Pulmonary Hypertension; Pulmonary artery structure; Recombinants; Reporting; Research; Resistance; Rest; Risk; Role; Sampling; San Francisco; Scientist; Seminal fluid; Signal Transduction Pathway; Site; Specimen; Systolic Pressure; T cell response; T-Cell Receptor; T-Lymphocyte; Universities; Variant; Vascular Diseases; Vascular Endothelial Cell; Vascular remodeling; Viral; Viral Proteins; Virion; Virus; biobank; cell injury; cohort; design; hemodynamics; in vivo; nef Genes; nef Protein; novel; peripheral blood; pressure; public health relevance; pulmonary arterial hypertension; pulmonary artery endothelial cell; receptor density; receptor expression; receptor function; repository; response to injury; tool

DESCRIPTION (provided by applicant): As HIV-infected individuals continue to age, non-infectious complications increase in frequency. Despite the first descriptions of patient cases in 1987, little is known about the pathogenesis of pulmonary hypertension associated with HIV (PAH-HIV). Chronic exposure to viral products such as HIV-1 Nef and HIV-induced immune deregulation in the lung may contribute to pulmonary vascular disease, particularly through their impact on pulmonary endothelial cells (EC). Our group was the first to associate the HIV-1 nef protein with the pathogenesis of vascular remodeling PAH-HIV. We longitudinally followed 34 individuals with PAH-HIV and collected sequential clinical and echocardiographic; we created and curated a repository of plasma, bronchial lavage fluid and cells. We sequenced the nef gene from blood and lung samples from patients with PAH, elevated pulmonary artery systolic pressures and from non- PAH HIV infected. We found amino acid substitutions in the Nef protein statistically associated with the PAH phenotype; these substitutions clustered around Nef functional domains that potentially interfere with Nef adaptor functions. Further studies, which are preliminarily reported in this application, suggest that particular amino acid signatures predominate in the lungs compared with the periphery. The lung may be a protected environment that allows the virus to evade immune responses; furthermore, particular nef alleles will enhance T cell responses and result in pulmonary vascular endothelial cell proliferation/apoptosis, disrupt signal transduction pathways and result in vascular remodeling. We hypothesize that allelic variants of nef will have an impact on T cell and pulmonary endothelial cell function. We propose to 1) infect T cells with molecularly cloned virions containing primary nef alleles and measure T cell receptor density; 2) human pulmonary artery endothelial cells will be co-cultured with infected T cells or transfected with nef molecular clone and endothelial cell gene expression, apoptosis and proliferation measured and 3) determine whether the lung is a protected compartment for evolution of nef sequences. These studies will examine the functional properties of nef alleles containing these amino acid substitutions from PAH-HIV individuals. The studies proposed in this application will use existing biospecimens and cloned nef alleles to examine the mechanisms whereby HIV-nef influences pulmonary vascular remodeling in the pathogenesis of PAH-HIV. Our research tem, with a combination of basic and clinician scientists is well poised to address how the nef viral protein and immune dysregulation are contributing factors to this lung complication of HIV.

描述（由申请人提供）：随着艾滋病毒感染者继续年龄，非感染并发症的频率增加。尽管在1987年对患者病例进行了首次描述，但对与HIV相关的肺动脉高压的发病机理知之甚少（PAH-HIV）。长期暴露于病毒产物（例如HIV-1 NEF）和HIV诱导的肺部免疫管制可能会导致肺血管疾病，尤其是通过对肺内皮细胞（EC）的影响。我们的小组是第一个将HIV-1 NEF蛋白与血管重塑PAH-HIV的发病机理相关联的组。我们纵向跟踪了34个患有PAH-HIV的人，并收集了顺序的临床和超声心动图。我们创建并策划了血浆，支气管灌洗液和细胞的存储库。我们从血液和肺部样本中对NEF基因进行了对PAH患者，肺动脉收缩压升高以及未感染非PAH HIV的患者的NEF基因。我们在NEF蛋白中发现与PAH表型相关的氨基酸取代。这些取代聚集在NEF功能域周围，可能会干扰NEF适配器函数。在本应用中最初报道的进一步研究表明特定的氨基酸特征 与周围相比，肺部占主导地位。肺可能是一个受保护的环境，使病毒能够逃避免疫反应。此外，特定的NEF等位基因将增强T细胞反应并导致肺血管内皮细胞增殖/凋亡，破坏信号转导途径并导致血管重塑。我们假设NEF的等位基因变体将对T细胞和肺内皮细胞功能产生影响。我们建议1）用含有原发性NEF等位基因的分子克隆病毒体感染T细胞并测量T细胞受体密度； 2）人类肺动脉内皮细胞将与感染的T细胞共培养，或与NEF分子克隆和内皮细胞基因表达，凋亡和增殖测量的转染和3）确定肺是否是NEF序列进化的受保护的区域。这些研究将检查来自PAH-HIV个体的这些氨基酸取代的NEF等位基因的功能性能。 本应用中提出的研究将使用现有的生物测量和克隆的NEF等位基因来检查HIV-NEF在PAH-HIV的发病机理中影响肺血管重塑的机制。我们的研究TEM结合了基础和临床医生科学家的结合，可以很好地解决NEF病毒蛋白和免疫失调如何促成HIV肺并发症的因素。