Innate Immunity, HIV, and OIs: Role of TLRs in the Lung

先天免疫、HIV 和 OIs：TLR 在肺中的作用

• 批准号: 6735646
• 负责人: Paul R Skolnik
• 金额: $ 22.83万
• 依托单位: BOSTON MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-05-01 至 2005-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6735646
• 关键词: HIV envelope protein; HIV infections; alveolar macrophages; biological signal transduction; clinical research; enzyme linked immunosorbent assay; flow cytometry; host organism interaction; human subject; immunity; immunocytochemistry; lipopolysaccharides; lung; microorganism immunology; nuclear factor kappa beta; opportunistic infections; patient oriented research; peptidoglycan; protein protein interaction; protein quantitation /detection; receptor binding; receptor expression; toll like receptor; transfection /expression vector; tumor necrosis factor alpha; western blottings

Opportunistic infections (OIs) in patients with AIDS often occur in the lung. These lead to substantial morbidity and mortality, even in the era of highly-active antiretroviral therapy (HAART). The innate immune system has been recognized as an important component of host immunity to infectious pathogens, but these responses have received little attention in the setting of HW infection, since most studies have focused on the adaptive immune system. We propose to study alterations in Toll-like receptors (TLRs), a central component of innate immune responses, during HIV infection. These studies will use patient-derived samples from the lung (alveolar macrophages, AMs) to most closely mimic the in vivo situation, and monoblastoid cell lines (U1 and U937 cells) to model the patient-derived cells and study the detailed mechanisms of these alterations. The effects of HIV in the lung and subsequent functional alterations of immune effector cells in the lung, are understudied areas. Our central hypothesis is that HIV infection causes alterations in TLR expression that lead to deficient innate immune responses, which predispose patients to OIs. We have preliminary data showing decreased TLR2 and TLR4 mRNA and protein expression, and decreased TNFcx levels (a TLR-mediated effector function) in AMs from HIV+ subjects compared to HIV- controls. We will study whether TLRs are altered during HIV infection, in AMs from HIV+ and HIV- subjects, and in U1 and U937 cells, using relevant TLR2 and TLR4 hgands, by determining (i) TLR2 (and TLR1 and TLR6, which form heterodimers with TLR2) and TLR4 mRNA and protein expression; (ii) TLR2- and TLR4-mediated TNFc_ production; (iii) phosphorylated signaling intermediaries of the TLR pathway (MyD88, IRAK, and PI3'-kinase), and concomitant NF-kB DNA-binding activity; and (iv) if HW-1 gp120 or gp160 affect TLR2 or TLR4 expression, either by directly binding to these TLR, or through indirect effects. The innovation of this grant is a new paradigm that suggests direct or indirect effects of HIV on TLRs that may affect innate immune responses to OIs or HIV, itself.

艾滋病患者的肺部感染（OIs）通常发生在肺部。即使在高效抗逆转录病毒疗法（HAART）时代，这些也会导致严重的发病率和死亡率。先天免疫系统已被认为是宿主对感染性病原体免疫的重要组成部分，但这些反应在HW感染的情况下很少受到关注，因为大多数研究都集中在适应性免疫系统上。我们建议研究Toll样受体（TLR）的变化，先天免疫反应的核心组成部分，在HIV感染。这些研究将使用来自肺的患者源性样本（肺泡巨噬细胞，AM）来最接近地模拟体内情况，并使用单核细胞系（U1和U937细胞）来模拟患者源性细胞并研究这些改变的详细机制。HIV在肺中的作用和随后肺中免疫效应细胞的功能改变是研究不足的领域。我们的中心假设是，HIV感染导致TLR表达的改变，导致先天免疫反应不足，使患者易患OI。我们的初步数据显示，与HIV-对照相比，HIV+受试者的AM中TLR 2和TLR 4 mRNA和蛋白表达降低，TNFcx水平降低（TLR介导的效应功能）。我们将研究TLR在HIV感染过程中，在HIV+和HIV-受试者的AM中，以及在U1和U937细胞中是否发生改变，使用 相关的TLR2和TLR4表达，通过测定（i）TLR2（和TLR1和TLR6，其与TLR2形成异二聚体）和TLR4 mRNA和蛋白质表达;（ii）TLR2和TLR4介导的TNFc_产生;（iii）TLR途径的磷酸化信号传导中间体（MyD88、IRAK和PI3 ′-激酶）和伴随的NF-κ B DNA结合活性;和（iv）HW-1 gp120或gp160是否通过直接结合这些TLR或通过间接作用影响TLR 2或TLR 4表达。这项赠款的创新是一种新的范式， 或HIV对TLR的间接影响，其可能影响对OI或HIV本身的先天免疫应答。