Mechanism of HIV-1 inhibition by alpha-defensin-1

α-defensin-1 抑制 HIV-1 的机制

• 批准号: 7054394
• 负责人: Theresa L Chang
• 金额: $ 29.66万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-06-01 至 2006-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7054394
• 关键词: AIDS therapy; antiAIDS agent; antibiotics; antiviral agents; biological signal transduction; chemical structure function; defensins; drug discovery /isolation; enzyme activity; enzyme inhibitors; helper T lymphocyte; human immunodeficiency virus 1; immunopharmacology; intermolecular interaction; life cycle; macrophage; microorganism disease chemotherapy; peptide structure; protein kinase C; therapy design /development; virion; virus replication

The overall objective of this proposal is to understand the mechanism by which a-defensin-1 inhibits HIV-1 replication. Alpha-defensins are cationic, cysteine (Cys)-rich peptides and play an important role in innate host defense. Recent studies including ours clearly demonstrate that alpha-defensin-1 has anti-HIV-1 activity. Our recent finding demonstrates that adefensin- 1 acts on the virion and the cell, although the direct effect on virions occurs under restricted conditions. We have shown that alpha-defensin-1 at a non-cytotoxic, physiological concentration can inhibit HIV-1 replication following viral entry, suggesting that alpha-defensin-1 has an effect(s) on the target cells where HIV-1 replicates. In primary CD4+ T cells, we have identified that alpha-defensin-1 inhibits protein Kinase C (PKC) signaling, a pathway important for HIV-1 infection and immune cell function. Our hypothesis is that alpha-defensin-1 has significant anti-HIV-1 activity and this activity is largely mediated through cell signaling in target cells. This proposal will study the mechanism(s) of action in primary CD4+ T cells and macrophages, the major target cells for HIV-1. In Aim 1, we will extend the post-entry study by evaluating the antiviral activity of alpha-defensin-1 against HIV-1 primary isolates. The direct effect on HIV-1 virions will be further studied using R5 viruses. We will identify the point in the virus life cycle blocked by a-defensin-1 in these primary cells. In Aim 2, we will assess the role of PKC in alpha-defensin- 1-mediated HIV-1 inhibition. Our preliminary results indicate that PKCalpha and (3 are involved in HIV-1 inhibition by alpha-defensin-1. We will assess activities of PKCalpha and (3 in primary CD4+ T cells in response to a-defensin-1. We will also evaluate the interaction of alpha-defensin-1 with PKCalpha or beta. The role of PKCalpha and beta in alpha-defensin-1-mediated anti-HIV activity will be examined by introducing constitutively active PKC. In Aim 3, we will study the contribution of structure of a-defensin-1 to the anti-HIV-1 activity. We will explore the difference in native compared to recombinant and synthetic alpha-defensin-1 in anti-HIV affect. We will define amino acid residues in a-defensin-1 important for its anti-HIV-1 activity. These studies will offer insights into the function of alpha-defensin-1 in innate immune responses against HIV-1 infection and provide an understanding of the structure-function relationship of a-defensin-1 that is essentially important in view of the efforts to develop defensin-derived peptides for prevention and therapeutic use.

该提案的总体目的是了解A-Defensin-1抑制HIV-1复制的机制。 α-防御素是阳离子，半胱氨酸（CYS） - 富含肽，在先天宿主的防御中起着重要作用。最近的研究包括我们的α-防御素1具有抗HIV-1活性。我们最近的发现表明，adefensin- 1对病毒粒子和细胞作用，尽管对病毒体的直接影响发生在受限之下 状况。我们已经表明，在非周期毒性，生理浓度下的α-二防御素-1可以抑制病毒进入后的HIV-1复制，这表明α-德法素-1对HIV-1复制的靶细胞具有影响（S）。在原代CD4+ T细胞中，我们已经鉴定出α-防御素-1抑制蛋白激酶C（PKC）信号传导，这是对HIV-1感染和免疫细胞功能重要的途径。我们的假设是α-防fefensin-1具有显着的抗HIV-1活性，并且该活性在很大程度上通过靶细胞中的细胞信号传导介导。该建议将研究原代CD4+ T细胞和巨噬细胞（HIV-1的主要靶细胞）中作用机理。在AIM 1中，我们将通过评估Alpha-Defensin-1对HIV-1主要分离株的抗病毒活性来扩展进入后研究。将使用R5病毒进一步研究对HIV-1病毒体的直接影响。我们将确定这些原代细胞中A-二防御素-1阻断的病毒生命周期中的点。在AIM 2中，我们将评估PKC在α-防御素1介导的HIV-1抑制中的作用。我们的初步结果表明，pkcalpha和（3涉及α-防御素1的HIV-1抑制作用。我们将评估PKCALPHA的活性和（响应于A-Defensin-1的主要CD4+ T细胞中的3个。我们还将评估Alpha-Defensin-1与PKCALPHA的相互作用。 α-Defensin-1介导的抗HIV活性将通过在AIM 3中引入组成型PKC，我们将研究A-Defensin-1对抗HIV-1活性的贡献。活动。