Defining Inhibitory Mechanisms of Novel CCRS-targeted Microbicide Candidates

项目1-项目1的目标是确定三种新的趋化因子类似物的作用机制 具有阻断HIV-1通过CCR5进入的有效活性。这些完全重组的分子展示了三个 不同的活性谱：（组I）CCR5阻断而无信号传导活性或受体内化;（组II）CCR5阻断而无信号传导活性或受体内化。 II），具有信号传导活性的CCR5快速内化;和（组III），中度CCR5内化和 没有信号传导活性的阻断。这些新分子在以下方面具有显著的潜在优势： 安全和生产成本。超过当前[B]，p，n 这些杀微生物剂需要更详细地了解它们的作用机制。我们将 定义II类分子的胞内螯合途径，并与III类分子进行比较 分子。我们将研究一些假设，以解释在缺乏抗病毒药物的情况下延长抗病毒活性。 细胞内受体螯合（I组）或中度内化（III组），包括 CCR5二聚体形成、膜中受体定位改变、变构效应和受体 不依赖于G蛋白连接的信号转导的内化。我们还将研究CCR5，CCL5， CCL3L1基因多态性对CCR5蛋白合成和周转率的影响。这些研究 旨在研究来自正常人供体的原代靶细胞对 每一种新的抑制剂我们还将使用一组突变的CCR5分子来检查结构 活动的相关性。我们将利用这些关于机制和靶细胞变异性的信息来开发新的 分子具有更好的活性特征，我们将与其他项目进行协调实验， 在这个项目中，我们将在基于细胞的模型中的发现与当前和新的CCR5抑制剂的作用联系起来 在组织外植体和整个动物模型中。这种方法将产生更好、更安全的CCR5抑制剂， 能够以适合于在受影响最严重的地区阻止艾滋病毒/艾滋病蔓延的规模生产。

Project 1 - The goal of Project 1 is to define the mechanism of action of three new chemokine analogues with potent activity in blocking HIV-1 entry via CCR5. These fully recombinant molecules display three distinct activity profiles: (group I) CCR5 blockade without signaling activity or receptor internalization; (group II), CCR5 rapid internalization with signaling activity; and (group III), moderate CCR5 internalization and blockade without signaling activity. These new molecules have significant potential advantages in terms of safety and cost of production ..over currenynh[b^ fwither, irrtpr.pvejTients.,p,n these bahdidate'microbicides requires more detailed knowledge of their mechanisms of action. We will define the route to intracellular sequestration for group II molecules, and compare with that of the group III molecules. We will examine a number of hypotheses to explain prolonged antiviral activity in the absence of intracellular receptor sequestration (group I) or with moderate internalization (group III), including changes in CCR5 dimer formation, altered receptor localization in the membrane, allosteric effects, and receptor internalization independent of G-protein-linked signaling. We will also examine the impact of CCR5, CCL5, and CCL3L1 genetic polymorphisms on CCR5 protein synthesis and turnover rate. These studies are designed to investigate variability in the susceptibility of primary target cells from normal human donors to each of the new inhibitors. We will also use a panel of mutant CCR5 molecules to examine structural correlates of activity. We will use this information on mechanism and target cell variability to develop new molecules with even better activity profiles, and we will perform coordinated experiments with other projects in this Program to relate our findings in cell-based models to the effects of current and new CCR5 inhibitors in tissue explant and whole animal models. This approach will generate better and safer CCR5 inhibitors that can be produced on a scale suitable for stopping the spread of HIV/AIDS in the most-impacted areas.