4R CEMBRANOID

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 多年来，艾滋病毒/艾滋病的治疗已扩大到包括26种药物。然而，抗药性病毒株的快速发展使抗逆转录病毒治疗在某些情况下暂时有效，在另一些情况下，稳定静止的CD4T细胞作为潜伏感染的蓄水池的概念代表了治愈感染的主要障碍。目前使用的高效抗逆转录病毒疗法(HAART)一般都很昂贵，毒性相对较大，生物利用度很低，特别是对脑组织。我们了解到有一个重大的 需要确定一种化合物，可以预防或减轻艾滋病毒-L感染后的破坏性影响。仍然需要继续努力发现新的抗艾滋病毒药物，特别是具有新的抗艾滋病毒机制的新小分子药物。 我们已经确定了一种名为4R的无毒西膜类化合物，它很容易渗透到大脑中，具有强大的抗艾滋病毒和抗炎活性。因此，这项建议旨在测试4R在体内抑制艾滋病毒-L复制的能力 从艾滋病毒感染者身上提取淋巴细胞，并调节感染细胞中抗炎细胞因子和趋化因子的产生。为了了解这些活动的机制，我们将检验以下假设： 新化合物4R通过与α7烟碱型乙酰胆碱受体相互作用抑制HIV-L的复制和炎症，从而调节抗病毒Wnt/β-catenin和抗炎Stan通路。这个 以下三个具体目标被提出来检验这一假设，使用来自20名HIV阴性的健康志愿者和30名HIV阳性患者的PBMC(来自HIV阴性的人的PBMC将急性感染HIV-1SF2(X4/RS)双重病毒 菌株)。目的1.应用HIV-L逆转录聚合酶链式反应(RT-PCR)检测4R能否抑制HIV感染者外周血单核细胞中HIV的复制。目的2.确定4R抗HIV活性是否由HIV复制相关的调控所介导 细胞因子和趋化因子的细胞学珠阵(CBA)分析。目的3.确定4R抗HIV和抗炎活性是否分别通过激活Wnt/β-catenin和Stan通路而介导。 蛋白质印迹分析。此外，为了测试4R活性是否源于与α7烟碱型乙酰胆碱受体(A7nAchR)的相互作用以及随后对细胞内钙水平的调节，将进行钙成像。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The therapy of HIV/AIDS has expanded over the years to include 26 drugs. However, rapid development of drug resistantviral strains makes antiretroviral therapy for HIV infection transiently effective in some scenarios, in others the concept of stable quiescent CD4 T cell functioning as reservoirs of the latent infection represent the mayor hurdle towards the cure of the infection. The currently used regimens of Highly Active Antiretroviral Therapy (HAART) are generally expensive, relatively toxic and carry an inefficient bioavailability particularly to the brain tissue. We understand that there is a major need to identify a compound that can prevent or alleviate the damaging effects following HIV-l infection. Continued effort for the discovery of new anti-HIV drugs, especially for new small molecules with novel anti-HIV mechanisms, is still needed. We have identified a non-toxic cembranoid called 4R that readily penetrates into the brain and has both potent anti-HIV and anti-inflammatory activity. Therefore, this proposal aims to test 4R for its ability to suppress HIV-l replication in lymphocytes from HIV-infected individuals and to modulate the production of anti-inflammatory cytokines and chemokines in the infected cells. In an attempt to understand the mechanism of these activities, we will test the following hypothesis: the novel compound 4R inhibits HIV-l replication and inflammation by interacting with the alpha7 nicotinic acetylcholine receptor which results in modulation of the antiviral Wnt/beta-catenin and the anti-inflammatory STAn pathways. The following three specific aims are proposed to test this hypothesis, using PBMC from 20 HIV-negative healthy volunteers and 30 HIV-positive patients (PBMC from HIV-negative individual will be acutely infected with HIV-1SF2 (X4/RS) dual virus strain). Aim 1. To determine whether 4R can suppress HIV replication in PBMC from HIV-infected patients using HIV-l RT PCR. Aim 2. To determine whether 4R anti-HIV activity is mediated by the modulation of HIV replication-associated cytokines and chemokines using cytometric bead array (CBA). Aim 3. To determine whether 4R anti-HIV and antiinflammatory activities are mediated by the activation of the Wnt/beta-Catenin and STAn pathways, respectively using Western blot analysis. In addition, to test whether 4R activities are due to interaction with the alpha7 nicotinic acetylcholine receptor (a7nAchR) and subsequent modulation of the intracellular calcium levels, calcium imaging will be performed.