New Model for NeuroAIDS Utilizing Monocyte-Derived HIV-1

描述(申请人提供)：我们进行了一项系统的研究，以评估各种抗逆转录病毒疗法(ART)方案对体内M/M中HIV-1复制的有效性，并确定M/M在抑制ART期间是否是HIV-1复制的来源。我们对HIV-1在血液单核细胞中的复制和遗传进化进行了纵向研究，并将这些结果与接受各种ART、间断ART或不接受ART的患者的CD4T细胞中的结果进行了比较。我们的结果表明：(A)循环单核细胞中的HIV-1在遗传上是异质性的，反映了HIV-1与CD4T细胞中的HIV-1不同的遗传特征；(B)在HIV-1感染过程中，无论是否有ART，HIV-1在M/M中作为独立于CD4T细胞的一个隔室不断复制和进化；(C)ART期间的病毒抑制减少了T细胞内HIV-1序列的进化，而单核细胞内的HIV-1继续进化；(D)HAART治疗期间从单核细胞获得的HIV-1与T细胞内的病毒相比，对抗逆转录病毒药物的敏感性可能降低；以及(E)在单核细胞内存在一种独特的、但之前未描述的HIV-1表型，它支持CCR5介导的巨噬细胞感染，但不支持T细胞感染。单核细胞来源的HIV-1和单核细胞来源的巨噬细胞选择性R5(MDMS-R5)组病毒的不同基因型和表型的发现表明，在血液单核细胞中循环的HIV-1代表复制的HIV-1，这可能是由不同组织中的巨噬细胞产生的，而不同组织的巨噬细胞可能不受当前ART方案的抑制。然而，所有上述基因型和表型结果都是从患者纯化的单核细胞DNA中提取的PCR扩增的HIV-1 DNA获得的。目前尚不清楚这些前病毒是否真的会复制并产生传染性病毒。该项目将是建立和表征从患者纯化的M/M中分离出的一组新的传染性HIV-1的第一步。然后，我们将确定“天然”M/M-HIV-1与M/M系细胞的相互作用，包括与中枢神经系统相关/衍生的细胞类型。最后，我们将利用这一组天然的M/M来源的HIV-1在体外和/或体外模型中评估药物的抗HIV活性，旨在开发针对M/M中HIV-1感染的新治疗药物，并防止HIV-1感染的巨噬细胞对中枢神经系统的损伤。项目简介：我们将建立和描述从M/M分离的HIV-1的一个新的小组，并定义单核细胞来源的HIV-1与中枢神经系统细胞的相互作用。此外，我们还将在体外模型中利用这一组天然的M/M来源的HIV-1来评估目前批准的药物和新药的抗HIV活性，旨在开发新的治疗药物，防止这些单核细胞来源的HIV-1感染的巨噬细胞引起的中枢神经系统损伤。

DESCRIPTION (provided by applicant): We have performed a systematic study to evaluate the effectiveness of a variety of antiretroviral therapy (ART) regimens on HIV-1 replication in M/M in vivo, and determined whether M/M are sources of HIV-1 replication during suppressive ART. We have longitudinally studied HIV-1 replication and genetic evolution in blood monocytes, comparing these results with those in CD4+ T cells from patients who received a variety of regimens of ART, discontinuous ART, or no ART. Our results indicate that (a) HIV-1 contained within circulating monocytes are genetically heterogeneous and reflect genetically distinct HIV-1 from those in CD4+ T cells; (b) HIV-1 continuously replicates and evolves in M/M as a separate compartment from CD4+ T cells during the course of HIV-1 infection with or without ART; (c) virus suppression during ART decreased HIV-1 sequence evolution within T cells, while continued evolution in monocytes occurs; (d) HIV-1 obtained from monocytes during HAART therapy may have a decreased sensitivity to antiretroviral drugs compared with virus within the T cell compartment; and (e) there exists a unique, but not previously-described HIV-1 phenotype within the monocyte compartment, that supports CCR5-mediated infection of macrophages but not T cells. Findings of heterogeneous genotypes and phenotypes of monocyte-derived HIV-1 and the monocyte-derived macrophage-selective R5 (MDMS-R5) groups of viruses suggest that HIV-1 circulating in blood monocytes represent replicating HIV-1 which may be produced from macrophages in different tissues less suppressed by current regimens of ART. However, all above genotype and phenotype results were obtained from PCR-amplified HIV-1 DNA, derived from the patients' purified monocyte DNA. Whether these proviruses actually replicate and produce infectious virus remains unknown. This project will be the first step in establishing and characterizing a novel panel of infectious HIV-1 that will be isolated from patients' purified M/M. We will then define the interaction of "natural" M/M-HIV-1 with M/M lineage cells including CNS- related/derived cell types. Finally we will evaluate the anti-HIV activity of drugs in ex vivo and/or in vitro models using this panel of "natural" M/M-derived HIV-1, aiming to develop new therapeutic agents targeting HIV-1 infection in M/M, and preventing CNS damage caused by HIV-1 infected macrophages. Project Narrative: We will establish and characterize a novel panel of HIV-1 isolated from M/M, and define the interaction of monocyte-derived HIV-1 with CNS cells. In addition, we will evaluate the anti-HIV activity of currently approved and new drugs in ex vivo models by using this panel of "natural" M/M-derived HIV-1, aiming to develop new therapeutic agents preventing CNS damage caused by these monocyte-derived HIV-1 infected macrophages.