New Model for NeuroAIDS Utilizing Monocyte-Derived HIV-1

描述（由申请人提供）：我们进行了一项系统的研究，以评估各种抗逆转录病毒治疗（ART）方案对M/M体内HIV-1复制的有效性，并确定M/M是否是抑制性ART期间HIV-1复制的来源。我们对血液单核细胞中的HIV-1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抗逆转录病毒治疗方案、间断抗逆转录病毒治疗方案或未接受抗逆转录病毒治疗的患者的CD4+ T细胞中的结果进行了比较。我们的研究结果表明(a)循环单核细胞中含有的HIV-1在遗传上是异质的，反映了与CD4+ T细胞中不同的HIV-1基因；(b)在接受或不接受抗逆转录病毒治疗的HIV-1感染过程中，HIV-1在M/M中作为一个独立于CD4+ T细胞的细胞室不断复制和进化；(c)抗逆转录病毒治疗期间的病毒抑制降低了T细胞内HIV-1序列的进化，而单核细胞则继续进化；(d)与T细胞室内的病毒相比，在HAART治疗期间从单核细胞获得的HIV-1可能对抗逆转录病毒药物的敏感性降低；(e)在单核细胞室中存在一种独特的，但以前没有描述过的HIV-1表型，它支持ccr5介导的巨噬细胞感染，而不是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）病毒群的基因型和表型异质性的研究结果表明，血液单核细胞中循环的HIV-1代表了复制的HIV-1，这可能是由不同组织的巨噬细胞产生的，而当前的抗逆转录病毒治疗方案对巨噬细胞的抑制较少。然而，上述所有基因型和表型结果均来自于pcr扩增的HIV-1 DNA，来源于患者纯化的单核细胞DNA。这些原病毒是否真的复制并产生传染性病毒仍然未知。该项目将是建立和描述从患者纯化的M/M中分离出的新型传染性HIV-1小组的第一步。然后，我们将定义“天然”M/M- hiv -1与M/M谱系细胞（包括中枢神经系统相关/衍生细胞类型）的相互作用。最后，我们将利用这组“天然”M/M衍生的HIV-1在离体和/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感染的新的治疗药物，并预防HIV-1感染的巨噬细胞引起的中枢神经系统损伤。项目描述：我们将建立并表征从M/M中分离的一组新的HIV-1，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，我们将利用这组“天然”M/M源性HIV-1在离体模型中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来预防这些单核细胞源性HIV-1感染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.