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治疗（ART）方案对体内M/M中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。我们纵向研究了血液单核细胞中HIV-1的复制和遗传进化，将这些结果与接受各种ART方案、间断ART或未接受ART的患者的CD 4 + T细胞中的结果进行比较。我们的结果表明：（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CD 4 + T细胞中的HIV-1在遗传上不同;（B）在有或没有ART的HIV-1感染过程中，HIV-1在M/M中作为与CD 4 + T细胞分开的区室连续复制和进化;（c）ART期间的病毒抑制减少了T细胞内的HIV-1序列进化，而在单核细胞中发生持续进化;（d）在HAART治疗期间从单核细胞获得的HIV-1与T细胞区室中的病毒相比，可能对抗逆转录病毒药物具有降低的敏感性;和（e）在单核细胞区室中存在独特的但不是先前描述的HIV-1表型，其支持巨噬细胞而不是T细胞的CCR 5介导的感染。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）病毒组的异质性基因型和表型的发现表明，血液单核细胞中循环的HIV-1代表复制的HIV-1，其可能由受当前ART方案抑制较少的不同组织中的巨噬细胞产生。然而，所有上述基因型和表型结果均来自PCR扩增的HIV-1 DNA，从病人的纯化单核细胞DNA中提取。这些前病毒是否真的复制并产生传染性病毒仍不清楚。该项目将是建立和表征一组新的感染性HIV-1的第一步，该感染性HIV-1将从患者的纯化M/M中分离出来。然后，我们将定义“天然”M/M-HIV-1与M/M谱系细胞（包括CNS相关/衍生的细胞类型）的相互作用。最后，我们将使用这组“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，旨在开发靶向M/M中HIV-1感染的新治疗剂，并预防由HIV-1感染的巨噬细胞引起的CNS损伤。项目叙述：我们将建立和表征一个新的面板的HIV-1从M/M分离，并确定单核细胞衍生的HIV-1与中枢神经系统细胞的相互作用。此外，我们将通过使用这组“天然”M/M衍生的HIV-1，在离体模型中评估目前批准的新药和新药的抗HIV活性，旨在开发新的治疗剂，预防由这些单核细胞衍生的HIV-1感染的巨噬细胞引起的CNS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.