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. ? ? ?
| Wang, Tong; Xu, Younong; Zhu, Haiying et al. (2013) Successful isolation of infectious and high titer human monocyte-derived HIV-1 from two subjects with discontinued therapy. PLoS One 8:e65071 |
