Monitoring HIV-1 treatment in immune-cell subsets with ultrasensitive fluorescence-in-situ hybridisation

Memory CD4 + T Cells Are the Earliest Detectable Human Immunodeficiency Virus Type 1 (HIV-1)-Infected Cells in the Female Genital Mucosal Tissue during HIV-1 Transmission in an Organ Culture System

“…For this purpose, a standard cervical tissue-based organ culture was set up with HIV-1 BAL as described above. At 6, 24, and 96 h after viral exposure tissues were frozen and then analyzed by a modified simultaneous ultrasensitive fluorescence in situ HIV-1 RNA detection and immunophenotyping assay (3,6,22). The photomicrographic results are shown in Fig.…”

“…Our data contrast with the studies of Hu et al (13) in the SIV-rhesus monkey model, in which SIV-infected dendritic cells were detected as the first infected cell types shortly after intravaginal exposure to SIV. However, it is difficult to compare our data with the transmission data from the SIV-macaque model because human cervical biopsies from sexually active women with or without known sexually transmittted diseases generally have greatly elevated numbers of activated T cells compared to non-sexually active women (22) and macaques (B. K. Patterson, unpublished data). These data indicate that the cellular and viral factors responsible for the sexual transmission of HIV-1 could be different than those of SIV.…”

“…Immunophenotyping and ultrasensitive fluorescence in situ hybridization were performed as previously described (3,22 Flow cytometric simultaneous immunophenotyping and ultrasensitive fluorescence in situ hybridization assay on immune cells from disaggregated explant biopsies. A representative portion of cervical explant tissue was disaggregated by using the MediMachine (Becton Dickinson, San Jose, Calif.) as per the manufacturer's instructions.…”

“…Although in chronically HIV-1-infected women T cells, macrophages, and Langerhans cells in cervical tissue are infected with HIV-1 (25,26), analysis of lymph nodes in HIV-infected men during the acute phase of infection indicates that actively virus replication occurs in activated and resting CD4 ϩ T cells (36). A number of studies have demonstrated that productively HIV-1-infected cells in vivo are memory T cells (CD4 ϩ CD45RO ϩ ) that respond to antiretroviral therapy (22,24). There is controversy in the simian immunodeficiency virus (SIV)-rhesus monkey model system about the types of cells that become first infected during sexual transmission.…”

Therapeutic Response of HIV‐1 Subtype C in African Patients Coinfected with eitherMycobacterium tuberculosisor Human Herpesvirus–8

“…By using high-resolution therapeutic monitoring, it is now possible to begin deciphering the complex interplay between HIV-1 and OIs. When combined with new tools that measure viral replication in specific cell types [56][57][58][59] and with mathematical modeling of immune activation and target cell availability [25][26][27][28][29][30][31][32], this approach will provide a powerful tool for unraveling the impact of OIs on HIV-1 disease. An understanding of these processes is likely to be critical to the optimization of treatment strategies in southern Africa, a region where OIs may play a major role not only in the pathogenesis of HIV-1 disease but also in patient response to therapy.…”