TB-associated microenvironment promotes HIV latency in CD4+ T cells

G. DUETTE; A. DE VRIES-EGAN; Z. VAHLAS; M. SHARABAS ; A. PEREYRA CASANOVA; C. MELUCCI; P. PEREYRA GERBER; C. VEROLLET; L. BALBOA; S. PALMER

Miami

Workshop; HIV Persistence Workshop; 2022

BACKGROUND/AIMSHIV-infected individuals are frequently co-infected with Mycobacterium tuberculosis (Mtb)-the causative agent of tuberculosis (TB). The reduction of CD4 T-cells associated with HIV infection increase the risk of active TB. However, the effects of the TB-associated microenvironment on HIV infection and latency in CD4 T-cells is unclear. Therapeutically aspirated pleural effusions from TB patients (TB-PE), can reflect the microenvironment found in human respiratory cavities impacted by Mtb infection. Therefore, we investigated the effects of TB-PE on HIV replication and latency within CD4 T-cells.METHODSCD4 T-cells were isolated from 6 healthy-donors and infected with HIVNL4-3 in the presence or absence of TB-PE. Viral entry was assessed through the HIV-Vpr-BlaM system, reverse transcription was quantified by qPCR, and the percentage of HIV-infection measured by p24 immunostaining. For HIV latency studies, CD4 T-cells from 4 healthy donors were infected with a dual-fluorescent reporter HIV. Cells latently infected with this reporter virus express the near-infrared fluorescent protein (iRFP) while cells undergoing a productive infection express both iRFP and the green fluorescent protein (GFP). The proportion of latently HIV-infected cells, in the presence or absence of TB-PE, was quantified by flow cytometry. To test whether TB-PE affects HIV latency reversal, CD4 J-Lat cells were exposed to PMA with or without TB-PE and the expression of GFP measured by flow cytometry.RESULTSWe observed that the reverse transcription, and percentage of HIV infection in CD4 T-cells decreased in the presence of TB-PE (p< 0.05). However, this was not due to differences in the viral entry. We also observed that the incubation with TB-PE significantly increased the proportion of latently HIV-infected cells (p< 0.05). In addition, PMA-induced latency reversal of CD4 T-cells was significatively reduced with TB-PE treatment (p< 0.05).CONCLUSIONOur results indicate that the presence of TB-PE negatively impacts viral replication, induces HIV latency, and decreases latency reversal in CD4 T-cells. Importantly, our study suggests that TB-associated microenvironments may contribute to HIV persistence and the viral reservoir and pose a challenge to HIV curative strategies, such as shock and kill, which rely on latency reversal in co-infected patients.