SARS‐CoV‐2‐Induced Macrophage Polarization Reverses HIV‐1 Latency in J‐Lat Cells Through TNFα Signaling

Abstract

Introduction: Coronavirus disease 2019 (COVID-19) may have both short- and long-term impacts on the progression of human immunodeficiency virus (HIV)-1 after acute SARS-CoV-2 infection in people living with HIV (PLWH), even those on combined antiretroviral therapy (cART). This study aimed to investigate whether SARS-CoV-2 could influence HIV reactivation in latently infected lymphoid cells.Methods: HIV-infected lymphoid (J-Lat) cells, characterized by proviral latency under unstimulated conditions, were used for latency reversal assays with phorbol 12-myristate 13-acetate (PMA), free SARS-CoV-2 particles, or conditioned media (CM) from macrophages. Monocytes isolated from donor blood were differentiated into macrophages (monocyte-derived macrophages [MDMs]) and polarized to M1 or M2 phenotypes before stimulation or with two SARS-CoV-2 variants (wild-type and BA.5) infection. Additionally, the effects of redox imbalance on latency reversal in both J-Lat and myeloid (U1) latency models were measured. SARS-CoV-2 RNA was quantified by RT-qPCR targeting ORF1ab and N genes, and latency reversal and reactive oxygen species (ROS) levels were assessed by flow cytometry. TNFα involvement was confirmed through neutralization assays, while cytokines and polarization markers were analyzed via ELISA and fluorescent antibodies.Results: Jurkat and J-Lat cells had low ACE2 expression and were not permissive to SARS-CoV-2 infection. SARS-CoV-2 exposure alone did not induce HIV latency reversal in J-Lat cells. However, CM from M1-polarized, resiquimod (R-848)-treated, and SARS-CoV-2-infected macrophages significantly reactivated latent HIV. TNFα was identified as the primary driver of latency reversal, with no significant changes in ROS levels. Prolonged SARS-CoV-2 exposure shifted macrophage polarization toward an anti-inflammatory M2 phenotype, characterized by IL-10 release, which reduced latency reactivation.Conclusions: This study demonstrates that SARS-CoV-2 can indirectly reverse HIV latency in lymphoid cells by promoting the release of pro-inflammatory cytokines from infected macrophages. These findings suggest potential therapeutic strategies for preventing HIV reactivation during SARS-CoV-2 coinfection, emphasizing the modulation of cytokine signaling to control inflammation while minimizing immune dysregulation.