The HIV-induced mitochondrial ROS and cell death in hepatic stellate cells contribute to a profibrotic profile

CEVALLOS CINTIA; JARMOLUK PATRICIO; SVIERCZ FRANCO; LOPEZ ALICIA; FREIBERGER NICOLE; GUANO ALEX; DELPINO M. VICTORIA; QUARLERI JORGE

Congreso; 29. The 18th international symposium n viral hepatitis and liver diseases.; 2023

Background: Hepatic stellate cells (HSC) express functional CD4 and CXCR4 receptors on their surfaces, making them susceptible to HIV infection. Whether HIV is able to activate HSCs by damage-related stimuli to secrete excessive extracellular matrix, leading to collagen deposition is undefined. Besides, HSCs may suffer HIV-induced programmed cell death (PCD), which might cause the microenvironment to be shifted toward cellular activation. Since the level of reactive oxygen species (ROS) increases sharply in activated HSCs and HIV exposure may trigger ROS early after viral exposure, our goal is to define the HIV role in propitiating a profibrotic profile by inducing ROS imbalance and PCD after the HSC challenge.Methods: Both cell-free virus (pNL43-GFP) and HIV-infected T lymphocytes (HIV-GFP+ Jurkat cells labeled with violet proliferation dye) were used to examine the LX-2´s permissiveness to HIV replication. Additionally, LX-2 were subjected to conditioned media (supernatant from HIV-infected T lymphocytes). Viral replication and the effectiveness of HIV infection were measured 3 days after exposure (dpe) using the HIV-p24 antigen by ELISA in supernatants and GFP+ cells by flow cytometry, respectively. In LX-2, the PCD (Annexin-V/7AAD) and ROS levels were measured using flow cytometry. Collagen deposition in the extracellular matrix was determined by Sirius Red staining and spectrophotometric quantification. All experiments were carried out in triplicate.Results: At 3 dpe, neither the cell-free virus nor the cell-to-cell contact challenge produced HIV progeny. Cell-to-cell contact with HIV-infected T-lymphocytes significantly enhanced late apoptosis/necrosis in LX-2 compared to control (1.41% vs. 2.0%; p 0.01), albeit not with the cell-free virus. This was also accompanied by an increase in mitochondrial ROS generation (1.29% vs. 2.26%, p 0.01). Additionally, the extracellular matrix of these cells has seen a 1.6x change in collagen deposition (p = 0.01). When LX-2 were exposed to conditioned media, these anomalies were not noticed (from HIV-infected T lymphocytes).Conclusion: Although HSCs are susceptible but not permissive to HIV infection, their cell-to-cell contact with HIV-infected T lymphocytes results in oxidant stress and enhanced cell death that favors an exacerbated deposition of extracellular matrix, ultimately resulting in a profibrotic profile.