CELLULAR CROSS-TALK AND MODULATION OF THE HEPATIC PROFIBROTIC PROFILE IN THE CONTEXT OF HIV-HCV COINFECTION

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

Paris

Simposio; The 18th international symposium n viral hepatitis and liver diseases; 2023

GHS

Abstract Content: Background: Coinfection with the human immunodeficiency virus (HIV) in patients with chronic hepatitis C virus (HCV) infection significantly accelerates liver damage, favoring the progression of fibrosis. After the injury, mediators secreted by damaged hepatocytes and from the microenvironment (eg, T lymphocytes -LT-), maintain an environment of chronic inflammation that promotes activation of hepatic stellate cells (HSC) capable of producing extracellular matrix and promoting liver fibrosis. The mechanism of the acceleration in the progression of liver fibrosis dueto coinfection has not been defined.Purpose: To evaluate the ability of HIV and HCV to modulate programmed cell death (PCD) and mitochondrial homeostasis of HSCs, giving them a profibrotic profile.Methods: Using a fluorescent HIV (pNL43-GFP), the permissiveness of HSCs (LX2) to infection was assessed after challenging them for 5 hours with: (i) free virus (HIV-GFP+), and (ii) cell-cell infection (co-culture with LT-HIV+). After 72h in culture, the kinetics of HIV replication was evaluated by quantifying the HIV-p24 antigen in supernatants, and the infection efficiency by quantifying GFP+ cells by flow cytometry. The effect of HCV was assessed by exposing the LX2/LTHIV co-cultures to a conditioned medium (hepatocyte-HCV+ supernatant or “SN-HCV”) immediately after co-cultivation.Using flow cytometry on LX2, the level of PCD (Annexin-V/7AAD) and the generation of mitochondrial reactive oxygen species –mROS- were defined. Collagen production was determined by Sirius Red staining and subsequent spectrophotometric reading.Results: LX2 were not susceptible or permissive to HIV infection by cell-free virus, and cell-cell contact, resulting in undetectable production of p24 or GFP expression in them. Exposure to HIV of LX2 did not affect PCD compared to control (6.1 vs. 8.6%, p>.05) nor did it affect after exposing them to SN-HCV (6.1 vs. 8.1%, p>.05). However, PCD in LX2 was significantly increased after simultaneous exposure to LT-HIV+ and SN-HCV (11.33%, p