Bystander Effects and Profibrotic Interactions in Hepatic Stellate Cells during HIV and HCV Coinfection

Figure 1

Experimental timeline schedule. The permissiveness of HSCs (LX-2) to HIV infection was assessed using two strategies: (I) cell-free virus (pNL43-GFP) and (IIa) HIV-infected Jurkat cells with pNL43-GFP in coculture with LX-2 (ratio 5:1). LX-2 cells were exposed to HIV during 5 hr and washed. After 72 hr in culture, the kinetics of HIV replication was evaluated on LX-2 cells by quantifying the HIV-p24 antigen in supernatants utilizing an ELISA kit, and the infection efficiency by quantifying GFP+ cells by flow cytometry was evaluated. (IIb) Moreover, the effect of supernatant collected at 72 hr from HIV-infected LT CD4+ cells (Jurkat)—“cm Jurkat (HIV)”—on LX-2 was evaluated. Complementary, (III) the effect of HCV was assessed by exposing the LX-2/LT HIV cocultures to a conditioned medium (supernatant collected at 72 hr from HCV-infected hepatocytes (Huh7.5)—“cm Huh/HCV”—immediately after cocultivation. The different determinations on LX-2 cells were performed at 2 or 72 hr post-coculture or exposure to conditioned medium. The percentage of infection on Jurkat and Huh7.5 cells was evaluated at 72 hr postinfection and immediately before cocultivation or conditioned medium stimulation. LX-2 and Jurkat cells were infected to cell-free virus (1 pg/cell) and Huh7.5 cells were infected with a MOI: 1.