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

Figure 4

Determination of profibrotic and proinflammatory parameters. (a) Determination of IL-6 secretion by LX-2 cells implementing ELISA kit. Seventy-two hours post-cocultivation with Jurkat (HIV), the IL-6 production was higher than control (LX-2 cocultivated with uninfected Jurkat) . (b, c) Determination of α-SMA production by LX-2 revealed by immunofluorescence with a specific antibody and real-time PCR. The α-SMA production (protein expression and mRNA levels) was increased 1.2-fold when LX-2 cells were cocultured with Jurkat (HIV) exposure to “cm” from Jurkat (HIV) in comparison with the corresponding controls (LX-2 cocultivated with uninfected Jurkat and LX-2 exposure “cm” from uninfected Jurkat). (d) In the same way, the intracellular production of TGF-β by LX-2 cells (determined by flow cytometry) was increased up to 4.3-fold for both conditions from HIV-infected Jurkat cells (“cm” and cell-to-cell contact). (e–h) Determination of collagen deposition by LX-2 cocultured with Jurkat (HIV) or exposure to conditioned medium “cm” from Jurkat (HIV) was a threefold increase compared to corresponding controls. Such determination was performed utilizing immunofluorescence microscopy and Sirius Red staining. (i) MMP-9 levels quantified in LX-2 cocultured with Jurkat (HIV) or exposure to “cm” from Jurkat (HIV) culture supernatant implementing an ELISA kit were downregulated (4.3-fold). NI, noninfected; cm, conditioned medium); and NS, not significant). Scale bar: 50 µm. Graphics are showing values obtained from three independent experiments. Data are given as the mean ± SD. , , and .

(a)

(b)

(c)

(d)

(e)

(f)

(g)

(h)

(i)