Soluble factors from R5- or X4 tropic HIV-1 infected PBMCs activate human hepatic stellate cells.

<p>LX2 cells were treated with virus-free culture supernatants from HIV-1 infected (R5- or X4- tropic) human PBMCs for 72 hr. Mock (M) represents control group that was treated with supernatants from uninfected PBMCs. (A) Total RNA was isolated from LX2 cells and used for the expression analysis of various pro-fibrogenic and pro-inflammatory marker genes using qRT-PCR, as described in Methods. Data represents mean ± S.E. of three independent experiments. *p<0.05 and **p<0.01 vs mock. (B) LX2 cells were treated with culture supernatants from HIV-1 infected PBMCs for 24 hr as described in Methods. Total cell lysates were analyzed for TIMP-1 expression by western blotting. GAPDH was used as a loading control. Data representative of one of the three experiments is shown. (C) LX2 cells were plated in a 96-well plate at passage 4 and serum starved for 24 hr. The cells were then incubated for 72 hr with culture supernatants from X4-, R5- or mock- infected PBMCs. Following treatment, cells were assayed for proliferation and cell survival using the MTT assay. Data represents mean ± S.E. of three independent experiments.