A Role for microRNA-155 Modulation in the Anti-HIV-1 Effects of Toll-Like Receptor 3 Stimulation in Macrophages
Figure 5
miR-155 over-expression in primary macrophages and cell lines decreases their susceptibility to HIV-1 infection.
MDMs from 12 different donors were transfected with miR-155 expression plasmid (pEZX-miR-155) or scrambled control (pEZX-scrambled) (GeneCopoeia) using jetPEI-Macrophage transfection reagent (Polyplus), following manufacturer's instructions. After 48 h., cells were either infected with BaL pseudotype (A) to evaluate infection through luciferase activity as indicated above; or total RNA was isolated for miR-155 quantitation by real-time PCR (B) as indicated above. HOS-CD4-CCR5 cells were also transfected with various amounts of the miR-155 or scrambled plasmids using calcium precipitation, and then used for infection with BaL or VSV-G pseudotypes (C) or RNA isolation for miR-155 quantitation (D). Infection was evaluated in 8 replicates as percent luc activity compared to untransfected cells (mean ± SD). Relative miR-155 levels were evaluated as in Figure 3 and results are shown as fold-change (mean ± SD) with respect to the untransfected control.