Figure 5

A functional ADAM10 metalloprotease domain is not required for HIV-1 replication. (A) Over-expression of wt ADAM10 in U373-MAGI-CCR5 cells increased HIV-1 replication; however, cell toxicity was noted at concentrations of 0.5 μg and above. β-galactosidase activity was measured 48 h after infection (*P < 0.05, **P < 0.01). (B) Over-expression of wt ADAM10 and ADAM10 E384A both increased HIV-1 replication U373-MAGI-CCR5 cells. 0.5 μg of DNA plasmid was transfected into U373 cells and infected with HIV-SF162 48 hours after transfection. Cells were lysed and β-galactosidase activity was measured 48 h after infection (*P < 0.05). (C) Serial dilutions of tissue inhibitors of metalloprotease 1 (TIMP-1) were added to primary macrophages, and cell viability was assessed 24 h after addition of TIMP-1. (D) TIMPs had no effect on HIV-1 replication in primary human macrophages. TIMPs (25 nM) were added to primary macrophages 24 h prior to and during infection with HIV-SF162. Supernatant was collected 7 d after infection and HIV-1 p24 production was measured by ELISA.