Figure 1.

Human β-defensins inhibit HIV-1 replication in macrophages in vitro. MDM were infected with HIV-1_BaL. After virus removal and washing, hBD1, hBD2, and hBD3 (4.7 µM) (A) or increasing concentrations of hBD2 (0-23.3 µM) (B) were added to cultures. Cells were pretreated with AZT as control. Infection was monitored by assaying supernatants for HIV p24 production by ELISA at the times indicated. Data are presented as mean ± SEM of triplicates. Representative experiment, n=3. *P < 0.05, **P < 0.005, ***P < 0.0005, ****P < 0.0001 between treatment and control groups determined with unpaired two-tailed t test. (C) ß-defensins are not toxic to macrophages at the concentrations that inhibit HIV-1. Cells treated with hBDs were tested using MTS assay. Cells were cultured in triplicate in 96-well plates for 3 days in the presence or absence of β-defensins; MTS mix was added and incubated 1 to 4 hrs prior to spectrophotometric absorbance readings at 490 nm. Triplicate readings were averaged (± SEM) and percentage OD ratios of treated/control cells were calculated. (D) hBD2 inhibit infection of MDM with a transmitted-founder HIV strain. MDM were infected with transmitter-founder virus AD17. After virus removal and washing, hBD2 at concentrations indicated above were added to cultures. HIV p24 release in supernatants was monitored by ELISA at the time indicated, and % inhibition was calculated as % of HIV p24 production from untreated MDM. Data are presented as mean ± SEM of triplicates, N=3.