Figure 4
FSL-1 significantly reduced HSV-2 replication in vitro. Triplicate human vaginal EC cultures were treated with FSL-1 (6 μg or 0.1 μg) at 24 h (black bar), 6 h (grey bar) or just prior (white bar) to HSV-2 inoculation (104pfu/well). A separate set of cultures were treated with the DPBS vehicle, infected and processed in parallel (hatched bar). Following 24 h of viral infection, each well was collected (100 μL) and 50 μL of the sample was used for standard plaque titration assays. DNA was extracted from the remaining sample (50 μL) and subjected to quantitative PCR to measure the amount of viral replication. FSL-1 (6 μg) delivered 24 or 6 h prior to infection significantly (*, p < 0.05; student's t-test) reduced viral replication compared to the DPBS vehicle for both PCR and plaque titration analyses. For the 0.1 μg FSL-1 dose, significant (*, p < 0.05; student's t-test) reduction in GE was observed 24 h prior to HSV-2 inoculation while significant (*, p < 0.05; student's t-test) reduction in pfu was observed 6 h prior to viral challenge. Addition of FSL-1 at either dose just prior to infection did not impact HSV-2 replication. Results are presented as the mean ± SEM from two separate experiments using three vaginal EC lines (V11I, V12I, V19I) in each experiment.