Figure 3
Neutralization of three Ad vectors by anti-gp120 (902) MAb or anti-Ad PAb. A) To determine whether the V3-presenting vectors could be neutralized by the anti-gp120 (902) MAb, MAb was diluted into three working concentrations (1:500, 1:1,000 and 1:5,000) and incubated with Ad (negative control), Ad-HVR1-lgs-His6-V3 and Ad-HVR1-long-V3, respectively. Then the mixtures were added onto HeLa cells, followed by incubation for 2 hours. The infectious titer was 5 IP/cell. Samples were harvested at 24 hours post-infection (h.p.i.) and subjected to luciferase signal quantification by Luciferase Assay System Kit (Promega). The data were presented as neutralization ratios. B) To determine whether the V3-presenting vectors and Ad vector could be neutralized by the anti-Ad PAb, PAb was diluted into three working concentrations (1:2,000, 1:4,000 and 1:20,000), and incubated with Ad, Ad-HVR1-lgs-His6-V3 and Ad-HVR1-long-V3, respectively. Samples were harvested at 24 h.p.i. and subjected for measurement of luciferase signal, and the data were presented as neutralization ratios. The values were expressed as the mean ± standard deviation, representing five independent replicates.