FIG 4.
Neutralizing activities of MAb 2424 versus the other V3 MAbs against JRFL produced in the presence or absence of a mannosidase inhibitor. (A) HIV-1 JRFL pseudoviruses were produced in transfected 293T cells in the absence or presence of 25 μM kifunensine (JRFLKif) and tested for neutralization by V3 MAbs (2424, 447-52D, 2219, 2557, 3869, and 3074), MAb 2G12, CD4bs MAb b12, or an irrelevant MAb, MAb 1418. (B) HIV-1 JRFL pseudoviruses were produced in 293T cells with or without 20 μM swainsonine (JRFLSwain) and tested for neutralization by V3 MAbs 2424 and 447-52D. Serially diluted MAbs were incubated with virus at 37°C for 1 h. TZM.bl cells were then added to the virus-MAb mixtures in complete DMEM containing DEAE-dextran. Neutralization activity was assessed after 48 h. Results with 2424 and 447-52D were averages from duplicate wells, and data from one representative experiment are shown. Results with other MAbs are shown as averages from two to three experiments. P values were determined by the two-way ANOVA. (C) V3 MAb reactivity with gp120 JRFL was assessed in ELISA. Recombinant gp120 protein (1 μg/ml) was coated on 96-well plates and reacted with serially diluted V3 MAbs. Data from a representative experiment with averages and standard deviations from duplicate wells are shown.