FIG 6.

Polyreactivity of vaccine-induced gp41-reactive and gp120-reactive MAbs in DNA/rAd5-vaccinated RMs. (A) Cross-reactivities of representative gp41-reactive (DH785 and DH787) and gp120-reactive (DH798 and DH802) MAbs isolated from blood memory B cells of DNA/rAd5-vaccinated RMs against autologous anaerobic (An) and aerobic (A) fecal bacterial WCLs by SDS-PAGE Western blotting. A total of 50 μg each of anaerobic and aerobic bacterial WCLs was loaded in individual lanes, and MAbs were tested at 20 μg/ml under both nonreducing and reducing conditions. Examples of Western blots of macaque Env MAbs under reducing conditions are shown. (B) Neutralization profile of HIV-1 gp41-reactive and gp120-reactive MAbs. Env-reactive MAbs were screened for HIV-1 neutralization via a TZM-bl assay. The control virus was murine leukemia virus (MuLV). The neutralization color key is shown at the bottom. (C) HEp-2 epithelial cells were stained with gp41-reactive (DH794) and gp120-reactive (DH802) MAbs at a concentration of 50 μg/ml, followed by incubation with FITC-labeled goat anti-human Ab. Images were acquired for 8 or 12 s (DH794 and DH802). + ctl, MAb Ab901062Rh (gp41); − ctl, DH570.30 (gp41). Scale bars, 25 μm. Data are representative of results from at least two separate experiments.