Fig. 2.

Cross-reactivity of memory B cell-derived antibodies from longitudinal samples against 1086.C and 1176.C gp140 Envs. a Flow plots of 1086.C and 1176.C gp140 staining of Env-reactive memory B cells in the indicated animals at 0, 57, and 109 weeks post immunization. Memory B cells which bound both BV421 (x-axis) and AF647 (y-axis) labeled Envs were defined as double positive (DP) Env-reactive memory B cells. Sorting of DP Env-reactive B cells was performed to ensure sort accuracy. No pre-existing cross-reacting memory B cells were present before vaccination (week 0). b Env reactivity of each individual sorted memory B cell was confirmed in ELISA after in vitro expansion and differentiation into antibody-secreting cells as described in the Methods. The Venn diagrams show the percentage of Env-specific supernatants from Ig-secreting cultured memory B cells that bound to 1086.C gp140 (yellow) and/or 1176.C gp140 (blue) Env. The number of Env-specific memory B-cell supernatants screened for each animal at each time point is indicated below each diagram