Figure 6

Effect of individual IGHV4-34 reversions on the binding of subset 4 IG 240 to viable human B cells. (A) Effect of light chain reversion on binding to tonsil B lymphocytes. Tonsil mononuclear cells were incubated with biotinylated versions of CLL subset 4 mAb240 and the original 240 H chain paired with the L chain revertant rL-1, followed by incubation with streptavidin PE-Cy7. Note the significant drop in memory B cell binding and the increase in naïve B cell binding. (B) Effect of light chain reversion or light chain swapping on binding to Ramos B cells. Ramos B cell line cells were incubated with unmanipulated CLL subset 4 mAb240 or the original 240 H chain paired with the L chain revertant rL-1 (left panel) or with unmanipulated CLL subset 4 mAb183 or the original 183 H chain paired with the L chain rearrangement from CLL mAb 141, an IGHV4-34⁺ IG that is not a member of subset 4 (right panel). In both cases (left and right panels), each mAb was followed by incubation with streptavidin PE-Cy7. See Figure 5 for explanation of abbreviation for rL-1 revertant.