Figure 4

Reactivity of anti-macaque CD38 mAbs with native and DTT-treated NIH/macCD38 and NIH/humCD38. A. Flow cytometric profile of NIH/mac38 cells treated for 45 min at 37°C with 10 mM DTT (red profile) or without DTT (black profile) and then tested for binding of anti-cynomolgus CD38 mAbs by IF. Grey profile shows reactivity of CBT3G mAb (isotype control). B. Results of IF/DTT experiments illustrated by confocal microscopy. Transfectants are indicated at the top of the panel. Left vertical triplet of images (from top to bottom): (1) reactivity of mAb KK1B5 with native cynomolgus CD38 cells stained with FITC; (2) reactivity with DTT-treated cells stained by FITC; (3) cells in plate 2 viewed by differential interference contrast (DIC). (4–6): idem mAb KK1B5 with NIH/humCD38; (7–9): idem mAb KK4E5 mAb with NIH/macCD38. C. IF/confocal microscopy of NIH/humCD38, with/without DTT. Left vertical triplet of images (from top to bottom) shows staining with mAb IB4 and FITC of control (1), DTT-treated visualized by FITC (2) and DIC (3). Right trio: results with mAb AT1 (4–6). D. Flow cytometric profiles of control (black profile) and DTT-treated (red profile) NIH/macCD38 binding by anti-human CD38 mAbs and FITC. Grey profile shows reactivity of CBT3G mAb (isotype control).