Figure 1

In vitro biological activity of monoclonal anti-CD5, anti-CD71 and anti-HLA-DR antibodies. A, Induction of direct cell death or apoptosis measured after 24 hours treatment of JOK1-5.3 cells with 10 μg/ml mAbs, using the vital dye alamar blue. Results are % dead cells relative to control. CD5 = anti-CD5 mAb; DR = anti-HLA-DR mAb and CD71 = anti-CD71 mAb throughout. B, Effect on long term growth of JOK1-5.3, measured after 3 or 6 days treatment with 10 μg/ml antibodies and alamar blue. Data are % of live cells relative to untreated cells (CTRL) and are representative of at least 3 experiments. C, Percent proliferation of JOK1-5.3 cells, after incubation with single mAbs anti-CD5 or anti CD71 or the two mAbs combination, at a identical total concentration of 75 μg/ml, for 24 or 72 hours, as compared to untreated control cells, determined by incorporation of the radio-labelled thymidine analogue, ¹²⁵I-Iododeoxyuridine. D, Induction of ADCC of JOK1-5.3 cells in presence of 0.01, 0.1 or 1 μg/ml antibodies as indicated, using purified NK cells at a 10:1 effector to target ratio. E, Comparison of ADCC of JOK1-5.3 cells in presence of single mAbs or pairs of anti-CD5+anti-HLA-DR, anti CD5+anti-CD71, or anti-CD71+anti-HLA-DR, at the total concentrations of 2, 10 or 30 ng/ml, using NK cells at a 10:1 effector to target ratio. F, Induction of phagocytosis of B-CLL targets by in vitro differentiated macrophages with 0.1 μg/ml antibodies. All data are representative of at least 3 experiments. G, Complement dependent cell lysis of JOK1-5.3 cells with 10 μg/ml antibodies and 20% human serum.