CD6 Ligation Modulates the Bcl-2/Bax Ratio and Protects Chronic Lymphocytic Leukemia B Cells From Apoptosis Induced by Anti-IgM by Lyda M. Osorio, Angelina.

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CD6 Ligation Modulates the Bcl-2/Bax Ratio and Protects Chronic Lymphocytic Leukemia B Cells From Apoptosis Induced by Anti-IgM by Lyda M. Osorio, Angelina De Santiago, Miguel Aguilar-SantelisesHå, kan Mellstedt, and Mikael Jondal Blood Volume 89(8):2833-2841 April 15, 1997 ©1997 by American Society of Hematology

Upregulation of CD6 expression on B-CLL cells by activation with SAC plus IL-2 and TPA. B-CLL cells from L-156, L-38, and L-194 patients were cultured with or without SAC (0.005%) plus IL-2 (100 U/mL) or TPA (50 nmol/L) for 72 hours and analyzed for CD6 exp... Upregulation of CD6 expression on B-CLL cells by activation with SAC plus IL-2 and TPA. B-CLL cells from L-156, L-38, and L-194 patients were cultured with or without SAC (0.005%) plus IL-2 (100 U/mL) or TPA (50 nmol/L) for 72 hours and analyzed for CD6 expression with IOR-T1 MoAb by flow cytometry. The dashed line represents background fluorescence and the solid line represents CD6 fluorescence. CD6-specific MFI is in the upper right corner of each panel. The x-axis corresponds to logarithmic fluorescence intensity and the y-axis to cell number. Lyda M. Osorio et al. Blood 1997;89:2833-2841 ©1997 by American Society of Hematology

Lyda M. Osorio et al. Blood 1997;89:2833-2841 ©1997 by American Society of Hematology

Lyda M. Osorio et al. Blood 1997;89:2833-2841 ©1997 by American Society of Hematology

Anti-CD6 can prevent B-CLL cells from anti-IgM–induced DNA fragmentation. Anti-CD6 can prevent B-CLL cells from anti-IgM–induced DNA fragmentation. B-CLL cells from patient L-204 were incubated in RAM-Ig–coated 24-well plates with medium, anti-IgM, and/or IOR-T1. DNA fragmentation was determined after 16 hours using PI staining and FACS analysis. The percentage of apoptotic nuclei corresponding to M1 marker is in the left corner of each panel. Lyda M. Osorio et al. Blood 1997;89:2833-2841 ©1997 by American Society of Hematology

Anti-IgM– and IOR-T1-induced effects are not mediated by Fc receptor. Anti-IgM– and IOR-T1-induced effects are not mediated by Fc receptor. B-CLL cells 2 × 106/mL were incubated for 16 hours in RAM-Ig–coated 24-well plates with medium, anti-IgM, IOR-T1, mouse IgG2a, or mouse IgG1 MoAbs in combination as indicated. The percentage of apoptotic nuclei was determined using Hoechst staining. Lyda M. Osorio et al. Blood 1997;89:2833-2841 ©1997 by American Society of Hematology

(A) Expression of Bcl-2 and Bax protein in B-CLL cells from L-206, L-194, and L-156 patients. (A) Expression of Bcl-2 and Bax protein in B-CLL cells from L-206, L-194, and L-156 patients. Cells were incubated in RAM-Ig–coated 24-well plates with medium, anti-IgM, or anti-IgM plus IOR-T1. Bcl-2 and Bax proteins were determined by flow cytometry after 16 hours. The solid line indicates Bcl-2 or Bax staining and the dotted line represents background control cells staining with nonspecific hamster IgG or rabbit IgG. Specific MFI (after the subtraction of background MFI) is in the right corner of each panel. (B) Negative control for Bcl-2 staining. The dotted and solid lines indicate background and Bcl-2 staining in Daudi cell line, respectively. (C) Specificity of the Bax antibody. Two different B-CLL cells were stained with anti-Bax antibodies that had been blocked with the specific peptide (AA 43-61; dashed line). The dotted and solid lines indicate background control and Bax staining, respectively. The x-axis corresponds to logarithmic fluorescence intensity and the y-axis corresponds to cell number. Lyda M. Osorio et al. Blood 1997;89:2833-2841 ©1997 by American Society of Hematology