Figure 2
Figure 2. CD37-SMIP–induced cell death in B-CLL cells. (A) CD37-SMIP–induced cell death in CLL B cells. CLL B cells were treated with media, goat anti–human IgG (α-Fc), trastuzumab with α-Fc, CD37-SMIP with α-Fc, rituximab with α-Fc, or alemtuzumab with α-Fc for 24 hours. The cells were stained with FITC-annexin V and propidium iodide. The numbers shown in each panel represent percentage Annexin V+ and/or PI+ cells. Shown is a representative result from 14 patient samples analyzed. (B) Dose- and time-dependent induction of cytotoxicity by CD37-SMIP. B-CLL cells were treated with indicated concentrations of CD37-SMIP with cross-linker α-FC. The direct cell death at indicated time points was assessed by FITC-Annexin V/PI staining. Percentage cytotoxicity shown represents Annexin V+ and/or PI+ cells normalized with the media control. Error bars represent standard deviation (SD) among 4 B-CLL patient cell samples. (C) Summary of CD37-SMIP–induced direct cytotoxicity. B-CLL cells isolated from 14 patients were subjected to indicated treatment for 24 hours with or without cross-linker (α-Fc). Cell death was examined with FITC-Annexin V and PI. Percentage cytotoxicity shown represents Annexin V+ and/or PI+ cells normalized with the media control. Error bars represent SD among 14 B-CLL patient samples. Statistical analysis of difference between antibody treatments is shown by P values: *, P < .001; **, P < .001; **, P = .04. (D) Correlation of CD37 expression level and direct cytotoxicity by CD37-SMIP. MFI (CD37): mean fluorescence intensity of CD37 staining of different B-CLL cell samples. Percentage cytotoxicity shown represents Annexin V+ and/or PI+ cells normalized with the media control. (E) Flow cytometric analysis showing selective binding of CD37-SMIP to CD19+ but not CD3+ human PBMC. (F) CD37-SMIP–induced cytotoxicity in B but not T cells. Results shown are summary of FITC-Annexin V+ and/or PI+ cells normalized with media control in normal T cells (open bar; n = 5) or B-CLL cells (filled bar; n = 15) 24 hours after treatment with CD37-SMIP or trastuzumab. Error bars represent SD among samples.

CD37-SMIP–induced cell death in B-CLL cells. (A) CD37-SMIP–induced cell death in CLL B cells. CLL B cells were treated with media, goat anti–human IgG (α-Fc), trastuzumab with α-Fc, CD37-SMIP with α-Fc, rituximab with α-Fc, or alemtuzumab with α-Fc for 24 hours. The cells were stained with FITC-annexin V and propidium iodide. The numbers shown in each panel represent percentage Annexin V+ and/or PI+ cells. Shown is a representative result from 14 patient samples analyzed. (B) Dose- and time-dependent induction of cytotoxicity by CD37-SMIP. B-CLL cells were treated with indicated concentrations of CD37-SMIP with cross-linker α-FC. The direct cell death at indicated time points was assessed by FITC-Annexin V/PI staining. Percentage cytotoxicity shown represents Annexin V+ and/or PI+ cells normalized with the media control. Error bars represent standard deviation (SD) among 4 B-CLL patient cell samples. (C) Summary of CD37-SMIP–induced direct cytotoxicity. B-CLL cells isolated from 14 patients were subjected to indicated treatment for 24 hours with or without cross-linker (α-Fc). Cell death was examined with FITC-Annexin V and PI. Percentage cytotoxicity shown represents Annexin V+ and/or PI+ cells normalized with the media control. Error bars represent SD among 14 B-CLL patient samples. Statistical analysis of difference between antibody treatments is shown by P values: *, P < .001; **, P < .001; **, P = .04. (D) Correlation of CD37 expression level and direct cytotoxicity by CD37-SMIP. MFI (CD37): mean fluorescence intensity of CD37 staining of different B-CLL cell samples. Percentage cytotoxicity shown represents Annexin V+ and/or PI+ cells normalized with the media control. (E) Flow cytometric analysis showing selective binding of CD37-SMIP to CD19+ but not CD3+ human PBMC. (F) CD37-SMIP–induced cytotoxicity in B but not T cells. Results shown are summary of FITC-Annexin V+ and/or PI+ cells normalized with media control in normal T cells (open bar; n = 5) or B-CLL cells (filled bar; n = 15) 24 hours after treatment with CD37-SMIP or trastuzumab. Error bars represent SD among samples.

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