Figure 2.
Integrin α6 blockade using the anti-α6 mAb P5G10 deadheres primary ALL cells and sensitizes the cells to chemotherapy. Six primary B-ALL cell lines (LAX7R, LAX53, PDX2, TXL3, SFO2, ICN24) and 2 B-ALL cell lines (Kasumi-2 and BEL-1) were preincubated with purified anti-human α6 Ab P5G10 (red bars) or its isotype control IgG1 (white bars) on plates coated with hlaminin-1. Adhesion of ALL cells to the bovine serum albumin (BSA) control, laminin-1 (A-E, left panels), cell culture media (media) or OP9 stroma cells (A-E, right panels) was assessed after overnight incubation. The viability of the ALL cells LAX7R (F), LAX53 (G), PDX2 (H), TXL3 (I), SFO2 (J), and ICN24 (K), plated on BSA (left panel) or laminin-1 (right panel) and treated with or without α6 blockade (P5G10) combined with nilotinib (NTB) or VDL for 2 days is shown. Viability of B-ALL cell lines Kasumi-2 (L) and BEL-1 cells (M) plated on BSA (left panel) or laminin-1 (right panel) treated with VDL combined with P5G10 for 3 days is shown. Three primary ALL cell lines, LAX7R (KRASG12V) (N), SFO2 (BCR-ABL1+) (O), and TXL3 (BCR-ABL1+) (P), were cocultured with OP9 cells and incubated with P5G10 (red bars) or its isotype control IgG1 (white bar) combined with NTB or VDL for 5 days. VDL was used at indicated concentrations (0.005 μM: 5 nM vincristine, 50 pM dexamethasone, and 0.0025 IU L-asparaginase; 0.001 μM: 1 nM vincristine, 10 pM dexamethasone, and 0.005 IU L-asparaginase; 0.05 μM: 50 nM vincristine, 500 pM dexamethasone, and 0.025 IU L-asparaginase). Viability was measured via the exclusion of dead cells based on trypan blue staining. LAX7R cells (Q-R) were plated on laminin-1 or OP9 cells and treated with the following antibodies: α6 function-blocking J8H (gray bars; 20 µg/mL), α6 adhesion- and function-blocking P5G10 (red bars; 20 µg/mL), or IgG1 (white bar; 20 µg/mL). Adhesion (overnight incubation) (Q) and viability (2 days incubation) (R) were determined by trypan blue exclusion staining. Means ± standard deviations are shown. One experiment of 3 is shown. Each experiment was performed in triplicate. *P < .05; **P < .01; ***P < .001 by 1-way analysis of variance (ANOVA) with post hoc analysis (Tukey test). DMSO, dimethyl sulfoxide; NS, not significant.

Integrin α6 blockade using the anti-α6 mAb P5G10 deadheres primary ALL cells and sensitizes the cells to chemotherapy. Six primary B-ALL cell lines (LAX7R, LAX53, PDX2, TXL3, SFO2, ICN24) and 2 B-ALL cell lines (Kasumi-2 and BEL-1) were preincubated with purified anti-human α6 Ab P5G10 (red bars) or its isotype control IgG1 (white bars) on plates coated with hlaminin-1. Adhesion of ALL cells to the bovine serum albumin (BSA) control, laminin-1 (A-E, left panels), cell culture media (media) or OP9 stroma cells (A-E, right panels) was assessed after overnight incubation. The viability of the ALL cells LAX7R (F), LAX53 (G), PDX2 (H), TXL3 (I), SFO2 (J), and ICN24 (K), plated on BSA (left panel) or laminin-1 (right panel) and treated with or without α6 blockade (P5G10) combined with nilotinib (NTB) or VDL for 2 days is shown. Viability of B-ALL cell lines Kasumi-2 (L) and BEL-1 cells (M) plated on BSA (left panel) or laminin-1 (right panel) treated with VDL combined with P5G10 for 3 days is shown. Three primary ALL cell lines, LAX7R (KRASG12V) (N), SFO2 (BCR-ABL1+) (O), and TXL3 (BCR-ABL1+) (P), were cocultured with OP9 cells and incubated with P5G10 (red bars) or its isotype control IgG1 (white bar) combined with NTB or VDL for 5 days. VDL was used at indicated concentrations (0.005 μM: 5 nM vincristine, 50 pM dexamethasone, and 0.0025 IU L-asparaginase; 0.001 μM: 1 nM vincristine, 10 pM dexamethasone, and 0.005 IU L-asparaginase; 0.05 μM: 50 nM vincristine, 500 pM dexamethasone, and 0.025 IU L-asparaginase). Viability was measured via the exclusion of dead cells based on trypan blue staining. LAX7R cells (Q-R) were plated on laminin-1 or OP9 cells and treated with the following antibodies: α6 function-blocking J8H (gray bars; 20 µg/mL), α6 adhesion- and function-blocking P5G10 (red bars; 20 µg/mL), or IgG1 (white bar; 20 µg/mL). Adhesion (overnight incubation) (Q) and viability (2 days incubation) (R) were determined by trypan blue exclusion staining. Means ± standard deviations are shown. One experiment of 3 is shown. Each experiment was performed in triplicate. *P < .05; **P < .01; ***P < .001 by 1-way analysis of variance (ANOVA) with post hoc analysis (Tukey test). DMSO, dimethyl sulfoxide; NS, not significant.

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