Figure 1
Figure 1. Dose- and time-dependent effect of BAFF on B-cell chemotaxis. (A) B cells were incubated for 16 hours with medium (□), 25 ng/mL (▫) or 100 ng/mL (■) BAFF, and 25 ng/mL (▤) or 100 ng/mL () APRIL, and were then tested for migration to 250 ng/mL CCL21, 250 ng/mL CXCL12, or 500 ng/mL CXCL13. Basal migration of B cells was 5% (± 1.4%, medium-treated), 4.7% (± 1.1%, 25 ng/mL BAFF–treated), 5.4% (± 0.5%, 100 ng/mL BAFF–treated), 4.3% (± 2%, 25 ng/mL APRIL–treated), and 5% (± 3%, 100 ng/mL APRIL–treated) Results are expressed as the mean percentage plus or minus SD of cells specifically migrating in response to each chemokine. (B) B cells were incubated for up to 48 hours with medium or 25 ng/mL BAFF. Mock-treated (□) and BAFF-treated (■) B cells were analyzed for migration to 500 ng/mL CXCL13. The experiment was performed using samples from 4 different donors, and basal migration ranged from 3% to 4% and 3.3% to 4.7% for mock- and BAFF-treated B cells, respectively. Results are expressed as the percentage plus or minus SD of specific chemotaxis. *P < .05; **P < .005. (C) B cells were treated for 16 hours with medium (□) or 25 ng/mL BAFF (▫) and analyzed for their migration to 250 ng/mL CCL21, 250 ng/mL CXCL12, and 500 ng/mL CXCL13. Basal migration was 4.4% (± 1.2%) and 4.3% (± 1.1%) for mock- and BAFF-treated B cells, respectively. Results are expressed as the mean percentage plus or minus SD of specifically migrating cells obtained for each donor. *P < .05; **P < .005. (D) B cells were treated for 16 hours with medium (data not shown), 25 ng/mL BAFF (▫), or 100 ng/mL CD40MGL (■) and analyzed for their migration to 250 ng/mL CCL21, 250 ng/mL CXCL12, and 500 ng/mL CXCL13. The experiment was performed using samples from 3 different donors, and basal migration was 3.3% (± 2.1%), 2.3% (± 1.5%), and 1.3% (± 0.6%) for mock-, BAFF-, and CD40MGL-treated B cells, respectively. Results are expressed as mean fold increase plus or minus SD. (E) The percentage of B-cell recovery after overnight culture with medium or 25 ng/mL BAFF was calculated for each donor. Results are expressed as median values obtained from 28 independent experiments. Error bars correspond to 95% confidence intervals about the median. (F) B cells from 3 different donors were incubated for 16 hours at 37°C with medium (□) or 25 ng/mL BAFF (▫) and then incubated for 60 minutes at 37°C with medium or 100 ng/mL chemokines. Cells were washed in ice-cold medium and stained with PE-conjugated CCR7, CXCR4, or CXCR5 mAb for 30 minutes at 4°C. The mean channel fluorescence intensity (MFI) values for mock-treated cells were between 57 and 123 for CCR7, 356 and 1067 for CXCR4, and 64 and 81 for CXCR5. Each of these individual values was considered as 100% expression. Data are expressed as the mean plus or minus SD percentage of MFI values for remaining surface expression.

Dose- and time-dependent effect of BAFF on B-cell chemotaxis. (A) B cells were incubated for 16 hours with medium (□), 25 ng/mL (▫) or 100 ng/mL (■) BAFF, and 25 ng/mL (▤) or 100 ng/mL () APRIL, and were then tested for migration to 250 ng/mL CCL21, 250 ng/mL CXCL12, or 500 ng/mL CXCL13. Basal migration of B cells was 5% (± 1.4%, medium-treated), 4.7% (± 1.1%, 25 ng/mL BAFF–treated), 5.4% (± 0.5%, 100 ng/mL BAFF–treated), 4.3% (± 2%, 25 ng/mL APRIL–treated), and 5% (± 3%, 100 ng/mL APRIL–treated) Results are expressed as the mean percentage plus or minus SD of cells specifically migrating in response to each chemokine. (B) B cells were incubated for up to 48 hours with medium or 25 ng/mL BAFF. Mock-treated (□) and BAFF-treated (■) B cells were analyzed for migration to 500 ng/mL CXCL13. The experiment was performed using samples from 4 different donors, and basal migration ranged from 3% to 4% and 3.3% to 4.7% for mock- and BAFF-treated B cells, respectively. Results are expressed as the percentage plus or minus SD of specific chemotaxis. *P < .05; **P < .005. (C) B cells were treated for 16 hours with medium (□) or 25 ng/mL BAFF (▫) and analyzed for their migration to 250 ng/mL CCL21, 250 ng/mL CXCL12, and 500 ng/mL CXCL13. Basal migration was 4.4% (± 1.2%) and 4.3% (± 1.1%) for mock- and BAFF-treated B cells, respectively. Results are expressed as the mean percentage plus or minus SD of specifically migrating cells obtained for each donor. *P < .05; **P < .005. (D) B cells were treated for 16 hours with medium (data not shown), 25 ng/mL BAFF (▫), or 100 ng/mL CD40MGL (■) and analyzed for their migration to 250 ng/mL CCL21, 250 ng/mL CXCL12, and 500 ng/mL CXCL13. The experiment was performed using samples from 3 different donors, and basal migration was 3.3% (± 2.1%), 2.3% (± 1.5%), and 1.3% (± 0.6%) for mock-, BAFF-, and CD40MGL-treated B cells, respectively. Results are expressed as mean fold increase plus or minus SD. (E) The percentage of B-cell recovery after overnight culture with medium or 25 ng/mL BAFF was calculated for each donor. Results are expressed as median values obtained from 28 independent experiments. Error bars correspond to 95% confidence intervals about the median. (F) B cells from 3 different donors were incubated for 16 hours at 37°C with medium (□) or 25 ng/mL BAFF (▫) and then incubated for 60 minutes at 37°C with medium or 100 ng/mL chemokines. Cells were washed in ice-cold medium and stained with PE-conjugated CCR7, CXCR4, or CXCR5 mAb for 30 minutes at 4°C. The mean channel fluorescence intensity (MFI) values for mock-treated cells were between 57 and 123 for CCR7, 356 and 1067 for CXCR4, and 64 and 81 for CXCR5. Each of these individual values was considered as 100% expression. Data are expressed as the mean plus or minus SD percentage of MFI values for remaining surface expression.

Close Modal
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal