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Blood, Vol. 111, Issue 5, 2744-2754, March 1, 2008
BAFF enhances chemotaxis of primary human B cells: a particular synergy between BAFF and CXCL13 on memory B cells
Blood Badr et al.
111: 2744
Supplemental materials for: Badr et al
Files in this Data Supplement:
- Figure S1. BAFF and CXCL13 do not modify the expression of adhesion molecules on isolated primary B cells (JPG, 136 KB)
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B cells (2 × 106 cells/ml) were cultured for 16h in complete medium with or without 25 ng/ml BAFF and then analyzed for the expression of various adhesion molecule by flow cytometry using CD29-APC (MAR4,  1 integrin), CD44-APC (G44-26), CD49d-APC (9F10,  4 integrin), CD62L-FITC (SK11, L-selectin) or 7-integrin-PE (FIB504) antibodies or their matching isotype controls: FITC-, PE- and APC-conjugated IgG1 (MOPC21), PE-conjugated IgG2a (G155-178) and APC-conjugated IgG2b (27-35) (all from BD Biosciences). LFA-3 (AICD58), LFA-1 (CD18/CD11a, IOT16/IOT18) (from Beckman Coulter, Marseille, France) expression was determined by indirect immunofluorescence using PE-conjugated polyclonal goat F(ab′)2 anti-mouse IgG. Purified mouse IgG1 (11711.11) and IgG2a (20102.1) isotype controls were purchased from R&D systems. Staining with CD20-Cy7PE was also performed and used as a control. (A) Surface expression of CD44, CD20, CD18, 7-integrin, CD29 and CD49d were assessed on mock-treated (gray histogram) and BAFF-treated (open histogram, bold line) B cells. One representative experiment of 4 is shown. (B) Modulation of the expression of each adhesion molecule was calculated as the ratio between the MFI values in BAFF- and in mock-treated cells. Results were obtained using samples from 4 different donors and are expressed as mean values ± SEM. (C) BAFF-treated B cells (for 16 h with 25 ng/mL BAFF) were incubated for 3 hours with medium or 500 ng/mL CXCL13. Surface expression of 7-integrin, CD29 and CD49d were then assessed on mock-treated (gray histogram) and BAFF-treated (open histogram, bold line) B cells. One representative experiment of 2 is shown.
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