A simplified method for Ca2+ flux measurement on isolated human B cells that uses flow cytometry.

Clinical and Diagnostic Laboratory Immunology
L GergelyV Agnello

Abstract

A method for Ca2+ flux measurement on isolated human peripheral B cells that uses flow cytometry is described. B cells were isolated by anti-CD19 magnetic bead sorting, and Ca2+ flux was measured with the fluo-3 reagent on a standard single-laser flow cytometer. The response of B-cell stimulation by anti-immunoglobulin B (anti-IgM), anti-IgD, protein A, concanavalin A, and ionomycin was determined. Percentage of responder B cells, the level of Ca2+, and the time of peak stimulation were measured. Bound anti-CD19 monoclonal antibody coupled with small paramagnetic particles did not affect Ca2+ flux. All the isolated B cells responded maximally at 10s with stimulation by 8 microg of ionomycin. The average isolated preparation contains 70% IgM+ and 85% IgD+ cells, all of which showed peak stimulation with 10 microg of anti-IgM and anti-IgD per ml, respectively, at 30s. Only at high concentrations of 80 microg/ml, concanavalin A produced a slower response, peaking at 90 s after stimulation. Stimulation with 20 microg of protein A per ml resulted in Ca2+ flux in only 40 to 60% of cells that had a rapid response and maximal stimulation resembling the pattern of activation of ionomycin. B cells from three patients with mixed cryoglobu...Continue Reading

References

Nov 19, 1992·The New England Journal of Medicine·V AgnelloL M Kaplan
Jan 1, 1990·Cytometry·S MiltenyiA Radbruch
Dec 1, 1990·Japanese Journal of Pharmacology·T Yamauchi, I Yamamoto
Jan 1, 1990·Cytometry·G T RijkersJ C Cambier
Mar 20, 1993·Annals of the New York Academy of Sciences·P S RabinovitchT J Kavanagh

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Citations

Sep 13, 2007·Analytical and Bioanalytical Chemistry·Xunli ZhangStephen J Haswell
Apr 10, 2021·Cell Communication and Signaling : CCS·Helena M StrickerThomas Dittmar
Dec 25, 2002·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Katrien PrincenDominique Schols
Aug 14, 2019·JCI Insight·Soizic GaraudKaren Willard-Gallo
Jan 26, 2006·American Journal of Reproductive Immunology : AJRI·Noemi KozmaJulia Szekeres-Bartho

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