A flow cytometry technique to study nuclear factor-kappa B (NFκB) translocation during human B cell activation

We aimed at examining NFkappaB translocation in B lymphocytes during in vitro activation through the specific receptor for antigen using a technique convenient in most laboratories such as flow cytometry. We present here an original, convenient, and reproducible technique to study B cell activation events through NFkappaB translocation by means of a novel, specific flow cytometry assay. Intranuclear translocation of NFkappaB p65 was induced after a 45 min stimulation; the highest signal was detected for a 10 ng/ml stimulus compared to the unstimulated condition (P < 0.05). Purified CD19(+) B cells-cultured in the presence of optimal concentrations of anti-mu fragment Abs (10 ng/ml) for 45 min at 37degreesC-induced a mean 60% (range: 45-67%) MFI- and thus, nuclear NFkappaB translocation-increase. We observed a one-pike profile of NFkappaB staining in PBMC B cells and a two-pike profile of NFkappaB staining in using tonsil B cells. B cells are susceptible to various dysregulations leading to minor to severe pathology (including immunoproliferative disorders). Studies of signal transduction carried out specifically in human B cells, using a novel technique gave considerable advantages: feasibility, sensitivity, reproducibility, ease. (C) 2003 Elsevier B.V. All rights reserved.