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Expression and visualization during cell cycle progression of alkaline phosphatase in B lymphocytes from C3H/HeJ mice
Author(s) -
Souvannavong Vongthip,
Brown Spencer,
Sarih M'hammed,
Adam Arlette
Publication year - 1994
Publication title -
journal of leukocyte biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.819
H-Index - 191
eISSN - 1938-3673
pISSN - 0741-5400
DOI - 10.1002/jlb.55.5.626
Subject(s) - biology , alkaline phosphatase , flow cytometry , microbiology and biotechnology , cell , cytoplasm , cell sorting , cell cycle , enzyme , lipopolysaccharide , acid phosphatase , biochemistry , immunology
The expression of alkaline phosphatase (APase) activity by purified B cells from lipopolysaccharide (LPS)‐hyporesponsive C3H/HeJ mice was determined. Optimal APase activity was expressed after costimulation with interleukin‐5 and dextran sulfate (DXS), whereas LPS, which is highly effective on B lymphocytes from normal mice, was unable to induce enzyme expression, even in the presence of DXS. The simultaneous determination by flow cytometry of both cellular APase, by using a fluorescent azo dye technique, and DNA content showed that APase was highly expressed by about one‐tenth of cells in G1 phase, whereas it was present in more than 50% of cells in S and G2/M phases. The enzyme, as visualized by confocal microscopy after cell sorting on the basis of DNA content, was found to be, localized mainly in vesicular structures distributed throughout the cytoplasm in G1 cells. It was distributed in patches and essentially localized at the cell periphery in S cells, whereas clear capping of activity was observed in G2/M cells. J. Leukoc. Biol. 55: 626–632; 1994.