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Stage-specific oligonucleotide uptake in murine bone marrow B-cell
precursors
Q Zhao, T Waldschmidt, E Fisher, CJ Herrera and AM Krieg
Department of Internal Medicine, University of Iowa, Iowa City.
Fluorescein isothiocyanate (FITC)-conjugated phosphodiester and
phosphorothioate oligonucleotides were used in four-color flow cytometry
with murine bone marrow cells stained with monoclonal antibody specific for
the differentiation markers B220, S7 (CD43), and BP-1 to show possible
stage-specific oligonucleotide uptake. Relatively low uptake was observed
among pre-Pro- and early Pro-B cells. Late Pro- B- and pre-B cells had
increased oligonucleotide uptake, whereas B cells had a lower level. Cell
membrane binding of oligonucleotides varied during B-cell differentiation
in parallel with internalization, which was documented by confocal
microscopy. An FITC-conjugated polyanionic dextran sulfate also showed
differentiation-related B-cell association, suggesting the presence of cell
membrane binding sites specific for polyanions as opposed to a unique
feature of the DNA backbone. Interpretation of antisense experiments in
murine bone marrow cells will need to account for the heterogeneous
oligonucleotide uptake among differentiating B cells.
Volume 84,
Issue 11,
pp. 3660-3666,
12/01/1994
Copyright © 1994 by The American Society of Hematology
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