Stage-specific oligonucleotide uptake in murine bone marrow B-cell precursors.
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.
Zhao
Q
Q
Department of Internal Medicine, University of Iowa, Iowa City.
Waldschmidt
T
T
Fisher
E
E
Herrera
C J
CJ
Krieg
A M
AM
D000818:Animals; D001402:B-Lymphocytes / Q000378:metabolism*; D001483:Base Sequence; D001853:Bone Marrow / Q000378:metabolism*; D001854:Bone Marrow Cells; D002454:Cell Differentiation; D002478:Cells, Cultured; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate; D006412:Hematopoietic Stem Cells / Q000378:metabolism*; D051379:Mice; D008811:Mice, Inbred DBA; D008814:Mice, Inbred NZB; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000378:metabolism*; D013696:Temperature
D009841:Oligonucleotides; D016650:Fluorescein-5-isothiocyanate