Uptake of oligodeoxyribonucleotides by lymphoid cells is heterogeneous and inducible.
Oligonucleotide uptake was studied in cultured murine spleen and lymph node cells using internally radiolabeled and fluorescein-5-isothiocyanate (FITC)-labeled oligonucleotides. Lymphoid subpopulations were distinguished by flow cytometry and staining with antibodies to cell-surface molecules. Approximately 5% of fresh lymphoid cells take up substantial amounts of oligonucleotide. The percentage of B cells that take up oligonucleotide increased fivefold if cells were cultured for at least 24 hr prior to incubation with labeled oligonucleotides, and increased 10-fold if cells were precultured for 48 hr. T-cell uptake changed very little in culture. Cultured CD4+ and CD8+ T cells had similar oligonucleotide uptake that was less than one-third of that in cultured B cells, but CD4-CD8- T cells had a higher percentage of cells taking up oligonucleotide than did B cells. T- or B-cell mitogens caused markedly increased oligonucleotide uptake in T or B cells, respectively. Oligonucleotide uptake could be inhibited only partially with competitor DNA. To distinguish between cell membrane-bound and intracellular oligonucleotide, cells were washed in acid glycine buffer (which removes most surface oligonucleotide). This demonstrated that most of the oligonucleotide was intracellular. We conclude that oligonucleotide uptake is quite heterogeneous among cultured cells, and that this uptake is inducible by mitogens. These data may be important for the design and interpretation of in vitro experiments, and for the planning of in vivo therapy with antisense oligonucleotides.
Krieg
A M
AM
Cellular Immunology Section, National Institute of Arthritis and Musculoskeletal and Skin Diseases, National Institutes of Health, Bethesda, MD 20892.
Gmelig-Meyling
F
F
Gourley
M F
MF
Kisch
W J
WJ
Chrisey
L A
LA
Steinberg
A D
AD
D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology / Q000378:metabolism*; D001483:Base Sequence; D001692:Biological Transport; D015704:CD4 Antigens / Q000032:analysis; D016827:CD8 Antigens / Q000032:analysis; D002478:Cells, Cultured; D007700:Kinetics; D008198:Lymph Nodes / Q000378:metabolism; D008213:Lymphocyte Activation; D051379:Mice; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000378:metabolism*; D013154:Spleen / Q000378:metabolism; D016176:T-Lymphocyte Subsets / Q000378:metabolism; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism*
D015704:CD4 Antigens; D016827:CD8 Antigens; D009838:Oligodeoxyribonucleotides