Comparison of efficacy of antisense oligomers directed toward TNF-alpha in helper T and macrophage cell lines.
The authors investigated the use of antisense oligomers specific for TNF-alpha (AS-2) and nonsense control oligomers (NS) in T cells (HT2) and macrophages (RAW264.7), comparing three distinct chemical formulations. Phosphorothioate antisense (S-AS) caused sequence-specific inhibition of TNF-alpha production by activated HT2s (0.5 microM S-AS 2 vs S-NS: 31.4 +/- 1.2%, 4.2 +/- 3.2% inhibition, respectively). In contrast, S-AS were ineffective in RAW264.7, despite greater uptake as measured with fluorescent S-oligonucleotides. Furthermore, differences in efficacy of S-AS (HT2 > RAW) were not attributable to differences in the pinocytic (HT2 = RAW) or adsorptive endocytic (RAW > HT2) pathways implicated in oligonucleotide uptake, suggesting an important role for intracellular events after antisense uptake. Morpholino oligomers (M-AS), in contrast, were more effective in RAW264.7 than in HT2 (32.6 +/- 2.6% vs 12.3 +/- 0.5% inhibition), consistent with uptake experiments using fluorescent M-oligomers. Phosphodiester oligonucleotides were ineffective in both cell types. It was concluded that antisense efficacy in leukocytes varies according to type of oligomer, cell target and intracellular processing event(s).
Taylor
M F
MF
Physiology Program, Harvard School of Public Health, Boston, MA 02115, USA.
Paulauskis
J D
JD
Weller
D D
DD
Kobzik
L
L
D000818:Animals; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D004705:Endocytosis / Q000187:drug effects; D005434:Flow Cytometry; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D009025:Morpholines; D016376:Oligonucleotides, Antisense / Q000493:pharmacokinetics / Q000494:pharmacology*; D010710:Phosphates; D010714:Phosphatidylethanolamines / Q000494:pharmacology; D011069:Poly I / Q000494:pharmacology; D006377:T-Lymphocytes, Helper-Inducer / Q000187:drug effects / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013873:Thionucleotides; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis*
D009025:Morpholines; D016376:Oligonucleotides, Antisense; D010710:Phosphates; D010714:Phosphatidylethanolamines; D013873:Thionucleotides; D014409:Tumor Necrosis Factor-alpha; D011069:Poly I; C020888:1,2-dielaidoylphosphatidylethanolamine; D013755:Tetradecanoylphorbol Acetate