Transforming growth factor-beta-mediated regulation of human peripheral blood mononuclear cell proliferation as detected with phosphorothioate antisense oligodeoxynucleotides.
Transforming growth factor-beta (TGF-beta) is a potent cytokine that has influence upon immunosuppressive as well as proinflammatory processes. In this context we have investigated the role of endogenous TGF-beta in human peripheral blood mononuclear cells (PBMCs) by TGF-beta 1 phosphorothioate antisense oligodeoxynucleotides (TGF-beta 1-S-ODNs). In short-term cultures (up to 3 days), TGF-beta 1-S-ODNs-treated, interleukin 2-activated PBMCs displayed a growth advantage (up to 148%) compared to nonsense or untreated controls as measured by cell counting and [3H]thymidine incorporation. Long-term antagonization of TGF-beta production (up to 12 days) showed no significant differences between both antisense- and nonsense-treated PBMCs. The efficacy and specificity of TGF-beta 1-S-ODNs effects was validated by gene expression studies (Northern blot and ELISA) and analysis of cellular uptake of BrdU-labeled S-ODN (Immunocytochemistry). TGF-beta 1-S-ODNs at the final concentration of 1 microM reduced TGF-beta 1 protein concentration up to 65% in PBMCs cultures without significant changes in TGF-beta 1 mRNA expression. These experiments demonstrate that TGF-beta 1-S-ODNs specifically antagonize TGF-beta 1-mediated autocrine suppression of IL-2-dependent PBMC activation. TGF-beta 1-specific antisense oligonucleotides may represent a promising immunoresponse modifying agent that could be used for T-cell directed immunostimulation.
Jachimczak
P
P
Department of Neurology, University of Würzburg, Germany.
Fabel-Schulte
K
K
Hessdörfer
B
B
Brysch
W
W
Schlingensiepen
K H
KH
Blesch
A
A
Bogdahn
U
U
D001483:Base Sequence; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000166:cytology*; D008969:Molecular Sequence Data; D016376:Oligonucleotides, Antisense / Q000378:metabolism / Q000494:pharmacology*; D012333:RNA, Messenger / Q000032:analysis; D013873:Thionucleotides / Q000378:metabolism / Q000494:pharmacology*; D013997:Time Factors; D016212:Transforming Growth Factor beta / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology*
D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; D013873:Thionucleotides; D016212:Transforming Growth Factor beta