Single-stranded polyinosinic acid oligonucleotides trigger leukocyte production of proteins belonging to fibrinolytic and coagulation cascades.
The present study assessed the inductory effects of ds- and ssRNA on the leukocyte production of proteins belonging to fibrinolytic and coagulation cascades. Murine splenocytes were stimulated with dsRNA [polyinosinic:polycytidylic acid (polyIC)] and ssRNA sequences [polyinosinic acid (polyI), polycytidylic acid (polyC), and polyuridylic acid (polyU)]. The expression of plasminogen (Plg), tissue factor (TF), IL-6, and IFN-alpha was assessed. Intracellular transduction mechanisms activated by oligonucleotides were evaluated using specific inhibitors of signaling pathways and genetically modified mice. polyIC efficiently and dose-dependently induced the expression of Plg, IL-6, and IFN-alpha, whereas TF was not induced by polyIC. polyI was unable to trigger IFN-alpha production, and it was efficiently inducing Plg and TF. IFN-alphaR and dsRNA-dependent protein kinase signaling were not required for the polyI-induced production of Plg or TF. Neither polyU nor polyC induced the expression of Plg or TF. Importantly, the presence of U- and C-nucleotide strands in the dsRNA significantly reduced expression of Plg and TF compared with polyI alone. Exposure of splenocytes to polyI activated the NF-kappaB pathway followed by the expression of TF and IL-6. In contrast, Plg production did not require NF-kappaB, was only partly down-regulated by p38 MAPK inhibitor, and was efficiently inhibited by insulin, indicating a different mechanism for its induction. ssRNA exerts its TF-generating properties through NF-kappaB activation in an IFN-alpha-independent manner. The expression of fibrinolytic versus coagulation proteins is regulated through distinctly different transduction pathways. As fibrinolytic and coagulation cascades are important components of inflammatory homeostasis, these findings might have importance for development of new, targeted therapies.
Department of Rheumatology and Inflammation Research, University of Göteborg, Sweden.
Fariba
F
Zare
Mattias
M
Magnusson
Linda Nilsson
LN
Möllers
Tao
T
Jin
Andrej
A
Tarkowski
Maria
M
Bokarewa
D000818:Animals; D001777:Blood Coagulation / Q000187:drug effects* / Q000502:physiology; D002478:Cells, Cultured; D005342:Fibrinolysis / Q000187:drug effects* / Q000502:physiology; D007328:Insulin / Q000494:pharmacology; D007369:Interferon Inducers / Q000494:pharmacology*; D016898:Interferon-alpha / Q000378:metabolism; D015850:Interleukin-6 / Q000235:genetics / Q000378:metabolism; D007962:Leukocytes / Q000187:drug effects* / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D053594:Myeloid Differentiation Factor 88 / Q000502:physiology; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D011070:Poly I-C / Q000494:pharmacology*; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D012330:RNA, Double-Stranded / Q000494:pharmacology; D017471:Receptors, Interferon / Q000502:physiology; D017472:Receptors, Interleukin-1 / Q000502:physiology; D015398:Signal Transduction / Q000187:drug effects*; D013154:Spleen / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D013925:Thromboplastin / Q000235:genetics / Q000378:metabolism; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism
D007328:Insulin; D007369:Interferon Inducers; D016898:Interferon-alpha; D015850:Interleukin-6; C507780:Myd88 protein, mouse; D053594:Myeloid Differentiation Factor 88; D016328:NF-kappa B; D011506:Proteins; D012330:RNA, Double-Stranded; D017471:Receptors, Interferon; D017472:Receptors, Interleukin-1; D013925:Thromboplastin; D048051:p38 Mitogen-Activated Protein Kinases; D011070:Poly I-C