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Thromb Haemost 2002; 88(05): 788-793
DOI: 10.1055/s-0037-1613303
Review Article
Schattauer GmbH

Phosphorotioated Oligonucleotides Trigger Synthesis of Human Coagulation Serine Proteases

Maria I. Bokarewa
1   Dept. of Rheumatology and Inflammation Research and Dept. of Clinical Virology, Sahlgrenska University Hospital, Göteborg, Sweden
,
Kristoffer Hellstrand
1   Dept. of Rheumatology and Inflammation Research and Dept. of Clinical Virology, Sahlgrenska University Hospital, Göteborg, Sweden
,
Andrej Tarkowski
1   Dept. of Rheumatology and Inflammation Research and Dept. of Clinical Virology, Sahlgrenska University Hospital, Göteborg, Sweden
› Author Affiliations Financial support The work has been supported by the Göteborg Medical Society, the Swedish Association Against Rheumatism, the King Gustaf V:s Foundation, the Swedish Medical Research Council, Cancerfonden, the Nanna Svartz’ Foundation, the Börje Dahlins Foundation, National Inflammation Network and the University of Göteborg
Further Information

Publication History

Received 19 November 2001

Accepted after resubmission 24 July 2002

Publication Date:
08 December 2017 (online)

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Summary

Rationale

CpG containing phosphorotioated oligonucleotides (ODN) are efficient adjuvants able to enhance macrophage and B cell activities. Their impact in the generation of coagulation and fibrinolytic factors has not been analysed.

Objectives

Production of coagulation and fibrinolytic proteins by human peripheral blood mononuclear cells (PBMC) treated with ODN was assessed.

Findings

ODN induced in vitro generation of tissue factor (TF), thrombin and plasminogen, by PBMC. Synthesis of TF and thrombin occurred mostly in monocytes, while plasminogen was produced by both monocytic and lymphocytic cell populations. Generation of these proteins stimulated by CpG was totally blocked by cycloheximide, indicating the requirement of ongoing protein synthesis. Protein synthesis was equally pronounced at stimulation with cytosine-phosphate-guanosine (CpG)- and GpC- containing ODN, and dependend on the presence of the phosphorotioate moiety backbone in the ODN. Plasminogen, synthesized by monocytes and lymphocytes, was shown to be the primary product of ODN activation, leading subsequently to the expression of TF and thrombin generation.

Conclusions

Our findings should be taken into consideration when assessing advantages and drawbacks of immunotherapy and gene therapy.

Keywords

Tissue factor - plasminogen - oligonucleotides
 

  • References

  • 1 Manders P, Thomas R. Immunology of DNA vaccines: CpG motifs and antigen presentation. Inflamm Res 2000; 49: 199-205.
    CrossrefPubMedSearch in Google Scholar
  • 2 Krieg AM. The role of CpG motifs in innate immunity. Curr Opin Immunol 2000; 12: 35-43.
    CrossrefPubMedSearch in Google Scholar
  • 3 Sester DP, Naik S, Beasley SJ, Hume DA, Stacey KJ. Phosphorothioate backbone modification modulates macrophage activation by CpG DNA. J Immunol 2000; 165: 4165-73.
    CrossrefPubMedSearch in Google Scholar
  • 4 Pejler G, Lunderius C, Tomasini-Johansson B. Macrophages synthesize factor X and secrete factor X/Xa-containing prothrombinase activity into the surrounding medium. Thromb Haemost 2000; 84: 429-35.
    Thieme ConnectPubMedSearch in Google Scholar
  • 5 Giesen PL, Rauch U, Bohrmann B, Kling D, Roque M, Fallon JT, Badimon JJ, Himber J, Riederer MA, Nemerson Y. Blood-borne tissue factor: another view of thrombosis. Proc Natl Acad Sci USA 1999; 96: 2311-5.
    CrossrefPubMedSearch in Google Scholar
  • 6 Ritchie H, Booth NA. The distribution of the secreted and intracellular forms of plasminogen activator inhibitor 2 (PAI-2) in human peripheral blood monocytes is modulated by serum. Thromb Haemost 1998; 79: 813-7.
    Thieme ConnectPubMedSearch in Google Scholar
  • 7 Manchanda N, Schwartz BS. Lipopolysaccharide-induced modulation of human monocyte urokinase production and activity. J Immunol 1990; 145: 4174-80.
    PubMedSearch in Google Scholar
  • 8 Blasi F. The urokinase receptor. A cell surface, regulated chemokine. APMIS 1999; 107: 96-101.
    CrossrefPubMedSearch in Google Scholar
  • 9 Broze GJ. The tissue factor pathway of coagulation: factor VII, tissue factor, and tissue factor pathway inhibitor. In Bloom Aea. ed. Haemostasis and Thrombosis. Churchill Livingstone; 1994: 349-77.
    Search in Google Scholar
  • 10 Raza SL, Nehring LC, Shapiro SD, Cornelius LA. Proteinase-activated receptor-1 regulation of macrophage elastase (MMP-12) secretion by serine proteinases. J Biol Chem 2000; 275: 41243-50.
    CrossrefPubMedSearch in Google Scholar
  • 11 Altieri DC, Stamnes SJ. Protease-dependent T cell activation: ligation of effector cell protease receptor-1 (EPR-1) stimulates lymphocyte proliferation. Cell Immunol 1994; 155: 372-83.
    CrossrefPubMedSearch in Google Scholar
  • 12 Tsao BP, Fair DS, Curtiss LK, Edgington TS. Monocytes can be induced by lipopolysaccharide-triggered T lymphocytes to express functional factor VII/VIIa protease activity. J Exp Med 1984; 159: 1042-57.
    CrossrefPubMedSearch in Google Scholar
  • 13 Schmitt M, Harbeck N, Thomssen C, Wilhelm O, Magdolen V, Reuning U, Ulm K, Hofler H, Janicke F, Graeff H. Clinical impact of the plasminogen activation system in tumor invasion and metastasis: prognostic relevance and target for therapy. Thromb Haemost 1997; 78: 285-96.
    Thieme ConnectPubMedSearch in Google Scholar
  • 14 Del Rosso M, Fibbi G, Pucci M, Cerinic MM. Antisense oligonucleotides against the urokinase receptor: a therapeutic strategy for the control of cell invasion in rheumatoid arthritis and cancer. Clin Exp Rheumatol 1998; 16: 389-93.
    PubMedSearch in Google Scholar
  • 15 Bickerstaff AA, Xia D, Pelletier RP, Orosz CG. Mechanisms of graft acceptance: evidence that plasminogen activator controls donor-reactive delayed-type hypersensitivity responses in cardiac allograft acceptor mice. J Immunol 2000; 164: 5132-9.
    CrossrefPubMedSearch in Google Scholar
  • 16 Del Prete G, De Carli M, Lammel RM, D’Elios MM, Daniel KC, Giusti B, Abbate R, Romagnani S. Th1 and Th2 T-helper cells exert opposite regulatory effects on procoagulant activity and tissue factor production by human monocytes. Blood 1995; 86: 250-7.
    PubMedSearch in Google Scholar
  • 17 Hamilton JA, Whitty GA, Last K, Royston AK, Hart PH, Burgess DR. Interleukin-4 suppresses plasminogen activator inhibitor-2 formation in stimulated human monocytes. Blood 1992; 80: 121-5.
    PubMedSearch in Google Scholar
  • 18 Mellqvist UH, Hansson M, Brune M, Dahlgren C, Hermodsson S, Hellstrand K. Natural killer cell dysfunction and apoptosis induced by chronic myelogenous leukemia cells: role of reactive oxygen species and regulation by histamine. Blood 2000; 96: 1961-8.
    PubMedSearch in Google Scholar
  • 19 Hellstrand K, Asea A, Dahlgren C, Hermodsson S. Histaminergic regulation of NK cells. Role of monocyte-derived reactive oxygen metabolites. J Immunol 1994; 153: 4940-7.
    PubMedSearch in Google Scholar
  • 20 Liang H, Nishioka Y, Reich CF, Pisetsky DS, Lipsky PE. Activation of human B cells by phosphorothioate oligodeoxynucleotides. J Clin Invest 1996; 98: 1119-29.
    CrossrefPubMedSearch in Google Scholar
  • 21 Hartmann G, Krieg AM. Mechanism and function of a newly identified CpG DNA motif in human primary B cells. J Immunol 2000; 164: 944-53.
    CrossrefPubMedSearch in Google Scholar
  • 22 Kandimalla ER, Manning A, Zhao Q, Shaw DR, Byrn RA, Sasisekharan V, Agrawal S. Mixed backbone antisense oligonucleotides: design, biochemical and biological properties of oligonucleotides containing 2’-5’-riboand 3’-5’-deoxyribonucleotide segments. Nucleic Acids Res 1997; 25: 370-8.
    CrossrefPubMedSearch in Google Scholar
  • 23 Henry SP, Novotny W, Leeds J, Auletta C, Kornbrust DJ. Inhibition of coagulation by a phosphorothioate oligonucleotide. Antisense Nucleic Acid Drug Dev 1997; 07: 503-10.
    CrossrefPubMedSearch in Google Scholar
  • 24 Deng GM, Tarkowski A. The features of arthritis induced by CpG motifs in bacterial DNA. Arthritis Rheum 2000; 43: 356-64.
    CrossrefPubMedSearch in Google Scholar
  • 25 Chiaramonte MG, Hesse M, Cheever AW, Wynn TA. CpG oligonucleotides can prophylactically immunize against Th2-mediated schistosome egg-induced pathology by an IL-12-independent mechanism. J Immunol 2000; 164: 973-85.
    CrossrefPubMedSearch in Google Scholar
  • 26 Varisco PA, Peclat V, van Ness K, Bischof-Delaloye A, So A, Busso N. Effect of thrombin inhibition on synovial inflammation in antigen induced arthritis. Ann Rheum Dis 2000; 59: 781-7.
    CrossrefPubMedSearch in Google Scholar
  • 27 Ploplis VA, French EL, Carmeliet P, Collen D, Plow EF. Plasminogen deficiency differentially affects recruitment of inflammatory cell populations in mice. Blood 1998; 91: 2005-9.
    PubMedSearch in Google Scholar
  • 28 Gunningham SP, Currie MJ, Han C, Robinson BA, Scott PA, Harris AL, Fox SB. The short form of the alternatively spliced flt-4 but not its ligand vascular endothelial growth factor C is related to lymph node metastasis in human breast cancers. Clin Cancer Res 2000; 06: 4278-86.
    PubMedSearch in Google Scholar
  • 29 Yang JL, Seetoo D, Wang Y, Ranson M, Berney CR, Ham JM, Russell PJ, Crowe PJ. Urokinase-type plasminogen activator and its receptor in colorectal cancer: independent prognostic factors of metastasis and cancerspecific survival and potential therapeutic targets. Int J Cancer 2000; 89: 431-9.
    CrossrefPubMedSearch in Google Scholar
  • 30 Huang YQ, Li JJ, Karpatkin S. Thrombin inhibits tumor cell growth in association with up-regulation of p21(waf/cip1) and caspases via a p53- independent, STAT-1-dependent pathway. J Biol Chem 2000; 275: 6462-8.
    CrossrefPubMedSearch in Google Scholar
  • 31 Cunningham MA, Romas P, Hutchinson P, Holdsworth SR, Tipping PG. Tissue factor and factor VIIa receptor/ligand interactions induce proinflammatory effects in macrophages. Blood 1999; 94: 3413-20.
    PubMedSearch in Google Scholar
  • 32 Senden NH, Jeunhomme TM, Heemskerk JW, Wagenvoord R, van’t Veer C, Hemker HC, Buurman WA. Factor Xa induces cytokine production and expression of adhesion molecules by human umbilical vein endothelial cells. J Immunol 1998; 161: 4318-24.
    PubMedSearch in Google Scholar
  • 33 Ruan QR, Zhang WJ, Hufnagl P, Kaun C, Binder BR, Wojta J. Anisodamine counteracts lipopolysaccharide-induced tissue factor and plasminogen activator inhibitor-1 expression in human endothelial cells: contribution of the NF-kappa b pathway. J Vasc Res 2001; 38: 13-9.
    CrossrefPubMedSearch in Google Scholar
  • 34 Oeth P, Parry GC, Mackman N. Regulation of the tissue factor gene in human monocytic cells. Role of AP-1, NF-kappa B/Rel, and Sp1 proteins in uninduced and lipopolysaccharide-induced expression. Arterioscler Thromb Vasc Biol 1997; 17: 365-74.
    CrossrefPubMedSearch in Google Scholar
  • 35 Edmead C, Kanthou C, Benzakour O. Thrombin activates transcription factors sp1, NF-kappaB, and CREB: importance of the use of phosphatase inhibitors during nuclear protein extraction for the assessment of transcription factor DNA-binding activities. Anal Biochem 1999; 275: 180-6.
    CrossrefPubMedSearch in Google Scholar