Download PDF
Thromb Haemost 2007; 97(05): 822-829
DOI: 10.1160/TH07-01-0039
Wound Healing and Inflammation / Infection
Schattauer GmbH

Fibrinogen and fibrin induce synthesis of proinflammatory cytokines from isolated peripheral blood mononuclear cells

Torstein Jensen
1   Department of Haematology
2   Department of Pulmonary Medicine
5   Faculty Division Ullevål University Hospital, University of Oslo, Norway
,
Peter Kierulf
3   Department of Clinical Chemistry, Ullevål University Hospital, Oslo, Norway
5   Faculty Division Ullevål University Hospital, University of Oslo, Norway
,
Per M. Sandset
1   Department of Haematology
5   Faculty Division Ullevål University Hospital, University of Oslo, Norway
,
Olav Klingenberg
4   Department of Medical Biochemistry, Rikshospitalet-Radiumhospitalet Medical Center, Oslo, Norway
,
Gunn B. Joø
3   Department of Clinical Chemistry, Ullevål University Hospital, Oslo, Norway
,
Hans C. Godal   †
1   Department of Haematology
,
Ole H. Skjønsberg
2   Department of Pulmonary Medicine
5   Faculty Division Ullevål University Hospital, University of Oslo, Norway
› Author Affiliations
Further Information

Publication History

Received 20 January 2007

Accepted after revision 02 March 2007

Publication Date:
24 November 2017 (online)

Also available at
    Permissions and Reprints

Summary

Fibrinogen in plasma includes three main fractions; high-molecular- weight (HMW) -fibrinogen, low-molecular-weight (LMW) -fibrinogen, and very-low-molecular-weight (LMW`) -fibrinogen. During acute-phase conditions, plasma fibrinogen levels and the HMW-/LMW-fibrinogen ratio increase rapidly due to increased synthesis of HMW-fibrinogen. The consequences of elevated plasma fibrinogen levels and local deposition of fibrin in inflammatory tissues observed during acute-phase conditions are not clear.We wanted to investigate proinflammatory effects of fibrinogen and fibrin on peripheral blood mononuclear cells (PBMC) as reflected by altered mRNA expression and synthesis of the proinflammatory cytokines IL-6,TNF- α and IL-1 β, and to explore the significance of altered HMW-/LMW-fibrinogen ratio. PBMC were isolated from whole blood using Lymphoprep® . HMW-fibrinogen was separated from unfractioned fibrinogen by ammonium sulphate precipitation. Cells were incubated with unfractioned fibrinogen, HMW-fibrinogen or fibrin. Cytokine levels in cell lysates were determined using ELISA assays. Real-time PCR was used for mRNA quantification. We found that fibrinogen significantly increased mRNA levels, and induced synthesis of the proinflammatory cytokines IL-6 and TNF- α in PBMC in a dose dependent manner. Median (25, 75 percentile) IL-6 and TNF- α concentrations were 12 (5, 40) pg/ ml and 16 (0,61) pg/ml,respectively.Median mRNA quantity was increased 12.3– (6.6, 48.6) and 1.7– (1.5, 6.5) fold for IL-6 and TNF- α compared to controls.The stimulatory effect of unfractioned fibrinogen was not significantly different from HMW-fibrinogen. Fibrinogen and fibrin were equally effective in promoting cytokine synthesis from PBMC.The results support that fibrin and fibrinogen may actively modulate the inflammatory process by inducing synthesis of proinflammatory cytokines from PBMC.

Keywords

Fibrinogen - fibrin - PMBC - inflammation - cytokines

† Deceased March 24, 2005


 

  • References

  • 1 Holm B, Godal HC.. Quantitation of the three normally- occurring plasma fibrinogens in health and during so-called "acute phase" by SDS electrophoresis of fibrin obtained from EDTA-plasma. Thromb Res 1984; 35: 279-290.
    CrossrefPubMedSearch in Google Scholar
  • 2 Drury D, McMaster P. The liver as the source of fibrinogen. J Exp Med 1929; 50: 569-678.
    CrossrefPubMedSearch in Google Scholar
  • 3 Lipinska I, Lipinski B, Gurewich V. et al. Fibrinogen heterogeneity in cancer, in occlusive vascular disease, and after surgical procedures. Am J Clin Pathol 1976; 66: 958-966.
    CrossrefPubMedSearch in Google Scholar
  • 4 Stone MC, Thorp JM. Plasma fibrinogen--a major coronary risk factor. J R Coll Gen Pract 1985; 35: 565-569.
    PubMedSearch in Google Scholar
  • 5 Wilhelmsen L, Svardsudd K, Korsan-Bengtsen K. et al. Fibrinogen as a risk factor for stroke and myocardial infarction. N Engl J Med 1984; 23 (311) 501-505.
    PubMedSearch in Google Scholar
  • 6 Meade TW, Mellows S, Brozovic M. et al. Haemostatic function and ischaemic heart disease: principal results of the Northwick Park Heart Study. Lancet 1991; 338: 1079
    PubMedSearch in Google Scholar
  • 7 Hoff HF, Heideman CL, Jackson RL. et al. Localization patterns of plasma apolipoproteins in human atherosclerotic lesions. Circ Res 1975; 37: 72-79.
    CrossrefPubMedSearch in Google Scholar
  • 8 Sadoshima S, Tanaka K. Fibrinogen and low density lipoprotein in the development of cerebral atherosclerosis. Atherosclerosis 1979; 34: 93-103.
    CrossrefPubMedSearch in Google Scholar
  • 9 Woolf N, Crawford T. Fatty streaks in the aortic intima studied by an immuno-histochemical technique. J Pathol Bacteriol 1960; 80: 405-408.
    CrossrefPubMedSearch in Google Scholar
  • 10 Perez RL, Roman J. Fibrin enhances the expression of IL-1 beta by human peripheral blood mononuclear cells. Implications in pulmonary inflammation. J Immunol 1995; 154: 1879-1887.
    PubMedSearch in Google Scholar
  • 11 Perez RL, Ritzenthaler JD, Roman J. Transcriptional regulation of the interleukin-1beta promoter via fibrinogen engagement of the CD18 integrin receptor. Am J Respir Cell Mol Biol 1999; 20: 1059-1066.
    CrossrefPubMedSearch in Google Scholar
  • 12 Dinarello CA. Proinflammatory cytokines. Chest 2000; 118: 503-508.
    CrossrefPubMedSearch in Google Scholar
  • 13 Altieri DC, Plescia J, Plow EF. The structural motif glycine 190-valine 202 of the fibrinogen gamma chain interacts with CD11b/CD18 integrin (alpha M beta 2, Mac-1) and promotes leukocyte adhesion. J Biol Chem 1993; 268: 1847-1853.
    PubMedSearch in Google Scholar
  • 14 Ugarova TP, Yakubenko VP. Recognition of fibrinogen by leukocyte integrins. Ann NY Acad Sci 2001; 936: 368-385.
    PubMedSearch in Google Scholar
  • 15 Graves KL, Roman J. Fibronectin modulates expression of interleukin-1 beta and its receptor antagonist in human mononuclear cells. Am J Physiol 1996; 271: L61-L69.
    CrossrefPubMedSearch in Google Scholar
  • 16 Leroyer C, Escoffre M, Le Moigne E. et al. Diagnostic value of a new sensitive membrane based technique for instantaneous D-dimer evaluation in patients with clinically suspected deep venous thrombosis. Thromb Haemost 1997; 77: 637-40.
    Thieme ConnectPubMedSearch in Google Scholar
  • 17 Jensen T, Halvorsen S, Godal HC. et al. The viscosity of fibrinogen subfractions and of EDTA denatured fibrinogen do not differ from that of native fibrinogen. Thromb Res 2004; 113: 51-56.
    CrossrefPubMedSearch in Google Scholar
  • 18 Weinstein MJ, Deykin D. Quantitative abnormality of an Aalpha chain molecular weight form in the fibrinogen of cirrhotic patients. Br J Haematol 1978; 40: 617-630.
    CrossrefPubMedSearch in Google Scholar
  • 19 Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 2001; 25: 402-408.
    CrossrefPubMedSearch in Google Scholar
  • 20 Jensen T, Halvorsen S, Godal HC. et al. Discrep ancy between fibrinogen concentrations determined by clotting rate and clottability assays during the acutephase reaction. Thromb Res 2000; 100: 397-403.
    CrossrefPubMedSearch in Google Scholar
  • 21 De Groote D, Zangerle PF, Gevaert Y. et al. Direct stimulation of cytokines (IL-1 beta, TNF-alpha, IL-6, IL-2, IFN-gamma and GM-CSF) in whole blood. I. Comparison with isolated PBMC stimulation. Cytokine 1992; 04: 239-248.
    CrossrefPubMedSearch in Google Scholar
  • 22 Oliver JC, Bland LA, Oettinger CW. et al. Cytokine kinetics in an in vitro whole blood model following an endotoxin challenge. Lymphokine Cytokine Res 1993; 12: 115-20.
    PubMedSearch in Google Scholar
  • 23 Sitrin RG, Pan PM, Srikanth S. et al. Fibrinogen activates NF-kappa B transcription factors in mononuclear phagocytes. J Immunol 1998; 161: 1462-1470.
    PubMedSearch in Google Scholar
  • 24 Altieri DC. Occupancy of CD11b/CD18 (Mac-1) divalent ion binding site(s) induces leukocyte adhesion. J Immunol 1991; 147: 1891-1898.
    PubMedSearch in Google Scholar
  • 25 Parrillo JE. Pathogenetic mechanisms of septic shock. N Engl J Med 1993; 328: 1471-1477.
    CrossrefPubMedSearch in Google Scholar
  • 26 Ruoslahti E. RGD and other recognition sequences for integrins. Annu Rev Cell Dev Biol 1996; 12: 697-715.
    CrossrefPubMedSearch in Google Scholar
  • 27 Dvorak HF, Senger DR, Dvorak AM. et al. Regulation of extravascular coagulation by microvascular permeability. Science 1985; 227: 1059-1061.
    CrossrefPubMedSearch in Google Scholar
  • 28 Clark RA, Lanigan JM, DellaPelle P. et al. Fibronectin and fibrin provide a provisional matrix for epidermal cell migration during wound reepithelialization. J Invest Dermatol 1982; 79: 264-269.
    CrossrefPubMedSearch in Google Scholar
  • 29 Fan ST, Edgington TS. Coupling of the adhesive receptor CD11b/CD18 to functional enhancement of effector macrophage tissue factor response. J Clin Invest 1991; 87: 50-57.
    CrossrefPubMedSearch in Google Scholar
  • 30 Fan ST, Edgington TS. Integrin regulation of leukocyte inflammatory functions. CD11b/CD18 enhancement of the tumor necrosis factor-alpha responses of monocytes. J Immunol 1993; 150: 2972-2980.
    PubMedSearch in Google Scholar
  • 31 Pakianathan DR. Extracellular matrix proteins and leukocyte function. J Leukoc Biol 1995; 57: 699-702.
    PubMedSearch in Google Scholar
  • 32 Trezzini C, Schuepp B, Maly FE. et al. Evidence that exposure to fibrinogen or to antibodies directed against Mac-1 (CD11b/CD18; CR3) modulates human monocyte effector functions. Br J Haematol 1991; 77: 16-24.
    CrossrefPubMedSearch in Google Scholar
  • 33 Kang YH, Lee CH, Monroy RL. et al. Uptake, distribution and fate of bacterial lipopolysaccharides in monocytes and macrophages: an ultrastructural and functional correlation. Electron Microscopy Rev 1992; 05: 381-419.
    PubMedSearch in Google Scholar
  • 34 Lin WJ, Yeh WC. Implication of Toll-like receptor and tumor necrosis factor alpha signaling in septic shock. Shock 2005; 24: 206-209.
    CrossrefPubMedSearch in Google Scholar
  • 35 Maris NA, de Vos AF, Bresser P. et al. Activation of coagulation and inhibition of fibrinolysis in the lung after inhalation of lipopolysaccharide by healthy volunteers. Thromb Haemost 2005; 93: 1036-40.
    Thieme ConnectPubMedSearch in Google Scholar
  • 36 Pan SL, Guh JH, Peng CY. et al. A potential role of YC-1 on the inhibition of cytokine release in peripheral blood mononuclear leukocytes and endotoxemic mouse models. Thromb Haemost 2005; 93: 940-948.
    Thieme ConnectPubMedSearch in Google Scholar
  • 37 Wu KH, Wu WM, Lu MY. et al. Inhibitory effect of pooled human immunoglobulin on cytokine production in peripheral blood mononuclear cells. Pediatr Allergy Immunol 2006; 17: 60-68.
    CrossrefPubMedSearch in Google Scholar
  • 38 Roman J, Ritzenthaler JD, Perez RL. et al. Differential modes of regulation of interleukin-1beta expression by extracellular matrices. Immunology 1999; 98: 228-237.
    CrossrefPubMedSearch in Google Scholar
  • 39 Tang L, Eaton JW.. et al. Fibrin(ogen) mediates acute inflammatory responses to biomaterials. J Exp Med 1993; 178: 2147-2156.
    CrossrefPubMedSearch in Google Scholar
  • 40 Tang L, Ugarova TP, Plow EF. et al. Molecular determinants of acute inflammatory responses to biomaterials. J Clin Invest 1996; 97: 1329-1334.
    CrossrefPubMedSearch in Google Scholar
  • 41 Busso N, Peclat V, Van Ness K. et al. Exacerbation of antigen-induced arthritis in urokinase-deficient mice. J Clin Invest 1998; 102: 41-50.
    CrossrefPubMedSearch in Google Scholar