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

Torstein Jensen; Peter Kierulf; Per M. Sandset; Olav Klingenberg; Gunn B. Joø; Hans C. Godal; Ole H. Skjønsberg

doi:10.1160/TH07-01-0039

Thrombosis and Haemostasis, Table of Contents

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

¹Department of Haematology

²Department of Pulmonary Medicine

⁵Faculty Division Ullevål University Hospital, University of Oslo, Norway

,

Peter Kierulf

³Department of Clinical Chemistry, Ullevål University Hospital, Oslo, Norway

⁵Faculty Division Ullevål University Hospital, University of Oslo, Norway

,

Per M. Sandset

¹Department of Haematology

⁵Faculty Division Ullevål University Hospital, University of Oslo, Norway

,

Olav Klingenberg

⁴Department of Medical Biochemistry, Rikshospitalet-Radiumhospitalet Medical Center, Oslo, Norway

,

Gunn B. Joø

³Department of Clinical Chemistry, Ullevål University Hospital, Oslo, Norway

,

Hans C. Godal^†

¹Department of Haematology

,

Ole H. Skjønsberg

²Department of Pulmonary Medicine

⁵Faculty Division Ullevål University Hospital, University of Oslo, Norway

› Author Affiliations

Abstract

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

Full Text

References

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