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Gut 47:88-96 doi:10.1136/gut.47.1.88
  • Inflammation and inflammatory bowel disease

Heparin attenuates TNF-α induced inflammatory response through a CD11b dependent mechanism

Abstract

BACKGROUND In addition to its anticoagulant properties, heparin has anti-inflammatory effects, the molecular and mechanistic bases of which are incompletely defined.

AIMS The current studies were designed to test the hypothesis that heparin abrogates the expression or function of leucocyte-endothelial adherence molecules which are fundamental to the acute inflammatory response.

METHODS The effects of heparin on tumour necrosis factor alpha (TNF-α) induced leucocyte rolling, adhesion, and migration as well as vascular permeability were assessed in rat mesenteric venules using intravital microscopy. Expression of adhesion molecules was quantitated using a double radiolabelled monoclonal antibody (mAb) binding technique in vivo (P-selectin, intercellular cell adhesion molecule type 1 (ICAM-1), and vascular cell adhesion molecule 1 (VCAM-1)) or flow cytometry (CD11a, CD11b, and L-selectin). Ex vivo binding of heparin to neutrophils was assessed by flow cytometry.

RESULTS TNF-α induced a significant increase in leucocyte rolling, adhesion, and migration, and vascular permeability, coincident with a significant increase in expression of P-selectin, ICAM-1, and VCAM-1. Ex vivo assessment of blood neutrophils showed significant upregulation of CD11a and CD11b and significant downregulation of L-selectin within five hours of TNF-α administration. Heparin pretreatment significantly attenuated leucocyte rolling, adhesion, and migration but did not affect expression of cell adhesion molecules or vascular permeability elicited by TNF-α administration. Binding of heparin was significantly increased on blood neutrophils obtained five hours after TNF-α administration. Preincubation with an anti-CD11b mAb but not with an anti-CD11a or anti-L-selectin antibody significantly diminished heparin binding ex vivo.

CONCLUSIONS Our results support the concept that the anti-inflammatory effects of heparin involve attenuation of a CD11b dependent adherent mechanism.

Footnotes

  • Abbreviations used in this paper:
    aPTT
    partial thromboplastin time
    BBS
    bicarbonate buffered saline
    BSA
    bovine serum albumin
    FITC
    fluorescein isothiocyanate
    fMLP
    N-formyl methionyl leucyl phenylalanine
    IBD
    inflammatory bowel disease
    ICAM-1
    intercellular cell adhesion molecule type 1
    Ig
    immunoglobulin
    IL-8
    interleukin 8
    mAb
    monoclonal antibody
    PBS
    phosphate buffered saline
    TNF-α
    tumour necrosis factor alpha
    VCAM-1
    vascular cell adhesion molecule 1
    PE
    phycoerythrin