Soluble ICAM-1 (sICAM-1) provokes PMN elastase release

J Surg Res. 1996 Jun;63(1):6-10. doi: 10.1006/jsre.1996.0213.

Abstract

Elevated levels of soluble intercellular adhesion molecule-1 have been shown predictive of post-injury multiple organ failure. We hypothesized that sICAM-1 augments distant organ injury via its affect on the PMN and; thus, have examined neutrophil elastase and superoxide production in response to sICAM-1. To obtain soluble ICAM-1, Chinese Hamster Ovarian (CHO) cells were transfected with human ICAM-1 (cDNA vector CD1.8), lysed and centrifuged at 150,000g for 1 hr; supernatant was passed over an ICAM-1 affinity gradient, eluted with 0.1 mM glycine x HCl, and concentrated using an Amicon Spin-X filter. PMNs were incubated for 1 hr with sICAM-1 at 37 degrees C. Quiescent and PMA-activated PMNs served as negative and positive controls respectively. Elastase activity was measured by the cleavage of methoxy-succinyl-alalyl-alalyl-prolyl-valyl-p-nitroanilide. Superoxide production was determined by superoxide dismutase inhibitive ferricytochrome C reduction over a 5-60 min incubation. PMN incubation with sICAM-1 provoked marked increase in elastase release 10.43 +/- 2.90 (10(-6) U/hr) compared to control 1.64 +/- 0.57, and was equivalent to PMA-activated PMN elastase release 11.60 +/- 1.50 (10(-6) U/hr). In contrast, sICAM-1 alone did not promote spontaneous PMN superoxide production beyond buffer treated PMNs (0.25 +/- 0.09 nmole/2.5 x 10(5) PMN/min). In sum, sICAM-1 stimulates PMN elastase release in vitro. Clinically, this may represent a mechanism by which sICAM-1 participates in the genesis of post-injury multiple organ failure.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • CHO Cells
  • Chromatography, Affinity
  • Cricetinae
  • Endothelium, Vascular / physiology
  • Humans
  • In Vitro Techniques
  • Intercellular Adhesion Molecule-1 / biosynthesis
  • Intercellular Adhesion Molecule-1 / isolation & purification
  • Intercellular Adhesion Molecule-1 / pharmacology*
  • Leukocyte Elastase
  • Models, Biological
  • Molecular Sequence Data
  • Neutrophils / drug effects
  • Neutrophils / enzymology*
  • Pancreatic Elastase / metabolism*
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / pharmacology
  • Substrate Specificity
  • Superoxides / blood
  • Tetradecanoylphorbol Acetate / pharmacology
  • Ultrafiltration

Substances

  • Recombinant Proteins
  • Superoxides
  • Intercellular Adhesion Molecule-1
  • Pancreatic Elastase
  • Leukocyte Elastase
  • Tetradecanoylphorbol Acetate