CLONING OF PORCINE INTERCELLULAR ADHESION MOLECULE-1 AND CHARACTERIZATION OF ITS INDUCTION ON ENDOTHELIAL CELLS BY CYTOKINES1

Stocker, Claire J.2; Sugars, Katharine L.2; Yarwood, Helen2; Delikouras, Alex3; Lechler, Robert I.3; Dorling, Anthony3; Landis, R. Clive2; Morley, Bernard J.3; Haskard, Dorian O.2 4

Experimental Transplantation

Background. The transplantation of pig organs into humans requires a detailed knowledge of similarities and differences between the two species in the molecular physiology of host defense mechanisms. We therefore set out to identify porcine intercellular adhesion molecule (ICAM)-1 and to characterize its expression by endothelial cells.

Methods. Porcine ICAM-1 cDNA was isolated from an endothelial cell cDNA library. An anti-pig ICAM-1 monoclonal antibody was generated and used to investigate the regulation by cytokines of ICAM-1 expression by porcine aortic endothelial cells (PAEC), using flow cytometry.

Results. We found that porcine ICAM-1 was similar in primary structure to human ICAM-1, with five Ig-like domains. COS-7 cells transfected with porcine ICAM-1 supported β2 but not α4 integrin-dependent adhesion of human T lymphoblasts. There was a low-level surface expression of ICAM-1 on unstimulated PAEC and increased expression after stimulation with tumor necrosis factor (TNF)-α. However expression of ICAM-1 seemed to be significantly lower than that of vascular cell adhesion molecule-1, both on unstimulated and TNF-α-activated PAEC. Recombinant porcine interferon-γ weakly stimulated ICAM-1 expression when incubated alone with PAEC but had an inhibitory effect on the increase in ICAM-1 due to TNF-α, both at 8 and 24 hr.

Conclusions. Our observations confirm the existence of ICAM-1 in the pig and provide novel insights into how porcine and human endothelial cells differ in terms of adhesion molecule expression and cytokine responsiveness. Such differences are potentially important in interpreting models of inflammation in the pig and also in understanding the process of rejection of porcine xenografts.

British Heart Foundation Cardiovascular Medicine Unit, National Heart and Lung Institute, and Department of Medicine, Imperial College School of Medicine, Hammersmith Hospital, London W12 0NN, United Kingdom

2 British Heart Foundation Cardiovascular Medicine Unit, National Heart and Lung Institute.

3 Department of Medicine, Imperial College School of Medicine, Hammersmith Hospital.

5 Genbank Accession number AF156712 (File Bankit 268211).

Received 28 September 1999.

Accepted 8 March 2000.

4 Address correspondence to: Dr. Dorian O. Haskard, BHF Cardiovascular Medicine Unit, NHLI, Imperial College School of Medicine, Hammersmith Hospital, London W12 ONN, UK. E-mail: d.haskard@ic.ac.uk.

1 This study was supported by a grant from the British Heart Foundation.

© 2000 Lippincott Williams & Wilkins, Inc.