HIGH-LEVEL PORCINE ENDOTHELIAL CELL EXPRESSION OF ??(1,2)-FUCOSYLTRANSFERASE REDUCES HUMAN MONOCYTE ADHESION AND ACTIVATION1

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Abstract
Background. Monocyte binding to and activation by human endothelium requires a number of interactions, including those involving sialylated endothelial cell ligands. As porcine endothelial cell transfection with α(1,2)-fucosylatransferase has been shown to reduce terminal sialylation, we investigated whether high-level expression of α(1,2)-fucosyltransferase by porcine endothelium would reduce human monocyte adhesion and functional activation. Methods. Purified human monocytes were labeled with 51Cr, and measured for adherence to human or porcine endothelial cell monolayers in the presence of either medium or monoclonal antibodies against monocyte lectins or sialylated endothelial cell ligands. Monocyte production of prostaglandin E2 (PGE2) and interleukin-1β (IL-1β) was measured by enzyme-linked immunosorbent assay, using supernatants collected from cultures performed between human monocytes and human or porcine endothelial cell monolayers. Finally, monocyte adhesion and activation were measured after culture with a porcine endothelial cell line transfected with α(1,2)-fucosyltransferase, expressing reduced surface expression of terminal Gal α(1,3)-Gal and sialic acid residues. Results. Human monocytes adhered by 50% higher levels to porcine endothelium than to human endothelium. This increased level of adherence was associated with augmented monocyte activation, as defined by 3.3-fold higher levels of PGE2 production and 7.3-fold higher levels of IL-1β production. Monoclonal antibodies against CD62L (L-selectin) on monocytes or CD15s (sialyated Lewis X) on porcine endothelium reduced monocyte adhesion by 38% and 52%, respectively. Porcine endothelial cell transfection with α(1,2)-fucosyltransferase reduced terminal sialic acid expression by 65%, monocyte adherence by 50%, and the production of PGE2 and IL-1β by 67% and 38%, respectively. Conclusions. Together, there results demonstrate that human monocytes use surface lectins to bind to sialylated carbohydrate structures on porcine endothelium, and indicate that reduction in porcine endothelial cell surface expression of terminally sialylated structures by high-level α(1,2)-fucosyltransferase activity reduces monocyte adherence and activation.