Alpha-galactosyl epitope-mediated activation of porcine aortic endothelial cells: type I activation.
BACKGROUND
The galactose alpha(1-3)galactose (alpha-gal) epitope associated with membrane glycoproteins and glycolipids represents a major determinant recognized on porcine cells by human xenoreactive natural antibodies (XNA). Together, bound XNA and complement rapidly induce porcine aortic endothelial cell (PAEC) activation; this process is associated with cellular shape changes, transient development of intercellular gaps, and loss of ATDPase and thrombomodulin, with release of heparan sulfate. The aim of this study was to evaluate patterns of type I endothelial cell activation (i.e., activation that does not require protein synthesis) following ligation of alpha-gal epitopes with anti-Gal antibodies and alpha-gal-specific lectins.


RESULTS
PAEC incubated in the presence of the alpha-gal binding, Bandeiraea simplicifolia lectin (BS-I) underwent cellular shape changes associated with the formation of intercellular gaps. PAEC exposure to BS-I was also associated with the tyrosine phosphorylation of a protein (apparent molecular mass of approximately 130 kDa), not observed following lipopolysaccharide, tumor necrosis factor, or XNA stimulation. This lectin-induced tyrosine phosphorylation was not affected by cytochalasin D (inhibitor of actin filament polymerization), by genistein (inhibitor of tyrosine kinases), or by staurosporine (inhibitor of tyrosine phosphorylation and protein kinase C). In addition, incubation of PAEC with BS-I and monoclonal anti-Gal IgM induced p42/44 map kinase and activated the transcription factor NF-kappaB.


CONCLUSIONS
Agonist binding of alpha-gal can evoke endothelial cell activation independently of complement activation. These observations have implications for the survival of xenografts.
Palmetshofer
A
A
Novartis Center for Immunobiology, Department of Surgery, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts 02215, USA.
Galili
U
U
Dalmasso
A P
AP
Robson
S C
SC
Bach
F H
FH
D000199:Actins / Q000378:metabolism; D000818:Animals; D000954:Antigens, Surface / Q000276:immunology*; D001011:Aorta / Q000166:cytology / Q000276:immunology; D002478:Cells, Cultured; D003599:Cytoskeleton / Q000648:ultrastructure; D004187:Disaccharides / Q000276:immunology*; D004730:Endothelium, Vascular / Q000276:immunology*; D006001:Glycoconjugates / Q000276:immunology; D007365:Intercellular Junctions / Q000648:ultrastructure; D037102:Lectins / Q000276:immunology; D008297:Male; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism; D019000:Phosphotyrosine / Q000378:metabolism; D037121:Plant Lectins; D013552:Swine
D000199:Actins; D000954:Antigens, Surface; D004187:Disaccharides; D006001:Glycoconjugates; C045475:Griffonia simplicifolia lectins; D037102:Lectins; D016328:NF-kappa B; D037121:Plant Lectins; C055075:galactosyl-(1-3)galactose; D019000:Phosphotyrosine; D019950:Mitogen-Activated Protein Kinase 1