Biophysical characteristics of anti-Gal(alpha)1-3Gal IgM binding to cell surfaces: implications for xenotransplantation.
BACKGROUND
Natural antibodies directed against cell surface carbohydrates are thought to be vital to host defense and to initiate the rejection of xenografts and ABO-incompatible allografts. The biophysical properties underlying the association and dissociation of these antibodies from cell surfaces is incompletely understood. We investigated those properties for the binding of Galalpha1-3Gal antibodies to porcine endothelial cell surfaces, because such interactions might be relevant to the clinical application of xenotransplantation.


CONCLUSIONS
The initial rate of binding of anti-Galalpha1-3Gal antibodies to endothelial cells was found to depend on antibody concentration, antibody diffusion, and antigen concentration. The presence of an intact glycocalyx had a greater impact on antibody binding than mobility of antigen in cell membranes. Disruption of glycocalyx increased the amount of antibody bound at equilibrium by more than 50%. Although the binding of anti-Galalpha1-3Gal antibodies to cell surfaces could be inhibited by soluble Galalpha1-3Gal, once bound, some anti-Galalpha1-3Gal could not be dissociated by competitive inhibitors of binding or by denaturation of the bound Ig with chaotropic reagents, but could be dissociated by reduction of disulfide bonds, suggesting that attachment to cell surfaces was, at least in part, by means other than specific reaction with the epitope.
Parker
W
W
Department of Surgery, Duke University Medical Center, Durham, North Carolina 27710, USA.
Stitzenberg
K B
KB
Yu
P B
PB
Pratt
V S
VS
Nakamura
Y C
YC
Farel
L S
LS
Ward
C M
CM
Lin
S S
SS
Everett
M L
ML
Platt
J L
JL
D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000737:chemistry* / Q000378:metabolism*; D004187:Disaccharides / Q000276:immunology*; D000939:Epitopes / Q000737:chemistry; D019276:Glycocalyx / Q000502:physiology; D006801:Humans; D011485:Protein Binding; D012680:Sensitivity and Specificity; D013552:Swine; D013696:Temperature; D014183:Transplantation, Heterologous / Q000502:physiology; D014783:Viscosity
D000888:Antibodies, Anti-Idiotypic; D004187:Disaccharides; D000939:Epitopes; C044203:anti-IgM; C055075:galactosyl-(1-3)galactose