Characterization in vitro and in vivo of the pig analogue of human CD59 using new monoclonal antibodies.
CD59 is the sole characterized regulator of the complement membrane attack complex in humans. It is very widely and abundantly distributed, being present on all circulating cells, endothelia and epithelia, and in most tissues. CD59 analogues in rodents are distributed similarly. Interest in complement regulation in the pig has developed out of the current enthusiasm to exploit this species as a donor in xenotransplantation of organs to humans. We have recently isolated and cloned the pig analogue of human CD59. We here report the development and characterization of monoclonal antibodies against pig CD59. We have used these antibodies to develop efficient methods for the purification of pig CD59 to homogeneity from erythrocyte membranes and have obtained new information on the structure and function of the purified protein. The antibodies were found to function well in immunohistochemistry and have been used to perform a comprehensive survey of the expression and distribution of pig CD59 on cells and in organs of normal pigs. Pig CD59, like human CD59, is broadly expressed but there are some striking differences in tissue distribution, notably the apparent lack of pig CD59 on circulating platelets and on a subset of leucocytes in blood and lymphoid organs. The reported findings have important implications for the current approaches to avoiding complement-mediated hyperacute rejection in pig-to-human xenografts.
Hanna
S M
SM
Department of Medical Biochemistry, University of Wales College of Medicine, Cardiff, UK.
Williams
G T
GT
Van Den Berg
C W
CW
Morgan
B P
BP
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D000943:Antigens, Differentiation / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D001667:Binding, Competitive; D018957:CD59 Antigens / Q000276:immunology*; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D005434:Flow Cytometry; D006801:Humans; D007124:Immunoenzyme Techniques; D013045:Species Specificity; D013552:Swine / Q000276:immunology*; D014018:Tissue Distribution
D000911:Antibodies, Monoclonal; D000943:Antigens, Differentiation; D018957:CD59 Antigens