title: An investigation of the specificity of induced anti-pig antibodies in baboons.
abstract: OBJECTIVE
  To provide information on the specificity of induced anti-pig antibodies (Abs) in baboons after exposure and sensitization to pig antigens.
  
  
  METHODS
  Baboons (n=7) received either porcine mobilized peripheral blood progenitor cells (n=3), kidney (n=3) or heart (n=1) transplants. After rejection of these cells or organs, pre- and post-rejection sera were analyzed by enzyme-linked immunosorbent assay (ELISA) and flow cytometry to detect and measure anti-Galactosealpha1,3Galactose (Gal) and anti-non-Gal Abs. To study the anti-non-Gal carbohydrate response, the sera were incubated with pig red blood cells pretreated with alpha-galactosidase (to remove Gal) and three other exoglycosidases to remove other potential oligosaccharide epitopes, and studied by flow cytometry. To study the anti-swine leukocyte antigen (SLA) response, non-Gal Abs from two baboons sensitized with kidneys from inbred miniature swine of dd or aa haplotype, respectively, were adsorbed on cells of aa, cc, or dd haplotypes, and binding to aa, cc or dd cells was measured by flow cytometry. Cytotoxicity of anti-non-Gal Abs was tested in vitro by a complement-mediated cytotoxicity assay, using pig cells as targets.
  
  
  RESULTS
  In pre-transplant and pre-rejection sera, anti-Gal Abs were detected, but anti-non-Gal Abs were either absent or at minimal levels. After exposure to pig antigens, baboons developed induced anti-Gal and anti-non-Gal Abs. No anti-non-Gal Abs directed to the tested carbohydrate epitopes could be detected. Anti-non-Gal Abs showed minor evidence of specific SLA haplotype reactivity, suggesting that the major Ab response was to pan-pig determinants. Anti-non-Gal Abs showed a low level of complement-mediated lysis of pig cells in vitro.
  
  
  CONCLUSIONS
  In this limited study, no Ab response to non-Gal carbohydrates was observed, and anti-SLA specificity was minor, indicating that most induced anti-non-Gal Ab was directed against non-specific pig proteins, including SLA-epitopes.
authors.affiliation: The Transplantation Biology Research Center, Massachusetts General Hospital/Harvard Medical School, Boston, MA, USA.
authors.forename: L
authors.initials: L
authors.lastname: Buhler
authors.forename: Y
authors.initials: Y
authors.lastname: Xu
authors.forename: W
authors.initials: W
authors.lastname: Li
authors.forename: A
authors.initials: A
authors.lastname: Zhu
authors.forename: D K C
authors.initials: DK
authors.lastname: Cooper
mesh_terms: D000818:Animals; D000910:Antibodies, Heterophile / Q000276:immunology*; D000918:Antibody Specificity; D015478:Antigens, Heterophile / Q000276:immunology; D006084:Graft Rejection / Q000276:immunology; D016027:Heart Transplantation / Q000276:immunology*; D006596:Hexosaminidases / Q000276:immunology; D016030:Kidney Transplantation / Q000276:immunology*; D010215:Papio; D013552:Swine; D014183:Transplantation, Heterologous / Q000276:immunology*; D000519:alpha-Galactosidase / Q000276:immunology; D048809:alpha-N-Acetylgalactosaminidase; D001616:beta-Galactosidase / Q000276:immunology; D001619:beta-N-Acetylhexosaminidases / Q000276:immunology
chemical_list: D000910:Antibodies, Heterophile; D015478:Antigens, Heterophile; D006596:Hexosaminidases; D000519:alpha-Galactosidase; D001616:beta-Galactosidase; D048809:alpha-N-Acetylgalactosaminidase; D001619:beta-N-Acetylhexosaminidases