M Tanemura1, S Maruyama, U Galili. 1. Department of Microbiology and Immunology, MCP Hahnemann School of Medicine, Philadelphia, Pennsylvania 19129, USA.
Abstract
BACKGROUND: Expression of the alpha-gal epitope in mice can be completely eliminated by disruption of the alpha1,3 galactosyltransferase gene. As an initial step for assessing the feasibility of this approach in the pig, it was of interest to compare the expression of alpha-gal epitopes in pig and mouse organs. METHODS: Membranes from pig and mouse organ homogenates were analyzed for alpha-gal epitope expression by Western blots, enzyme-linked immunosorbent assay (ELISA), immunostaining of tissues, and ELISA inhibition assay. RESULTS: Immunostaining of Western blots with human anti-Gal detected alpha-gal epitopes on glycoproteins from pig organs but not on glycoproteins from the corresponding mouse organs. ELISA with membrane homogenates and immunostaining of tissue sections demonstrated a much higher binding of human anti-Gal to alpha-gal epitopes on pig membranes than on mouse membranes. ELISA inhibition assay with monoclonal anti-Gal indicated that alpha-gal epitope expression in pig organs is up to 500-fold higher than in mouse organs. CONCLUSION: Expression of alpha-gal epitopes in pig organs is many fold higher than in mouse organs. The abundance of these epitopes in pigs raises the question of whether pigs can properly develop without expression of alpha-gal epitopes.
BACKGROUND: Expression of the alpha-gal epitope in mice can be completely eliminated by disruption of the alpha1,3 galactosyltransferase gene. As an initial step for assessing the feasibility of this approach in the pig, it was of interest to compare the expression of alpha-gal epitopes in pig and mouse organs. METHODS: Membranes from pig and mouse organ homogenates were analyzed for alpha-gal epitope expression by Western blots, enzyme-linked immunosorbent assay (ELISA), immunostaining of tissues, and ELISA inhibition assay. RESULTS: Immunostaining of Western blots with human anti-Gal detected alpha-gal epitopes on glycoproteins from pig organs but not on glycoproteins from the corresponding mouse organs. ELISA with membrane homogenates and immunostaining of tissue sections demonstrated a much higher binding of human anti-Gal to alpha-gal epitopes on pig membranes than on mouse membranes. ELISA inhibition assay with monoclonal anti-Gal indicated that alpha-gal epitope expression in pig organs is up to 500-fold higher than in mouse organs. CONCLUSION: Expression of alpha-gal epitopes in pig organs is many fold higher than in mouse organs. The abundance of these epitopes in pigs raises the question of whether pigs can properly develop without expression of alpha-gal epitopes.
Authors: Carol J Phelps; Chihiro Koike; Todd D Vaught; Jeremy Boone; Kevin D Wells; Shu-Hung Chen; Suyapa Ball; Susan M Specht; Irina A Polejaeva; Jeff A Monahan; Pete M Jobst; Sugandha B Sharma; Ashley E Lamborn; Amy S Garst; Marilyn Moore; Anthony J Demetris; William A Rudert; Rita Bottino; Suzanne Bertera; Massimo Trucco; Thomas E Starzl; Yifan Dai; David L Ayares Journal: Science Date: 2002-12-19 Impact factor: 47.728
Authors: Donna Kolber-Simonds; Liangxue Lai; Steven R Watt; Maria Denaro; Scott Arn; Monica L Augenstein; Jeffery Betthauser; David B Carter; Julia L Greenstein; Yanhong Hao; Gi-Sun Im; Zhonghua Liu; Greg D Mell; Clifton N Murphy; Kwang-Wook Park; August Rieke; David J J Ryan; David H Sachs; Erik J Forsberg; Randall S Prather; Robert J Hawley Journal: Proc Natl Acad Sci U S A Date: 2004-05-03 Impact factor: 11.205