Abstract
The major obstacle to successful discordant xenotransplantation is the phenomenon of hyperacute rejection (HAR). In the pig-to-primate discordant transplant setting, HAR results from the deposition of high-titre anti-α-galactosyl antibodies and complement activation leading to endothelial cell destruction and rapid graft failure. To overcome HAR, we developed an enzymatic carbohydrate remodelling strategy designed to replace expression of the Galα-1,3-Gal xenoepitope on the surface of porcine cells with the non-antigenic universal donor human blood group O antigen, the α-1,2-fucosyl lactosamine moiety (H-epitope). Xenogenic cells expressing the human α-1,2-fucosyltransferase expressed high levels of the H-epitope and significantly reduced Galα-1,3-Cal expression. As a result, these cells were shown to be resistant to human natural antibody binding and complement-mediated cytolysis.
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References
Auchincloss, H. Xenogeneic transplantation: A review. Transplantation 46, 1–20 (1988).
Auchincloss, H. Xenografting: A review. Transplant. Rev. 4, 14–27 (1990).
Platt, J.L. et al. Transplantation of discordant xenografts: A review of progress. Immun. Today 11, 450–456 (1990).
Platt, J.L. & Bach, F.H. Discordant xenografting: challenges and controversies. Curr. Opin. Immun. 3, 735–739 (1991).
Bogman, M.J.J.T., Berden, J.H.M., Hagemann, F.H.M., Maass, C.N. & Keone, R.A.P. Patterns of vascular damage in the antibody-mediated rejection of skin xenografts in the mouse. Am. J. Path. 100, 727–738 (1980).
Bogman, M.J.J.T., Berden, J.H.M., Cornelissen, I.M.H.A., Maass, C.N. & Keone, R.A.P. The role of complement in the induction of acute antibody-mediated vasculitis of rat skin grafts in the mouse. Am. J. Path. 109, 97–106 (1982).
Dalmasso, A.P. et al. Mechanism of complement activation in the hyperacute rejection of porcine organs in primate recipients. Am. J. Path. 140, 1157–1166 (1992).
Leventhal, J.R. et al. Complement depletion prolongs discordant cardiac xenograft survival in rodents and non-human primates. Transplantn Proc. 25, 398–399 (1993).
Pruitt, S. et al. The effect of soluble complement receptor type 1 on hyperacute rejection of porcine xenografts. Transplantation 57, 363–370 (1994).
Leventhal, J.R. et al. Removal of baboon and human antiporcine IgG and IgM natural antibodies by immunoabsorption. Transplantation 59, 294–300 (1995).
Brewer, R.J. et al. Depletion of preformed natural antibody in primates for discordant xenotransplantation by continuous donor organ plasma perfusion. Transplantn Proc. 25, 385–386 (1993).
McCurry, K.R. et al. Human complement regulatory proteins protect swine-to-primate cardiac xenografts from humoral injury. Nature Med. 1, 423–427 (1995).
Fodor, W.L. et al. Expression of a functional human complement inhibitor in a transgenic pig as a model for the prevention of xenogeneic hyperacute organ rejection. Proc. natn. Acad. Set. U.S.A. 91, 11153–11157 (1994).
Rosengard, A.M. et al. Tissue expression of the human complement inhibitor, decay accelerating factor, in transgenic pigs. Transplantation 59, 1325–1333 (1995).
Sandrin, M.S., Vaughan, H.A., Dabkowski, P.L. & McKenzie, I.F.C. IgM antibodies in human serum reacts predominantly with Gal (al, 3)Gal epitopes. Proc. natn. Acad. Sci. U.S.A. 90, 11391–11395 (1993).
Sandrin, M.S., Vaughan, H.A. & McKenzie, I.F.C. Identification of Gal (αl,3)Gal as the major epitope for pig-to-human vascularised xenografts. Transplantn Rev. 8, 134–149 (1994).
Sandrin, M.S. & McKenzie, I.F.C. Gal (αl, 3)Gal, the major xenoantigen (s) recognised in pigs by human natural antibodies. Immunol. Rev. 141, 169–190 (1994).
Cooper, D.K.C. et al. Identification of a-galactosyl and other carbohydrate epitopes that are bound by human anti-pig antibodies: Relevance to discordant xenografting in man. Transplantn Immun. 1, 198–205 (1993).
Cooper, D.K.C., Koren, E. & Oriol, R. Oligosaccharides and discordant xenotransplantation. Immunol. Rev. 141, 31–58 (1994).
Good, A.H. et al. Identification of carbohydrate structures that bind antiporcine antibodies: Implications for discordant xenografting in humans. Tranplantn Proc. 24, 559–562 (1992).
Galili, U., Clark, M.R., Shohet, S.B., Buehler, J. & Macher, B.A. Evolutionary relationship between the natural anti-Gal antibody and the Galal-3Gal epitope in primates. Proc. natn. Acad. Sci. U.S.A. 84, 1369–1373 (1987).
Galili, U., Shohet, S.B., Korbin, E., Stults, C.L.M. & Macher, B.A. Man, apes and Old World monkeys differ from other mammals in the expression of the a-galactosyl epitopes on nucleated cells. J. biol. Chem. 263, 17755–17762 (1988).
Larsen, R.D. et al. Isolation of a cDNA encoding a murine UDPgalactose:β-D-galctosyl-1, 4-N-acetyl-glucosaminide-l,3-galactosyltransferase: Expression cloning by gene transfer. Proc. natn. Acad. Sci. U.S.A. 86, 8227–8231 (1989).
Joziasse, D.H., Shaper, N.L., Kim, D., Van den Eijnden, D.H. & Shaper, J.H. Murine α1,3 galactosyltransferase a single gene locus specifies four isoforms of the enzyme by alternative splicing. J. biol. Chem. 267, 5534–5541 (1992).
Joziasse, D.H., Shaper, J.H., Van den Eijnden, D.H., Van Tunen, A.J. & Shaper, N.L. Bovine α1,3 galactosyltiansferase: Isolation and characterization of a cDNA clone. Identification of homologous sequences in human genomic DNA. J. biol. Chem. 264, 14290–14297 (1989).
Sandrin, M.S., Dabkowski, P.L., Henning, M.M., Mouhtouris, E. & McKenzie, I.F.C. Characterization of cDNA clones for porcine α1,3 galactosyltransferase: The enzyme generating the Gal (α1,3)Gal epitope. Xenotransplantation 1, 81–88 (1994).
Joziasse, D.H., Shaper, J.H., Jabs, E.W. & Sharper, N.L. Characterization of an al, 3-galactosyltransferase homologue on human chromosome 12 that is organized as a processed pseudogene. J. biol. Chem. 266, 6991–6998 (1991).
Larsen, R.D., Riverra-Marrero, C.A., Ernst, L.K., Cummings, R.D. & Lowe, J.B. Frameshift and non sense mutations in a human genomic sequence homologous to a murine UDP-Gal:β-D-Gal 1,4-D-GlcNAcα1,3-galactosyl-transferase cDNA. J. biol. Chem. 265, 7055–7061 (1990).
Larsen, R.D., Ernst, L.K., Nair, R.P. & Lowe, J.B. Molecular cloning, sequence, and expression of a human GDP-L-fucose: β-D-galactoside 2-L-fucosyltransferase cDNA that can form the H blood group antigen. Proc. natn. Acad. Sci. U.S.A. 87, 6674–6678 (1990).
Fischel, R.J. et al. Plasma exchange, organ perfusion, and immunosuppression reduce “natural” antibody levels as measured by binding to xenogeneic endothelial cells and prolong discordant xenograft survival. Transplantn Proc. 24, 574–575 (1992).
Cooper, D.K.C. et al. Specific intravenous carbohydrate therapy: A new approach to the inhibition of antibody-mediated rejection following AB-incompatible allografting and discordant xenografting. Transplantn Proc. 25, 377–378 (1993).
Oriol, R., Ye, Y., Koren, E. & Cooper, D.K.C. Carbohydrate antigens of pig tissues reacting with human natural antibodies as potential targets for hyperacute vascular rejection in pig-to-man organ xenotransplantation. Transplantation 56, 1433–1442 (1993).
Kornfeld, R. & Kornfeld, S. Assembly of asparagine-linked Oligosaccharides. Annu. Rev. Biochem. 54, 631–664 (1985).
Schachter, H. Enzymes associated with glycosylation. Curr. Opin. struct. Biol. 1, 755–765 (1991).
Shaper, J.H. & Shaper, N.L. Enzymes associated with glycosylation. Curr. Opin. Struct. Biol. 2, 701–709 (1992).
Ausubel, F.M. et al. Current Protocols in Molecular Biology (Wiley-Interscience, New York, 1994).
Vaughan, H.A., Henning, M.M., Purcell, D.F.J., McKenzie, I.F.C. & Sandrin, M.S. Isolation of cDNA clones for CD48. Immunogenetics 33, 113–117 (1991).
Hayes, C.E. & Goldstein, I.J. An α-D-galactosyl-binding lectin from Bandeiraea simplicifolia seeds. J. biol. Chem. 249, 1904–1914 (1974).
Matsumoto, I. & Osawa, T. 1969. Purification and characterization of an anti-H (O) phytohemagglutinin of Ulex europeus. Biochim. biophys. Acta 194, 180–189 (1969).
Sandrin, M.S., Vaughan, H.A., McKenzie, I.F.C., Tait, B.D. & Parish, C.R. The human la system: Definition and characterization by xenogeneic antisera. Immunogenetics 8, 185–200 (1979).
Rajan, V.R., Larsen, R.D., Ajmera, S., Ernst, L.K. & Lowe, J.B. A cloned human DNA restriction fragment determines expression of a GDP-L, -fucose:β-D-galactoside 2 α-L-fucosyltransferase in transfected cells: Evidence for isolation and transfer of the human H blood group locus. J. biol. Chem. 264, 11158–11167 (1989).
Van der Eijnden, D.H., Blanken, W.M., Winterwerp, H. & Schiphorst, W.E.C.M. Identification and characterization of an UDP-Gal: N-acetyllactosaminide α-1, 3-D-galactosyltransferase in calf thymus. Eur. J. Biochem. 134, 523–530 (1983).
Brew, K., Vanaman, T.C. & Hill, R.L. The role of alpha-lactalbumin and the A protein in lactose synthetase: A unique mechanism for the control of a biological reaction. Proc. natn. Acad. Sci. U.S.A. 59, 491–497 (1968).
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Sandrin, M., Fodor, W., Mouthtouris, E. et al. Enzymatic remodelling of the carbohydrate surface of a xenogenic cell substantially reduces human antibody binding and complement-mediated cytolysis. Nat Med 1, 1261–1267 (1995). https://doi.org/10.1038/nm1295-1261
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DOI: https://doi.org/10.1038/nm1295-1261
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