Abstract
Hyperacute rejection (HAR) occurring after transplantation within phylogenetically distant species is a severe reaction triggered by preexisting xenoreactive antibodies and complement activation, leading to the destruction of the donor organ. Expression of human complement inhibitors in transgenic pig organs prolongs the survival of xenograft in experimental models. Moreover, the extent of protection from hyperacute rejection is dependent on the level and site of expression of the transgenic molecules and, probably, on the combination of different molecules. In this regard a small animal model to test the efficacy of expression vectors and different human molecules could be very advantageous. A murine model developed in our laboratory was characterized by measurement of several parameters characteristic of HAR in the livers of control and transgenic mice expressing transgenic human DAF (CD55) or MCP (CD46) at the end of 2 h of perfusion with human plasma and after 1 day. The parameters studied were heamatological values of hepatic functions (GOT and GPT), induction of pro-inflammatory molecules and histopathological evaluation. Cytokines (IL-1α, IL-1β, IL-6) induction and exposure of P-selectin on the endothelial cell surface, was only observed in control animals after 2 h of perfusion, as an early event. GOT and GPT values increase drammatically after 2 h perfusion and 1 day after the treatment according to the histopathological observation of liver damage. On the contrary, the livers of hDAF or hMCP transgenic mice, under the same treatment were significantly protected although the extent of this protection is dependent on the level of expression of transgenic human molecules.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bach FH, Robson SC, Winkler H, Ferran C, Stuhlmeier KM, Wrighton CJ and Hancock WW (1995) Barriers to xenotransplantation. Nat Med 1: 869–873.
Bach FH (1997) Genetic engineering as an approach to xenotransplantation. World J Surgery 21: 913–916.
Bach FH, Ferran C, Hechenleitner P, Mark W, Koyamada N, Miyatake T, Winkler H, Badrichani A, Candinas D and Hancock WW (1997a) Accomodation of vascularized xenografts: expression of “protective genes” by donor endothelial cells in a host Th2 cytokine environment. Nat Med 3: 196–204.
Bach FH, Ferran C, Soares M, Wrighton CJ, Anrather J, Winkler H, Robson SC and Hancock WW (1997b) Modification of vascular responses in xenotransplantation: inflammation and apoptosis. Nat Med 3: 944–948.
Badrichani AZ, Stroka DM, Bilbao G, Curiel DT, Bach FH and Ferran C (1999) Bcl-2 and Bcl-XL serve an anti-inflammatory function in endothelial cells through inhibition of NF-kappaB. J Clin Invest 103: 543–553.
Byrne GW, McCurry KR, Martin MJ, McClellan SM, Platt JL and Logan JS (1997) Transgenic pigs expressing human CD59 and decay accelerating factor produce an intrinsic barrier to complement-mediated damage. Transplantation 63: 149–155.
Carrington CA, Richards AC, van den Bogaerde J, Tucker AW and White DJ (1997) Complement activation, its consequences, and blockade by gene transfer. World J Surgery 21: 907–912.
Chomcyznski P and Sacchi N (1987) Single-step method of isolation of RNA by guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162: 156–159.
Coughlan AF, Berndt MC, Dunlop LC and Hancock WW (1993) In vivo studies of P-selectin and platelet activating factor during endotoxemia, accelerated allograft rejection, and discordant xenograft rejection. Transplant Proc 25: 2930–2931.
Cozzi E and White DJ (1995) The generation of transgenic pigs as potential organ donors for humans. Nat Med 1: 964–966.
Dalmasso AP, Vercellotti GM, Fischel RJ, Bolman RM, Bach FH and Platt JL (1992) Mechanism of complement activation in the hyperacute rejection of porcine organs transplanted into primate recipients. Am J Pathol 140: 1157–1166.
Diamond LE, McCurry KR, Oldham ER, Tone M, Waldmann H, Platt JL and Logan JS (1995) Human CD59 expressed in transgenic mouse hearts inhibits the activation of complement. Transplant Immunol 3: 305–312.
Fodor WL, Williams BL, Matis LA, Madri JA, Rollins SA, Knight JW, Velander W and Squinto SP (1994) Expression of functional human complement inhibitor in a transgenic pig as a model for the prevention of xenogeneic hyperacute organ rejection. Proc Natl Acad Sci USA 91: 11153–11157.
Hogan BL, Costantini F and Lacy E (1986) Manipulating the Mouse Embryo: A Laboratory Manual, Cold Spring Harbor Lab Press, Cold Spring Harbor, NY.
Hourcade D, Holers VM and Atkinson JP (1989) The regulators of complement activation (RCA) gene cluster. Adv Immunol 45: 381–416.
Kroshus TJ, Bolman RM III, Dalmasso AP, Rollins SA, Guilmette ER, Williams BL, Squinto SP and Fodor WL (1996) Expression of human CD59 in transgenic pig organs enhances organ survival in an ex vivo xenogeneic perfusion model. Transplantation 61: 1513–1521.
Lazzeri M, Mora M, Mulder LCF, Marsicano G, Marinucci G, Boschi M, Bruzzone P, Alfani D, Cortesini R and Rossini M (1998) Kidneys derived from mice transgenic for human com plement blockers are protected in an in vivo model of hyperacute rejection J Urol 159: 1364–1369.
Lublin DM and Atkinson JP (1989) Decay-accelerating factor and membrane cofactor protein. Curr Top Microbiol Immunol 153: 123–145.
McCurry KR, Kooyman DL, Alvarado CG, Cotterell AH, Martin MJ, Logan JS and Platt JL (1995) Human complement regulatory proteins protect swine-to-primate cardiac xenografts from humoral injury. Nat Med 1: 423–427.
McKenzie IF, Li YQ, Patton K, Thall AD and Sandrin MS (1998) A murine model of antibody-mediated hyperacute rejection by galactose-alpha(1,3)galactose antibodies in Gal o/o mice. Transplantation 66: 754–763.
Mora M, Mulder LCF, Lazzeri M, Boschi M, Ciccopiedi E, Melli CM, Bruzzone P, Alfani D, Cortesini R and Rossini M (1996) Protection from complement-mediated injury in livers and kidneys of transgenic mice expressing human complement regulators. Xenotransplantation 3: 63–68.
Mulder LCF, Mora M, Ciccopiedi E, Melli C, Nuti S, Marinucci G, Bruzzone P, Lazzeri M, Lorenzini R, Alfani D, Cortesini R and Rossini M (1995a) Mice transgenic for human CD46 and CD55 are protected from human complement attack. Transplant Proc 27: 333–335.
Mulder LCF, Ciccopiedi E, Mora M, Nuti S, Marinucci G, Lazzeri M, Melli C, Marchetti M, Bruzzone P, Alfani D, Cortesini R and Rossini M (1995b) Livers of mice transgenic for human CD46 are protected from human complement attack. Transgenics 1: 629–637.
Palmiter RD, Sandgren EP, Koeller M and Brinster RL (1993) Distal regulatory elements from the mouse metallothionein locus stimulate gene expression in transgenic mice. Mol Cell Biol 13: 5266–5275.
Pierson RN III, Pino-Chavez G, Young VK, Kaspar-Konig W, White DJ and Wallwork J (1997) Expression of human decay accelerating factor may protect pig lung from hyperacute rejection by human blood. J Heart Lung Transplant 16: 231–239.
Romanella M, Aminian A, Adam WR, Pearse MJ and d'Apice AJ (1997) Involvement of both the classical and alternate pathways of complement in and ex vivo model of xenograft rejection. Transplantation 63: 1021–1025.
Saadi S, Holzknecht RA, Patte CP, Stern DM and Platt JL (1995) Complement-mediated regulation of tissue factor activity in endothelium. J ExpMed 182: 1807–1814.
Samuelsson BE, Rydberg L, Breimer ME, Backer A, Gustavsson M, Holgersson J, Karlsson E, Uyterwaal AC, Cairns T and Welsh K (1994) Natural antibodies and human xenotransplantation. Immunol Rev 141: 151–168.
Sandrin MS and McKenzie IF (1994) Galα(1,3)Gal, the major xenoantigen(s) recognized in pigs by human natural antibodies. Immunol Rev 141: 169–190.
Selvan RS, Kapadia HB and Platt JL (1998) Complement-induced expression of chemokine genes in endothelium: regulation by IL-1-dependent and-indipendent mechanisms. J Immunol 161: 4388–4395.
Sharma A, Okabe J, Birch P, McClellan SB, Martin MJ, Platt JL and Logan JS (1996) Reduction in the level of Gal(α1,3)Gal in transgenic mice and pigs by the expression of an α (1,2)fucosyltransferase. Proc Nat Acad Sci USA 93: 7190–7195.
Soares MP, Lin Y, Anrather J, Csizmadia E, Takigami K, Sato K, Grey ST, Colvin RB, Choi AM, Poss KD and Bach FH (1998) Expression of heme oxygenase-1 can determine cardiac xenograft survival. Nat Med 4: 1073–1077.
Tran TH, Grey S, Anrather J, Steinhauslin F, Bach FH and Winkler H (1998) Regulated and endothelial cell-specific expression of Fas ligand: an in vitro model for a strategy aiming at inhibiting xenograft rejection. Transplantation 66: 1126–1131.
van Denderen BJ, Pearse MJ, Katerelos M, Nottle MB, Du ZT, Aminian A, Adam WR, Shenoy-Scaria A, Lublin DM, Shinkel TA and d'Apice AJ (1996) Expression of functional decay-accelerating factor (CD55) in transgenic mice protects against human complement-mediated attack. Transplantation 61: 582–588.
van Denderen BJ, Salvaris E, Romanella M, Aminian A, Katerelos M, Tange MJ, Pearse MJ and d'Apice AJ (1997) Combination of decay-accelerating factor expression and alpha 1,3-galactosyltransferase knockout affords added protection from human complement-mediated injury. Transplantation 64: 882–886.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Mora, M., Lazzeri, M., Marsicano, G. et al. An in vivo model of hyperacute rejection: characterization and evaluation of the effect of transgenic human complement inhibitors. Transgenic Res 9, 205–213 (2000). https://doi.org/10.1023/A:1008928713058
Issue Date:
DOI: https://doi.org/10.1023/A:1008928713058