BACKGROUND: The expression of human complement regulators has been proved as an effective strategy to overcome hyperacute rejection in discordant xenogeneic organ transplantation. In this study, we tested the hypotheses that expression of triple transgenes for human complement regulators and provide more effective protection to the transplanted pig tissues. METHODS: Pigs transgenic for human complement regulatory proteins, human CD59 (hCD59) and human membrane cofactor protein (hMCP), have been generated using large genomic constructs. Heterozygous human decay-accelerating factor (hDAF) transgenic pigs, from a previously established line, were bred with hCD59 or hCD59 plus hMCP pigs to produce animals that expressed both hCD59 and hDAF, or expressed triple transgenes hCD59, hDAF and hMCP. RESULTS: All three transgenes were widely expressed in most of the tissues analyzed, but the expression of hMCP was at low levels. In cytotoxicity assays on porcine peripheral blood mononuclear cells, the expression of a single transgenic protein, hCD59, or hCD59 in combination with hMCP provided similar protection against human complement-mediated damage as the single expression of hDAF. However, the expression of triple transgenic proteins or double hCD59 and hDAF transgenic proteins provided greater protection than either hCD59 or hDAF alone. CONCLUSIONS: Thus, pigs transgenic for multiple transgenes provide a greater degree of human complement regulation and hence might be more suitable for xenotransplantation.
BACKGROUND: The expression of human complement regulators has been proved as an effective strategy to overcome hyperacute rejection in discordant xenogeneic organ transplantation. In this study, we tested the hypotheses that expression of triple transgenes for human complement regulators and provide more effective protection to the transplanted pig tissues. METHODS:Pigs transgenic for human complement regulatory proteins, humanCD59 (hCD59) and humanmembrane cofactor protein (hMCP), have been generated using large genomic constructs. Heterozygous human decay-accelerating factor (hDAF) transgenic pigs, from a previously established line, were bred with hCD59 or hCD59 plus hMCPpigs to produce animals that expressed both hCD59 and hDAF, or expressed triple transgenes hCD59, hDAF and hMCP. RESULTS: All three transgenes were widely expressed in most of the tissues analyzed, but the expression of hMCP was at low levels. In cytotoxicity assays on porcine peripheral blood mononuclear cells, the expression of a single transgenic protein, hCD59, or hCD59 in combination with hMCP provided similar protection against human complement-mediated damage as the single expression of hDAF. However, the expression of triple transgenic proteins or double hCD59 and hDAF transgenic proteins provided greater protection than either hCD59 or hDAF alone. CONCLUSIONS: Thus, pigs transgenic for multiple transgenes provide a greater degree of human complement regulation and hence might be more suitable for xenotransplantation.
Authors: K P Samy; R P Davis; Q Gao; B M Martin; M Song; J Cano; A B Farris; A McDonald; E K Gall; C R Dove; F V Leopardi; T How; K D Williams; G R Devi; B H Collins; A D Kirk Journal: Am J Transplant Date: 2017-12-28 Impact factor: 8.086
Authors: Yonglun Luo; Lin Lin; Lars Bolund; Thomas G Jensen; Charlotte Brandt Sørensen Journal: J Inherit Metab Dis Date: 2012-03-28 Impact factor: 4.982
Authors: B M Martin; K P Samy; M C Lowe; P W Thompson; J Cano; A B Farris; M Song; C R Dove; F V Leopardi; E A Strobert; J B Jenkins; B H Collins; C P Larsen; A D Kirk Journal: Am J Transplant Date: 2015-02-19 Impact factor: 8.086
Authors: Robert J Plenter; Todd J Grazia; An N Doan; Ronald G Gill; Biagio A Pietra Journal: J Heart Lung Transplant Date: 2012-07-11 Impact factor: 10.247