Adenovirus-mediated gene transfer of triple human complement regulating proteins (DAF, MCP and CD59) in the xenogeneic porcine-to-human transplantation model. Part II: xenogeneic perfusion of the porcine liver in vivo.
In this study, the adenovirus-mediated gene transfer of triple human complement regulating proteins was investigated in xenogeneic pig liver perfusion. The porcine liver was perfused in situ at 4 degrees C under a pump-driven veno-venous shunt of the portal vein and inferior vena cava, with 5 to 15x10(11) plaque-forming units (pfu) of adenovirus vector (group 1: AxCALacZ; 2: AxCACD59; 3: AxCACD59 + AxCADAF; 4: AxCACD59 + AxCADAF + AxCAMCP) for 1 h (for each, n=3). The livers were harvested 24 h after gene transfer and then were reperfused ex-vivo with fresh human blood for 2 h. In immunohistochemical staining, each complement regulating protein (CRP) showed a distribution similar to that of the LacZ expression. The C3 levels in the perfusate were also maintained at higher levels in group 4 from 60 to 120 min after reperfusion (C3: 85% to 95% of the initial level) than in groups 1 to 3 (C3: 80% to 90% of the initial level) from 60 to 120 min after reperfusion. The complement deposition on the porcine liver [C3, membrane attack component (MAC)] decreased significantly more in group 4 than in groups 1 to 3. In conclusion, the adenovirus-mediated multiple gene transfer of human CRPs (hCRPs) was found to effectively suppress the complement activation in xenogeneic pig liver perfusion.
Shiraishi
Masayuki
M
First Department of Surgery, University of the Ryukyus, School of Medicine, Uehara 207, Nishihara-cho, Okinawa 903-0215, Japan. masa@med.u-ryukyu.ac.jp
Oshiro
Takashi
T
Nozato
Eiji
E
Nagahama
Masayoshi
M
Taira
Kaoru
K
Nomura
Hironori
H
Sugawa
Hideki
H
Muto
Yoshihiro
Y
D000256:Adenoviridae; D000818:Animals; D015703:Antigens, CD / Q000235:genetics* / Q000276:immunology; D018958:CD55 Antigens / Q000235:genetics* / Q000276:immunology; D018957:CD59 Antigens / Q000235:genetics* / Q000276:immunology; D003165:Complement System Proteins / Q000276:immunology / Q000502:physiology*; D005697:Galactosides / Q000032:analysis; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D006801:Humans; D008099:Liver / Q000276:immunology / Q000502:physiology*; D051916:Membrane Cofactor Protein; D008562:Membrane Glycoproteins / Q000235:genetics* / Q000276:immunology; D018448:Models, Immunological; D010477:Perfusion; D013552:Swine; D014183:Transplantation, Heterologous / Q000276:immunology*; D001616:beta-Galactosidase / Q000235:genetics
D015703:Antigens, CD; C497857:CD46 protein, human; D018958:CD55 Antigens; D018957:CD59 Antigens; D005697:Galactosides; D051916:Membrane Cofactor Protein; D008562:Membrane Glycoproteins; D003165:Complement System Proteins; D001616:beta-Galactosidase