Hyperacute rejection of hDAF-transgenic pig organ xenografts in cynomolgus monkeys: influence of pre-existing anti-pig antibodies and prevention by the alpha GAL glycoconjugate GAS914.
BACKGROUND
Our introductory pig-to-cynomolgus monkey heart or kidney transplantation using organs from pigs transgenic for human decay-accelerating factor (hDAF), showed a high incidence of hyperacute rejection (HAR), which was ascribed to extraordinary high levels of anti-pig antibodies. We evaluated the efficacy of GAS914, a Gal alpha 1-3Gal trisaccharide linked to a poly-l-lysine backbone, in inhibition of HAR.


METHODS
hDAF transgenic heterotopic heart (n = 15) or life-supporting kidney (n = 8) transplantation included induction with cyclophosphamide or anti-thymocyte globulin, and maintenance with cyclosporine or tacrolimus, steroids and mycophenolate sodium/mofetil. Four doses of GAS914 were given before transplantation. Rejection was confirmed by graft histology, and anti-pig antibody levels were determined in various assays.


RESULTS
Four of six heart transplants without GAS914 treatment showed HAR. Nine subsequent transplants with GAS914 pre-treatment, did not show HAR (chi-square, P < 0.05). Two of four kidney transplants without GAS914 treatment ended with HAR. Four subsequent transplants with GAS914 did not show HAR. Animals with HAR showed extremely high antibody levels. Samples just before transplantation showed significantly higher antibody levels in recipients presenting with HAR. In all assays antibody levels were significantly lowered by GAS914 pre-treatment.


CONCLUSIONS
HAR of hDAF solid organs could be ascribed to high levels of anti-pig antibodies. It is hypothesized that the hDAF transgene shows a threshold in efficacy, above which an overwhelming attack by antibodies and complement activation cannot be modulated to prevent HAR. HAR does not occur when animals with lower levels are used, or when antibodies are effectively depleted from the circulation by GAS914 treatment.
Transplantation Immunology, Department of Cardiothoracic Surgery, Stanford University School of Medicine, Stanford, CA 94305-5407, USA.
Tuan T
TT
Lam
Bernard
B
Hausen
Katrin
K
Boeke-Purkis
Ricardo
R
Paniagua
Macy
M
Lau
Laurie
L
Hook
Gerry
G
Berry
John
J
Higgins
Rudolf O
RO
Duthaler
Andreas G
AG
Katopodis
Robert
R
Robbins
Bruce
B
Reitz
Dominique
D
Borie
Henk-Jan
HJ
Schuurman
Randall E
RE
Morris
D000208:Acute Disease; D000818:Animals; D030801:Animals, Genetically Modified; D000910:Antibodies, Heterophile / Q000097:blood / Q000276:immunology*; D006084:Graft Rejection / Q000453:epidemiology / Q000276:immunology*; D006085:Graft Survival / Q000276:immunology; D016027:Heart Transplantation / Q000276:immunology*; D015994:Incidence; D016030:Kidney Transplantation / Q000276:immunology; D008252:Macaca fascicularis; D008297:Male; D034421:Sus scrofa; D014183:Transplantation, Heterologous / Q000276:immunology*; D014312:Trisaccharides / Q000276:immunology / Q000494:pharmacology*
D000910:Antibodies, Heterophile; D014312:Trisaccharides; C076569:alpha-galactosyl epitope