title: The effect of soluble complement receptor type 1 on acute humoral xenograft rejection in hDAF-transgenic pig-to-primate life-supporting kidney xenografts.
abstract: BACKGROUND
  In pig-to-nonhuman primate solid organ xenotransplantation using organs from donors transgenic for human decay-accelerating factor (hDAF), the main type of rejection is antibody-mediated (acute humoral xenograft rejection, AHXR). This occurs despite the complement-regulatory function of the transgene, neutralization of natural antibodies to Galalpha1-3Gal (Gal) using soluble glycoconjugates, and chronic immunosuppression. As complement components play a major role in graft destruction after antibody binding, we evaluated the efficacy of chronic complement inhibition by soluble complement receptor type 1 (TP10).
  
  
  METHODS
  Life-supporting hDAF-transgenic kidney transplantation was performed in cynomolgus monkeys, using cyclophosphamide induction, and maintenance immunosuppression with cyclosporin A, mycophenolate sodium, and tapering steroids. Rejection was treated with bolus steroid injections: if not successful animals were terminated. Three groups were studied: in group 1 (n=4) GAS914 (a soluble glycoconjugate comprising Gal on a poly-L-lysine backbone) was added before and after transplantation; group 2 (n=2) received GAS914 as in group 1 and in addition TP10 before and after transplantation; in group 3 (n=4) GAS914 was only given before transplantation and TP10 as in group 2. Monitoring included the regular assessment of anti-porcine antibodies, complement activity (soluble C5b-9), therapeutic drug monitoring, and graft histology.
  
  
  RESULTS
  Survival in group 1 was 6, 12, 31 and 37 days, respectively, and in all four cases graft histology showed AHXR. The two animals in groups 2 survived 3 and 15 days, respectively, and similarly showed AHXR in graft histology. In group 3 two animals showed AHXR (10 and 37 days survival, respectively), and two others did not show AHXR (20 and 32 days survival, respectively). The diagnosis AHXR included the deposition of complement activation products in the graft, which were present at lower intensity in animals treated with TP10. In all animals GAS914 effectively neutralized circulating anti-Gal antibody. Antibodies were detectable in the circulation of all animals using porcine erythrocytes in a hemolytic assay, although at lower levels than before transplantation. Soluble C5b-9 was not detectable in the circulation of animals receiving TP10, and circulating TP10 concentrations in these animals were in a presumed pharmacologically active range.
  
  
  CONCLUSIONS
  The inclusion of TP10 in the immunosuppressive protocol does not clearly lead to improved xenograft survival. Despite effective neutralization of anti-Gal antibodies and effective inhibition of systemic complement activity, AHXR was apparent in four of six animals under chronic TP10 treatment, including deposits of complement activation products in the graft. Apparently, effective systemic complement inhibition by TP10 in combination with local complement regulation by the hDAF transgene product does not necessarily result in effective inhibition of complement activation at locations in the xenograft upon binding of anti-porcine antibodies to the grafted endothelium.
authors.affiliation: Transplantation Immunology, Department of Cardiothoracic Surgery, Stanford University School of Medicine, Stanford, CA 94305-5407, USA.
authors.forename: Tuan T
authors.initials: TT
authors.lastname: Lam
authors.forename: Bernard
authors.initials: B
authors.lastname: Hausen
authors.forename: Laurie
authors.initials: L
authors.lastname: Hook
authors.forename: Macy
authors.initials: M
authors.lastname: Lau
authors.forename: John
authors.initials: J
authors.lastname: Higgins
authors.forename: Uwe
authors.initials: U
authors.lastname: Christians
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Jacobsen
authors.forename: Muhammad
authors.initials: M
authors.lastname: Baluom
authors.forename: Rudolf
authors.initials: R
authors.lastname: Duthaler
authors.forename: Andreas
authors.initials: A
authors.lastname: Katopodis
authors.forename: Gilda
authors.initials: G
authors.lastname: Chavez
authors.forename: Emanuele
authors.initials: E
authors.lastname: Cozzi
authors.forename: Richard
authors.initials: R
authors.lastname: Harrison
authors.forename: Henk-Jan
authors.initials: HJ
authors.lastname: Schuurman
authors.forename: Dominique
authors.initials: D
authors.lastname: Borie
authors.forename: Randall E
authors.initials: RE
authors.lastname: Morris
mesh_terms: D000208:Acute Disease; D000818:Animals; D030801:Animals, Genetically Modified; D018958:CD55 Antigens / Q000276:immunology*; D003169:Complement Inactivator Proteins / Q000494:pharmacology*; D015938:Complement Membrane Attack Complex / Q000378:metabolism; D016572:Cyclosporine / Q000494:pharmacology; D004359:Drug Therapy, Combination; D006084:Graft Rejection / Q000188:drug therapy* / Q000276:immunology* / Q000473:pathology; D006085:Graft Survival / Q000276:immunology; D007166:Immunosuppressive Agents / Q000494:pharmacology; D016030:Kidney Transplantation / Q000276:immunology*; D008252:Macaca fascicularis; D009173:Mycophenolic Acid / Q000494:pharmacology; D011951:Receptors, Complement; D013256:Steroids / Q000494:pharmacology; D013552:Swine; D014183:Transplantation, Heterologous / Q000276:immunology*
chemical_list: D018958:CD55 Antigens; D003169:Complement Inactivator Proteins; D015938:Complement Membrane Attack Complex; D007166:Immunosuppressive Agents; D011951:Receptors, Complement; D013256:Steroids; C082885:soluble complement inhibitor 1; D016572:Cyclosporine; D009173:Mycophenolic Acid