title: Morphology of hDAF (CD55) transgenic pig kidneys following ex-vivo hemoperfusion with human blood.
abstract: Discordant xenotransplantation of pig kidneys into man may be possible in the future using transgenic organs which regulate complement activity. It was the aim of this experimental study to characterize morphologic alterations of organs transgenic for human decay accelerating factor (hDAF/CD55) perfused with human blood since no data on function of these organs after exposure to human blood are available. An ex-vivo system was developed that allows computer driven pressure-controlled perfusion of kidneys including a separate cartridge oxygenator circuit. Following cold ischemia time of 1-4 hr, 8 kidneys from heterozygote transgenic animals (TG) and 9 control kidneys (C) were perfused with 500 ml freshly drawn heparinized human blood at physiological conditions. A histologic grading system from 0 to +4 was used to describe the histologic findings. Using a mouse antihuman DAF moAB, hDAF was stained on all TG kidneys both on glomerular capillary (4+) and vascular endothelium (2+), but there was no detectable hDAF-expression on controls. No difference in xenoantibody deposition on vascular endothelium was seen between both groups. There was comparable staining for complement fraction C4 in both groups, but significant reduction of C3 and C9 staining on glomerular and vascular endothelium in TG. P-selectin was expressed on a higher level in C (+4) compared with TG (+2). Neutrophil extravasation [NP-57 elastase] was higher in C (80.2 vs. 32.2 C vs. TG [values as n/high power field]). Tubular epithelial cell swelling and mild necrosis was paralleled by glomerular hemorrhage and platelet microthrombus formation in both groups as seen in transmission electron microscopy. The observed results allow the conclusion that hDAF expression on transgenic pig kidneys was sufficient to inhibit complement activation beyond C3 during xenoperfusion with human blood despite xenoantibody deposition.
authors.affiliation: Institute for Surgical Research and Department of Pathology, Ludwig-Maximilian University of Munich, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Storck
authors.forename: D
authors.initials: D
authors.lastname: Abendroth
authors.forename: R
authors.initials: R
authors.lastname: Prestel
authors.forename: G
authors.initials: G
authors.lastname: Pino-Chavez
authors.forename: J
authors.initials: J
authors.lastname: Müller-Höker
authors.forename: D J
authors.initials: DJ
authors.lastname: White
authors.forename: C
authors.initials: C
authors.lastname: Hammer
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D000911:Antibodies, Monoclonal; D015703:Antigens, CD / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics; D018958:CD55 Antigens / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*; D016207:Cytokines / Q000097:blood*; D004797:Enzyme-Linked Immunosorbent Assay; D006464:Hemoperfusion / Q000295:instrumentation / Q000379:methods; D006579:Heterozygote; D006801:Humans; D007150:Immunohistochemistry; D007375:Interleukin-1 / Q000097:blood; D016753:Interleukin-10 / Q000097:blood; D015850:Interleukin-6 / Q000097:blood; D007668:Kidney / Q000276:immunology* / Q000473:pathology / Q000648:ultrastructure; D007687:Kidney Tubules, Proximal / Q000473:pathology / Q000648:ultrastructure; D051379:Mice; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis; D012680:Sensitivity and Specificity; D013552:Swine; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis
chemical_list: D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D018958:CD55 Antigens; D016207:Cytokines; D007375:Interleukin-1; D015850:Interleukin-6; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha; D016753:Interleukin-10