The target antigens of naturally occurring human anti-beta-galactose IgG are cryptic on porcine aortic endothelial cells.

The identification of the xeno-antigens/xeno-antibodies combinations involved in pig-to-human xenograft rejection is an essential step for understanding this process and for the development of procedures to prevent it. Although it is widely accepted that the terminal disaccharide Galalpha1,3Gal-R is by far the major epitope recognized by human natural antibodies reactive with pig tissues, there is also evidence that other carbohydrate epitopes might be important in xenograft rejection. In an attempt to further improve our knowledge of the repertoire of human natural antibodies with anti-pig specificity we sought to determine whether naturally occurring human anti-beta-galactose IgG could interact with porcine aortic endothelial cells (PAEC). Histochemical analysis of porcine aorta sections revealed that the carbohydrate structures recognized by the anti-beta-galactose IgG are present on endothelial cells but in a cryptic form that can be unmasked by sialidase treatment. These structures were also found to be cryptic in cultured PAEC. In addition we demonstrated that PAEC may adsorb fetal calf serum (FCS) glycoproteins when cultured in FCS-supplemented medium, a process susceptible to generating artifactual observations in carbohydrate antigens analysis. In conclusion, despite their abundance, human anti-beta-galactose IgG do not represent a primary concern in pig-to-human xenotransplantation as the carbohydrate structures to which they bind are normally masked by sialic acid residues on porcine endothelial cells. However, whether these cryptic epitopes might be exposed on endothelial cells from genetically engineered animals should be further investigated because, if so, additional approaches will be needed to suppress their interaction with human anti-beta-galactose IgG.

Bacterial resistance of refrigerated and cryopreserved aortic allografts in an experimental virulent infection model.

PURPOSE: The bacterial resistance of refrigerated and cryopreserved aortic allografts in a highly virulent infection in a dog model was studied. METHODS: The infrarenal aorta of 12 dogs was replaced with either a cryopreserved aortic allograft (group I, n = 6) or a refrigerated aortic allograft (group II, n = 6) in infected sites. Allografts were harvested from dogs and stored for 1 week, either by cryopreservation (-140 degrees C) or refrigerated method (4 degrees C), in a preservation medium. At the time of implantation, induction of infection was achieved with an infected piece of knitted Dacron placed just beneath the allograft. The Dacron was contaminated in vitro by soaking it in a solution with Staphylococcus aureus PR209. All 12 dogs received no adjunct antibiotic or antithrombotic therapy. Four weeks after implantation, the animals were killed to recover the grafts for bacteriological and histological analyses. Bacterial results were expressed as colony-forming units (CFU)/cm2 of graft material. RESULTS: In group I, only one allograft grew bacteria at 2. 16 x 10(6 )CFU/cm2, with a blood culture positive for S aureus. In group II, one dog died at 3 weeks from a false septic aneurysm rupture, all the allografts were infected (P <.05) with a mean bacterial count of 9.41 +/- 6.8 x 10(4) CFU/cm2, and three blood cultures were positive for S aureus. The patency of the grafts was analyzed at the time of recovery. Three laminar thrombi without occlusion were present in group I; none were present in group II. A better preserved endothelium in group I was revealed by means of histologic analysis staining with factor VIII antibody before implantation. After 4 weeks of implantation in the infected site, infected allografts presented polynuclear infiltrates in the media with a high degree of inflammatory reaction, and endothelial recovery was more significant in group I, with numerous young plump cells. CONCLUSION: This study demonstrates that cryopreserved allografts implanted in infected sites in a dog model can produce greater bacterial resistance.