An in vitro model of pig liver xenotransplantation-pig complement is associated with reduced lysis of wild-type and genetically modified pig cells

Background: After pig liver transplantation in humans, the graft will produce pig complement (C). We investigated in vitro the lysis of wild-type (WT), alpha 1,3-galactosyltransferase gene-knockout (GTKO), and CD46 transgenic (CD46) pig peripheral blood mononuclear cells (PBMC) caused by human anti-pig antibodies (Abs) + pig C. Methods: Human serum IgM/IgG binding to WT and GTKO PBMC was determined by flow cytometry, and lysis of pig PBMC by a C-dependent cytotoxicity assay using (i) human serum (human Abs + C), (ii) GTKO pig serum (anti-Gal Abs + pig C), (iii) heat-inactivated human serum (human Abs) + rabbit C, or (iv) human Abs + pig C (serum). Results: Binding of human IgM and IgG to GTKO PBMC was less than to WT PBMC (P < 0.05). In the presence of human Abs, lysis of WT and GTKO PBMC by rabbit C was 87 and 13%, respectively (WT vs. GTKO, P < 0.01), but was only 37 and 0.4% in the presence of pig C (WT vs. GTKO, P < 0.05). Human/rabbit C-induced lysis was greater than pig C-induced lysis for both WT and GTKO PBMC. CD46 pig PBMC reduced rabbit/human C- and pig C-mediated lysis (P < 0.05). Conclusions: Pig livers, particularly from GTKO and CD46 pigs, are likely to have an immunologic advantage over other organs after transplantation into humans. In the absence of pig antibodies directed to human tissues, pig complement is unlikely to cause problems after liver xenotransplantation, especially if GTKO/CD46 pigs are used as the source of the livers.