Porcine islet cells of Langerhans are destroyed by human complement and not by antibody-dependent cell-mediated mechanisms.

Abstract

In the near future, xenotransplantation of porcine islets of Langerhans might be an alternative in the treatment of patients with diabetes mellitus. However, xenotransplantation of islets of Langerhans in large animals has been shown to result in an exceedingly short graft survival, which suggests that a humoral immune response might play a major role in islet demise. This study was performed to assess binding human preformed antibodies to isolated porcine islet cells (PIC) and to determine the lysis of PIC using human sera in complement-mediated cytotoxicity (CMC) and antibody-dependent cell-mediated cytotoxicity (ADCC) assays. Ten Dutch Landrace pigs were used for the isolation of PIC. Sera from 30 healthy blood donors (1/10 diluted) were used in a 51Cr release assay to assess CMC. Heat-inactivated normal human sera and fresh sera from patients with agammaglobulinemia were used as controls. Binding of human IgM IgG, and IgA antibodies to PIC was tested in an ELISA using isotype-specific secondary monoclonal antibodies ADCC was tested in a 51Cr release assay using normal human sera and sera from newly diagnosed type I diabetics with peripheral blood mononuclear cells as effector cells and PIC as targets. It was found that PIC were recognized by human IgM and IgG preformed antibodies and that fresh human sera had strong CMC activity inducing a percentage-specific PIC lysis of 61 +/- 10% (mean +/- SD) within 60 min. Agammaglobulinemic sera killed 42 +/- 12% of PIC. No significant cytotoxic activity was found in ADCC assays using normal sera or sera from diabetic patients. These results show that all tested human sera lyse PIC via CMC, even in the absence of human antibodies, as concluded from the use of agammaglobulinemic sera. In pig-to-human transplantation, islets may be hyperacutely rejected by antibody-dependent and antibody-independent activation of complement and not by antibody-dependent cell-mediated mechanisms.