BACKGROUND Our goal was to improve islet transplantation as a therapy for patients with type I diabetes mellitus. Because human donor islets are scarce, we are studying islet xenografts in the diabetic NOD mouse model. We hypothesize that optimal xenoislet survival will be achieved by the combination of donor islet immunoisolation with recipient immunosuppression. We and others have studied adult and neonatal porcine islets as sources of tissue for microencapsulated islet xenografts, but we believe it is also advantageous to consider using islets from fish, which can be raised in large numbers relatively quickly and economically. Therefore, in this study, we have evaluated the function of microencapsulated xenogeneic piscine (tilapia) islets transplanted intraperitoneally (IP) in NOD mice in the presence of CD4(+) T-cell depletion and/or costimulatory blockade. METHODS Spontaneously diabetic NOD mice or streptozotocin (STZ)-diabetic NOD-SCID mice were transplanted IP with microencapsulated tilapia islets. Recipient immunosuppression included anti-CD4 mAb, CTLA4-Ig, anti-CD80 mAb, anti-CD86 mAb, or anti-CD154 mAb, alone or in combination. Graft function was evaluated by blood glucose (BG) levels, intravenous (IV) and oral glucose tolerance tests (GTTs), histologic and immunohistochemical analyses of grafts, and flow cytometric analysis of peritoneal cells. RESULTS Encapsulated tilapia islets normalized random BG levels for up to 210 days in NOD-SCID mice. In diabetic NOD mice, encapsulated tilapia islets were rejected on day 11 ± 4 with a peritoneal infiltrate of macrophages, eosinophils, B cells, occasional neutrophils, but few T cells. Immunohistochemical staining demonstrated the presence of murine IgG on tilapia islets within capsules of rejecting, non-immunosuppressed mice, as well as murine IgG-positive lymphocytes in the layer of host cells surrounding those capsules. These findings suggested that our barium (Ba)-gelled alginate capsules are permeable to IgG and that anti-piscine antibodies may be involved in the rejection of encapsulated tilapia islets in untreated mice. No single immunosuppressive agent prolonged encapsulated tilapia islet survival in NOD mice, but the combination of CTLA4-Ig plus anti-CD154 mAb extended tilapia islet graft survival until rejection at 119 ± 20 days and inhibited host cell recruitment to the peritoneal cavity. Triple treatment with CTLA4-Ig, anti-CD154 mAb, and anti-CD4 mAb allowed graft survival for 157 ± 35 days with little evidence of a host cellular reaction. IV and oral glucose tolerance tests (GTTs) of recipients with functioning xenografts demonstrated remarkably normal metabolic function. CONCLUSIONS We conclude that microencapsulated tilapia islets can survive long term with excellent metabolic control in diabetic mice given targeted immunosuppression, suggesting that cross-species physiological incompatibility may not compromise the applicability of this novel approach for future clinical applications. We predict that an improved microcapsule that prevents the entrance of IgG will enhance tilapia islet survival in this model, possibly allowing the application of this technique with limited or no immunosuppression.