Combination therapy with cyclosporine and interleukin-4 or interleukin-10 prolongs survival of synergeneic pancreatic islet grafts in nonobese diabetic mice: islet graft survival does not correlate with mRNA levels of type 1 or type 2 cytokines, or transforming growth factor-beta in the islet grafts.
BACKGROUND
The recurrent autoimmune response to syngeneic pancreatic islet grafts transplanted into nonobese diabetic (NOD) mice is cell-mediated and relatively resistant to cyclosporine (CsA) therapy. Therefore, we asked whether interleukin (IL)-4 and IL-10, cytokines that inhibit cell-mediated immunity, might improve the therapeutic effect of CsA.


METHODS
We compared the survival of syngeneic islet grafts transplanted into diabetic NOD mice treated with IL-4, IL-10, and CsA, administered as single agents and in combinations. Additionally, we measured mRNA levels of type 1 cytokines (interferon-gamma [IFN-gamma], IL-2, and IL-12), type 2 cytokines (IL-4 and IL-10), and transforming growth factor-beta (TGF-beta) to determine whether graft rejection or survival might correlate with expression of these cytokines in the grafts.


RESULTS
CsA (20 mg/kg/day) significantly prolonged islet graft survival (median: 20 days vs. 10 days for vehicle-treated mice). Neither IL-4 (2.5 microg, twice daily), nor IL-10 (10 microg, twice daily) significantly prolonged islet graft survival. By contrast, combination therapy with CsA and IL-10 significantly prolonged islet graft survival (median: 34 days) compared with vehicle-treated mice (median: 10 days), and combination therapy with CsA and IL-4 significantly prolonged islet graft survival (median: 59 days) compared with both vehicle-treated mice (median: 10 days) and mice treated with CsA alone (median: 20 days). Islet grafts from normoglycemic mice treated with CsA plus IL-10, and with CsA plus IL-4, were surrounded but not infiltrated by mononuclear leukocytes and beta cells were intact, whereas islet grafts from mice treated with vehicle, IL-4, IL-10, and CsA (as single agents) were infiltrated by mononuclear leukocytes and fewer beta cells were detected. Polymerase chain reaction analysis of cytokine mRNA expression in islet grafts at 8-12 days after transplantation revealed that CsA decreased mRNA levels of type 1 cytokines (IFN-gamma and IL-12p40), whereas CsA plus IL-10 did not, and CsA plus IL-4 increased mRNA levels of IFN-gamma, IL-12p40, and TGF-beta.


CONCLUSIONS
These results demonstrate that IL-4, and to a lesser extent IL-10, improves the ability of CsA to prevent autoimmune destruction of beta cells in syngeneic islets transplanted into diabetic NOD mice; however, there is no simple correlation between the protective effects of the different treatment regimens (CsA, CsA plus IL-4, and CsA plus IL-10) and mRNA levels of type 1 cytokines (IFN-gamma, IL-2, and IL-12), type 2 cytokines (IL-4 and IL-10), or TGF-beta in the islet grafts.
Rabinovitch
A
A
Department of Medicine, University of Alberta, Edmonton, Canada.
Suarez-Pinzon
W L
WL
Sorensen
O
O
Rajotte
R V
RV
Power
R F
RF
D000818:Animals; D001786:Blood Glucose / Q000378:metabolism; D016572:Cyclosporine / Q000008:administration & dosage / Q000627:therapeutic use*; D016207:Cytokines / Q000096:biosynthesis / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D004359:Drug Therapy, Combination; D006085:Graft Survival / Q000187:drug effects*; D007166:Immunosuppressive Agents / Q000008:administration & dosage / Q000627:therapeutic use*; D016753:Interleukin-10 / Q000627:therapeutic use*; D015847:Interleukin-4 / Q000627:therapeutic use*; D016381:Islets of Langerhans Transplantation; D051379:Mice; D016688:Mice, Inbred NOD; D012333:RNA, Messenger / Q000378:metabolism
D001786:Blood Glucose; D016207:Cytokines; D007166:Immunosuppressive Agents; D012333:RNA, Messenger; D016753:Interleukin-10; D015847:Interleukin-4; D016572:Cyclosporine