We present here the first report of a metalloporphyrin-based antioxidant that can prevent or delay the onset of autoimmune diabetes. Type 1 diabetes is an autoimmune process whereby T-cells recognize pancreatic β-cell antigens and initiate a leukocyte infiltrate that produces proinflammatory cytokines and reactive oxygen species (ROS), ultimately leading to β-cell destruction. Because islet β-cells have a reduced capacity to scavenge free radicals, they are very sensitive to ROS action. Metalloporphyrin-based superoxide dismutase (SOD) mimics scavenge ROS and protect cells from oxidative stress and apoptosis. To investigate the effect of SOD mimics and the role of oxidative stress in the development of autoimmune diabetes in vivo, we used a diabetogenic T-cell clone, BDC-2.5, to induce rapid onset of diabetes in young nonobese diabetic-severe combined immunodeficient mice (NOD.scid). Disease was significantly delayed or prevented altogether by treatment of recipient mice with an SOD mimic, AEOL-10113, before transfer of the BDC-2.5 clone. To investigate the mechanisms of protection, in vitro assays for T-cell proliferation and γ-interferon (IFN-γ) production were carried out using the T-cell clone BDC-2.5. We found that the SOD mimic significantly inhibited antigen-presenting cell-dependent T-cell proliferation and IFN-γ production in vitro. In addition, pretreatment of lipopolysaccharide (LPS)-stimulated peritoneal macrophages with SOD mimic inhibited the LPS-dependent increase in TNF-α as well as the NADPH oxidase-dependent release of superoxide. Finally, this compound protected NIT-1 insulinoma cells from interleukin-1β and alloxan cytotoxicity in vitro.
Address correspondence and reprint requests to Jon D. Piganelli, Children’s Hospital of Pittsburgh, 3460 5th Ave., Rangos Research Building, Pittsburgh, PA 15213. E-mail:.
Received for publication 2 May 2001 and accepted in revised form 25 October 2001.
J.D.P. and S.C.F. contributed equally to this work.
J.C. and B.D. hold stock in and have received consulting fees and laboratory funds from Incara Pharmaceuticals.
CM, complete medium; ConA, concanavalin A; ELISA, enzyme-linked immunosorbent assay; HBSS, Hanks’ balanced salt solution; IFN-γ, γ-interferon; IL, interleukin; LPS, lipopolysaccharide; MAPK, mitogen-activated protein kinase; NF-κB, nuclear factor-κB NO·, nitric oxide; O2·−, superoxide; ·OH, hydroxyl radical; ONOO−, peroxynitrite; PC, peritoneal macrophage; PMA, phorbol myristate acetate; PMN, polymorphonuclear leukocyte; RNI, reactive nitrogen intermediate; ROS, reactive oxygen species; SOD, superoxide dismutase; TNF-α, tumor necrosis factor-α.