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Receptor Engagement on Cells Expressing a Ligand for the Tolerance-Inducing Molecule OX2 Induces an Immunoregulatory Population That Inhibits Alloreactivity In Vitro and In Vivo¹

Reginald M. Gorczynski^2,*,, Kai Yu^* and David Clark^*,

^* University Health Network, Toronto, Canada; Departments of Surgery and Immunology, University of Toronto, Toronto, Canada; and Departments of Medicine, Pathology, and Molecular Medicine, Obstetrics and Gynecology, McMaster University, Hamilton, Ontario, Canada

Increased survival of C57BL/6 renal allografts following portal vein donor-specific pretransplant immunization of C3H mice is associated with increased expression of the molecule OX2 seen on host dendritic cells, along with a marked polarization in cytokine production from lymphocytes harvested from the transplanted animals, with preferential production of IL-4, IL-10, and TGF-ß on donor-specific restimulation in vitro, and decreased production of IL-2, IFN-, and TNF- compared with non-portal vein-immunized control transplanted mice. The increased renal allograft survival and the altered cytokine production are abolished by infusion of anti-mouse OX2 mAb (3B6). Infusion of a soluble OX2:Fc immunoadhesin can itself produce significant prolongation of xeno- and allografts in mice. We have used FITC-conjugated OX2:Fc to characterize cells expressing a ligand (OX2L) for OX2, and provide evidence that subpopulations of LPS-stimulated splenic macrophages, Con A-activated splenic T cells, and the majority (>80%) of TCR⁺ T cells express this ligand. We show below that F4/80⁺, OX2L⁺ splenic macrophages, admixed with OX2:Fc, represent a potent immunosuppressive population capable of causing more profound inhibition of alloreactivity in vitro or in vivo than that seen using either OX2:Fc or OX2⁺ (or OX2L⁺) cells alone. Immunoregulation by this OX2L⁺ population occurs in an MHC-restricted fashion.

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