Cloning and characterization of recombinant rhesus macaque IL-10/Fcala-ala fusion protein: A potential adjunct for tolerance induction strategies

Academic Article

Abstract

  • The powerful anti-inflammatory and immunosuppressive activities of IL-10 make it attractive for supplemental therapy in translational tolerance induction protocols. This is bolstered by reports of IL-10-mediated inhibition of innate immunity, association of human stem cell and nonhuman primate (NHP) islet allograft tolerance with elevated serum IL-10, and evidence that systemic IL-10 therapy enhanced pig islets survival in mice. IL-10 has not been examined as adjunctive immunosuppression in NHP. To enable such studies, we cloned and expressed rhesus macaque (RM) IL-10 fused to a mutated hinge region of human IgG1 Fc to generate IL-10/Fcala-ala. RM IL-10/Fcala-ala was purified to ∼98% homogeneity by affinity chromatography and shown to be endotoxin-free (<0.008 EU/μg protein). The biological activity of IL-10/Fcala-ala was demonstrated by (1) costimulation of the mouse mast cell line, MC/9 proliferation in a dose-dependent fashion, (2) suppression of LPS-induced septic shock in mice and (3) abrogation of LPS-induced secretion of proinflammatory cytokines/chemokines in vitro and in vivo in NHP. Notably, RM IL-10/Fcala-ala had significantly greater potency than human IL-10/Fcala-ala and exhibited a circulating half-life of ∼14 days. The availability of this reagent will facilitate definitive studies to determine whether supplemental therapy with RM IL-10/Fcala-ala can influence tolerance outcomes in NHP. © 2007 Elsevier Ltd. All rights reserved.
  • Published In

  • Cytokine  Journal
  • Digital Object Identifier (doi)

    Author List

  • Asiedu C; Guarcello V; Deckard L; Jargal U; Gansuvd B; Acosta EP; Thomas JM
  • Start Page

  • 183
  • End Page

  • 192
  • Volume

  • 40
  • Issue

  • 3