title: Cloning and characterization of recombinant rhesus macaque IL-10/Fc(ala-ala) fusion protein: a potential adjunct for tolerance induction strategies.
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/Fc(ala-ala). RM IL-10/Fc(ala-ala) was purified to approximately 98% homogeneity by affinity chromatography and shown to be endotoxin-free (<0.008 EU/microg protein). The biological activity of IL-10/Fc(ala-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/Fc(ala-ala) had significantly greater potency than human IL-10/Fc(ala-ala) and exhibited a circulating half-life of approximately 14 days. The availability of this reagent will facilitate definitive studies to determine whether supplemental therapy with RM IL-10/Fc(ala-ala) can influence tolerance outcomes in NHP.
authors.affiliation: Division of Transplant Immunology, Department of Surgery, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
authors.forename: C
authors.initials: C
authors.lastname: Asiedu
authors.forename: V
authors.initials: V
authors.lastname: Guarcello
authors.forename: L
authors.initials: L
authors.lastname: Deckard
authors.forename: U
authors.initials: U
authors.lastname: Jargal
authors.forename: B
authors.initials: B
authors.lastname: Gansuvd
authors.forename: E P
authors.initials: EP
authors.lastname: Acosta
authors.forename: J M
authors.initials: JM
authors.lastname: Thomas
mesh_terms: D000818:Animals; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects*; D003001:Cloning, Molecular; D004305:Dose-Response Relationship, Drug; D006085:Graft Survival / Q000187:drug effects / Q000276:immunology; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000235:genetics / Q000276:immunology / Q000494:pharmacology*; D007074:Immunoglobulin G / Q000235:genetics / Q000276:immunology / Q000494:pharmacology; D050417:Insulin-Secreting Cells / Q000276:immunology / Q000637:transplantation; D016753:Interleukin-10 / Q000235:genetics / Q000276:immunology / Q000493:pharmacokinetics / Q000494:pharmacology*; D008070:Lipopolysaccharides / Q000633:toxicity; D008253:Macaca mulatta; D008407:Mast Cells / Q000276:immunology*; D051379:Mice; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000493:pharmacokinetics / Q000494:pharmacology*; D012772:Shock, Septic / Q000139:chemically induced / Q000188:drug therapy* / Q000276:immunology; D013552:Swine; D023001:Transplantation Tolerance / Q000187:drug effects*; D014183:Transplantation, Heterologous
chemical_list: D007141:Immunoglobulin Fc Fragments; D007074:Immunoglobulin G; D008070:Lipopolysaccharides; D011993:Recombinant Fusion Proteins; D016753:Interleukin-10