Cloning and characterization of recombinant rhesus macaque IL-10/Fcala-ala fusion protein: A potential adjunct for tolerance induction strategies
| Autor: | Judith M. Thomas, Lindsey A. Deckard, Uuganbayar Jargal, Balgansuren Gansuvd, E.P. Acosta, Vincenzo Guarcello, Clement Asiedu |
|---|---|
| Rok vydání: | 2007 |
| Předmět: |
Lipopolysaccharides
Swine Recombinant Fusion Proteins Transplantation Heterologous Immunology Pharmacology Biochemistry Cell Line Proinflammatory cytokine Mice In vivo Insulin-Secreting Cells Animals Humans Immunology and Allergy Mast Cells Cloning Molecular Molecular Biology Cell Proliferation Dose-Response Relationship Drug biology Graft Survival Hematology biology.organism_classification Macaca mulatta Shock Septic Fusion protein In vitro Immunoglobulin Fc Fragments Interleukin-10 Rhesus macaque Tolerance induction Interleukin 10 Immunoglobulin G Transplantation Tolerance Stem cell |
| Zdroj: | Cytokine. 40:183-192 |
| ISSN: | 1043-4666 |
| DOI: | 10.1016/j.cyto.2007.09.008 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect