1. Cloning and characterization of rhesus IL-18 binding protein, a natural antagonist to IL-18.
- Author
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Yellayi S, Braun S, Domingues HG, Kityatana N, Murali R, Johnson RP, Mansfield K, Westmoreland SV, and O'Neil SP
- Subjects
- Amino Acid Sequence, Animals, Atherosclerosis metabolism, Base Sequence, Cloning, Molecular, Humans, Immunoglobulin G metabolism, Immunohistochemistry, Intercellular Signaling Peptides and Proteins chemistry, Interferon-gamma biosynthesis, Interleukin-18 pharmacology, Lipopolysaccharides pharmacology, Mice, Molecular Sequence Data, Molecular Weight, Protein Transport drug effects, Recombinant Fusion Proteins metabolism, Sequence Analysis, Protein, Spleen cytology, Spleen drug effects, Spleen metabolism, Intercellular Signaling Peptides and Proteins genetics, Intercellular Signaling Peptides and Proteins metabolism, Interleukin-18 antagonists & inhibitors, Macaca mulatta genetics
- Abstract
IL-18 is a proinflammatory cytokine that is important for host defense, but is also involved in the pathogenesis of a number of disease processes, ranging from autoimmune disorders to atherosclerosis. IL-18 binding protein (IL-18BP) is a constitutively expressed glycoprotein that specifically neutralizes the effects of IL-18, resulting in decreased production of IFN-gamma and reduction in Th1 immune responses. In this study we cloned and sequenced a full-length cDNA of the rhesus IL-18BP (RhIL-18BP) from the spleen of rhesus macaques (Macaca mulatta) and compared its nucleotide and amino acid sequences to the functional murine and human IL-18BP orthologues. In addition, we fused RhIL-18BP to the Fc portion of human IgG1 to make recombinant RhIL-18BP x Fcgamma1 in order to facilitate its detection by Western blot analysis and determined the approximate molecular weight of RhIL-18BP x Fcgamma1 to be 66 kD. With this fusion protein, we showed that RhIL-18BP was functional and could significantly reduce murine IL-18 and LPS-induced IFN-gamma production by murine splenocytes. Furthermore, we demonstrated the expression of IL-18BP in atherosclerotic lesions in a rhesus model of atherosclerosis, underscoring the need to fully understand the role of this protein as a primary negative regulator of IL-18 in multiple disease processes., (Copyright 2010 Elsevier Ltd. All rights reserved.)
- Published
- 2010
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