Molecular characterization and functional expression of equine interleukin-1 type I and type II receptor cDNAs |
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Authors: | Kirisawa Rikio Hashimoto Norikazu Tazaki Mizuho Yamanaka Hitoki Ishii Risako Hagiwara Katsuro Iwai Hiroshi |
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Institution: | Department of Veterinary Microbiology, School of Veterinary Medicine, Rakuno Gakuen University, Bunkyoudai-Midorimachi 582, Ebetsu, Hokkaido 069-8501, Japan. r-kirisa@rakuno.ac.jp |
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Abstract: | cDNA generated from lipopolysaccharide-stimulated equine peripheral blood mononuclear cells was used to amplify and clone type I and type II equine interleukin-1 receptors (IL-1RI and IL-1RII) using primers derived from semi-conserved regions between human and mouse IL-1RI and IL-1RII sequences, respectively. 5' and 3' terminal sequences of equine IL-1RI and IL-1RII were amplified by 5' and 3' rapid amplification of cDNA ends. The deduced amino acid sequence of equine IL-1RI demonstrated 77, 64 and 63% similarity with human, mouse and rat sequences, respectively. The predicted amino acid sequence of equine IL-1RII demonstrated 70, 60 and 58% similarity with human, mouse and rat sequences, respectively. Recombinant equine soluble IL-1RI and IL-1RII produced in insect cells bound recombinant equine IL-1alpha and IL-1beta. Furthermore, both receptors suppressed the growth inhibitory activities of equine IL-1alpha and IL-1beta toward A375 cells in a dose-dependent manner, indicating that the present equine IL-1RI and IL-1RII cDNA encodes biologically active proteins. |
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