马I型和II型白细胞介素-1受体cDNA的分子特征及功能表达

Molecular characterization and functional expression of equine interleukin-1 type I and type II receptor cDNAs.

作者信息

Kirisawa Rikio, Hashimoto Norikazu, Tazaki Mizuho, Yamanaka Hitoki, Ishii Risako, Hagiwara Katsuro, Iwai Hiroshi

机构信息

Department of Veterinary Microbiology, School of Veterinary Medicine, Rakuno Gakuen University, Bunkyoudai-Midorimachi 582, Ebetsu, Hokkaido 069-8501, Japan.

出版信息

Vet Immunol Immunopathol. 2006 Feb 15;109(3-4):219-31. doi: 10.1016/j.vetimm.2005.08.018. Epub 2005 Sep 19.

DOI:10.1016/j.vetimm.2005.08.018

PMID:16176839

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.

摘要

从脂多糖刺激的马外周血单核细胞生成的互补DNA（cDNA），分别使用源自人和小鼠白细胞介素-1受体I型（IL-1RI）和II型（IL-1RII）序列之间半保守区域的引物，来扩增和克隆马IL-1RI和IL-1RII。马IL-1RI和IL-1RII的5'和3'末端序列通过5'和3' cDNA末端快速扩增法进行扩增。推导的马IL-1RI氨基酸序列与人类、小鼠和大鼠序列的相似性分别为77%、64%和63%。预测的马IL-1RII氨基酸序列与人类、小鼠和大鼠序列的相似性分别为70%、60%和58%。在昆虫细胞中产生的重组马可溶性IL-1RI和IL-1RII与重组马IL-1α和IL-1β结合。此外，两种受体均以剂量依赖性方式抑制马IL-1α和IL-1β对A375细胞的生长抑制活性，表明目前的马IL-1RI和IL-1RII cDNA编码具有生物活性的蛋白质。