牛白细胞介素-32β亚型的克隆与特性分析

Cloning and characterization of bovine interleukin-32 beta isoform.

作者信息

Jaekal Jun, Jhun Hyunjhung, Hong Jaewoo, Park Seungyoung, Lee Joongbok, Yoon Doyoung, Lee Siyoung, Her Erk, Yang Young, Rho Gyujin, Kim Soohyun

机构信息

Laboratory of Cytokine Immunology, Medical Immunology Center, Institute of Biomedical Science and Technology, Konkuk University, 1 Hwayang-dong, Gwangjin-gu, Seoul 143-701, Republic of Korea.

出版信息

Vet Immunol Immunopathol. 2010 Sep 15;137(1-2):166-71. doi: 10.1016/j.vetimm.2010.04.019. Epub 2010 May 7.

DOI:10.1016/j.vetimm.2010.04.019

PMID:20537728

Abstract

Interleukin-32 (IL-32) is a new cytokine produced mainly by T-cells, natural killer cells, and epithelial cells after stimulation with IL-2, IL-12 plus IL-18, and IFNgamma, respectively. IL-32 induces various proinflammatory cytokines such as TNFalpha, IL-1beta, IL-6, and IL-8 in monocytes and macrophages. In this study, we cloned bovine IL-32 cDNA from peripheral blood mononuclear cells (PBMC) of a Holstein (Bos Taurus). The bovine IL-32 cDNA has an entire open reading frame, encoding 171 amino acid residues and the deduced amino acid sequence has 27.5% identity with human IL-32 beta isoform. Recombinant bovine IL-32 protein was produced in E. coli and its molecular weight was approximately 26kDa. The biological activity of the bovine IL-32 was examined for cytokine induction using human monocytic THP-1 cells and bovine PBMC. The THP-1 cells responded to stimulation by recombinant bovine IL-32 and produced IL-8. Stimulation by recombinant bovine IL-32 induced mRNA for TNFalpha and IL-6 in bovine PBMC suggesting conservation of the biological activity and function in cattle.

摘要

白细胞介素-32（IL-32）是一种新的细胞因子，分别在受到IL-2、IL-12加IL-18以及IFNγ刺激后，主要由T细胞、自然杀伤细胞和上皮细胞产生。IL-32可诱导单核细胞和巨噬细胞产生多种促炎细胞因子，如TNFα、IL-1β、IL-6和IL-8。在本研究中，我们从荷斯坦牛（Bos Taurus）的外周血单个核细胞（PBMC）中克隆了牛IL-32 cDNA。牛IL-32 cDNA具有完整的开放阅读框，编码171个氨基酸残基，推导的氨基酸序列与人类IL-32β亚型具有27.5%的同一性。重组牛IL-32蛋白在大肠杆菌中产生，其分子量约为26kDa。使用人单核细胞THP-1细胞和牛PBMC检测了牛IL-32的细胞因子诱导生物活性。THP-1细胞对重组牛IL-32的刺激有反应并产生IL-8。重组牛IL-32的刺激在牛PBMC中诱导了TNFα和IL-6的mRNA，表明牛中生物活性和功能具有保守性。