遗传 ›› 2015, Vol. 37 ›› Issue (1): 63-69.doi: 10.16288/j.yczz.2015.01.009

• 研究报告 • 上一篇    下一篇

聚肌胞免疫刺激下仔猪外周血单核细胞的基因表达分析

王彦平1, 王怀中1, 郭建凤1, 王海飞2, 刘剑锋2, 王继英1   

  1. 1. 山东省农业科学院畜牧兽医研究所,山东省畜禽疫病防治与繁育重点实验室,济南 250100;
    2. 中国农业大学动物科技学院,农业部畜禽遗传育种与繁殖重点实验室,北京 100193
  • 收稿日期:2014-03-10 修回日期:2014-05-05 出版日期:2015-01-20 发布日期:2015-01-20
  • 通讯作者: 王继英,博士,副研究员,研究方向:猪的遗传育种。E-mail: jnwangjiying@163.com E-mail:jnwangjiying@163.com
  • 作者简介:王彦平,硕士,助理研究员,研究方向:猪的遗传育种。E-mail: wangyanping03@163.com
  • 基金资助:
    国家自然科学基金项目(编号:31201779),山东省农业良种工程项目(编号:2011LZ13-01, 02),山东省优秀中青年科学家科研奖励基金项目(编号:BS2013SW026)和山东省农业产业技术体系生猪创新团队建设项目(编号:SDAIT-06-011-03)资助

Gene expression analysis of porcine peripheral blood mononuclear cell in response to immune stimulation of Poly I:C

Yanping Wang1, Huaizhong Wang1, Jianfeng Guo1, Haifei Wang2, Jianfeng Liu2, Jiying Wang1   

  1. 1. Shandong Key Laboratory of Animal Disease Control and Breeding, Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences, Jinan 250100, China; 2. Key Laboratory of Animal Genetics, Breeding and Reproduction, Ministry of Agriculture, College of Animal Science and Technology,China Agricultural University, Beijing 100193, China
  • Received:2014-03-10 Revised:2014-05-05 Online:2015-01-20 Published:2015-01-20

摘要: 聚肌胞(Poly I:C)是一种天然双链RNA(Double strand RNA, dsRNA)的拟似物,能够模拟病毒感染后所形成的dsRNA及刺激机体产生抗病毒免疫反应。文章以抗病力存在差异的大蒲莲和长白仔猪为研究对象,分离外周血单核细胞(Peripheral blood mononuclear cell, PBMC),在20 μg/mL的Poly I:C的免疫刺激下体外培养24 h,对影响免疫应答过程中的7个细胞因子(IRF3IL6IL8IL10TNFαIFNγIFNα)和3个模式识别受体(TLR3TLR4RIG1)利用实时荧光定量PCR检测Poly I:C免疫刺激组相对于对照组的基因表达变化倍数。结果表明:检测的大部分细胞因子和受体(6个)表达量变化倍数很大,其中3种白细胞介素IL6IL8IL10免疫刺激变化倍数最大,平均变化倍数分别为20.71、10.87和5.18倍。对不同个体和品种间的比较发现,不仅大蒲莲和长白两品种间(大蒲莲猪的变化倍数平均高于长白猪)而且同品种的3头全同胞仔猪间对Poly I:C免疫刺激的应答也存在较大的变化。文章利用Poly I:C体外模拟dsRNA对PBMC的感染,为下一步筛选仔猪对Poly I:C刺激的免疫应答基因及鉴定大蒲莲猪特殊的抗性基因奠定了基础。

关键词: 聚肌胞, 外周血单核细胞, 细胞因子, 模式识别受体

Abstract: Polyinosinic-polycytidylic acid (Poly I:C) is an analogue of natural double strand RNA (dsRNA), which can simulate the viral dsRNA and stimulate the immune response. In the present study, peripheral blood mononuclear cells (PBMC) were isolated from piglets of Dapulian and Landrace with different disease resistance, and stimulated with 20 μg/mL Poly I:C for 24 hours in vitro culture. The expression of several cytokines (IL6, IL8, TNFα, IL10, IRF3, IFNα and IFNγ) and three pattern recognition receptors (TLR3, TLR4 and RIG1) was determined by qRT-PCR. The results showed that, most of the cytokines or receptors had obvious expression change compared with the control (without Poly I:C stimulation), especially the three cytokine genes IL6, IL8 and IL10, whose average expression change times were 20.71, 10.87 and 5.18, respectively. Expression comparison between breeds and among individuals of the same breed indicated that there was obvious difference not only between Dapulain and Landrace (Dapulain higher than Landrace) but also among the three individuals of the same breed. Our study simulated the infection of dsRNA to host cells using Poly I:C, and provided experimental foundation for further study on selecting the immune genes in response to Poly I:C stimulation and identifying the unique disease-resistance genes of Dapulian.

Key words: Polyinosinic-polycytidylic acid (Poly I:C), peripheral blood mononuclear cell (PBMC), cytokine, pattern recognition receptors