猪瘟病毒感染猪细胞因子和趋化因子的动态变化  

Kinetics of Cytokines and Chemokines of Pigs Infected with Classical Swine Fever Virus

作　　者：李素[1,2] 王惟[1,2] 张淑琴[3] 董鹏[3] 朱妍[2] 郭焕成[2] 涂长春[2] 

机构地区：[1]吉林大学畜牧兽医学院,长春130062 [2]军事医学科学院军事兽医研究所,长春130062 [3]中国农业科学院左家特产研究所,吉林吉林132109

出　　处：《中国生物制品学杂志》2010年第10期1038-1042,1049,共6页Chinese Journal of Biologicals

基　　金：国家重点基础研究发展计划(973)项目(2005CB523200);国家"十一五"科技支撑计划项目(2006BAD06A03)

摘　　要：目的分析猪瘟病毒(Classical swine fever virus,CSFV)感染猪细胞因子和趋化因子的动态变化。方法将健康猪随机分成2组:感染组和对照组,感染组经颈部肌肉注射CSFV石门株1×103TCID50/头,对照组注射等体积的生理盐水。感染后每日观察实验猪的临床症状。分别于感染前和感染后4、6、8d采血,分离外周血白细胞和血清,应用实时荧光定量RT-PCR检测外周血白细胞IL-1、IL-4、IL-6、IL-8、IL-10、IFNγ、TNF-α和干扰素诱导蛋白10(Interferon inducible protein10,IP-10)mRNA的转录水平及全血中的病毒载量;应用ELISA法检测血清中各细胞因子的含量。结果 CSFV感染后,猪的临床症状分值逐渐升高,体温在第3天达最高,然后下降;白细胞总体呈明显下降趋势;细胞因子IL-1、IL-4、IL-6、TNF-α和IP-10的mRNA转录水平上调,IL-10的mRNA转录水平下调,IFNγ的mRNA转录水平基本未发生变化;CSFV感染猪在感染初期和症状明显期(4d和6d)的外周血的病毒载量呈明显上升趋势,而在濒死期(8d)病毒载量有所下降;CSFV感染后猪血清中的TNF-α含量上升,IL-4、IL-6、IL-8、IL-10和IFNγ的含量下降。结论猪瘟病毒感染猪TNF-α和IL-10 mRNA的转录水平与血清中的细胞因子含量变化趋势一致,而其他细胞因子mRNA的转录水平与血清中含量变化并不一致。Objective To analyze the kinetics of cytokines and chemokines of pigs infected with classical swine fever virus （CSFV）. Methods Healthy pigs were randomly divided into test and control groups. Each pig in test groups was infected i.m. with CSFV Shimen strain at a dosage of 1 × 103 TCID50, while that in control group with an equal volume of physiological saline. Blood samples were collected before and 4, 6 and 8 d after challenge with CSFV respectively for separation of peripheral blood leukocytes and sera. The transcription levels of IL-1, IL-4, IL-6, IL-8, IL-10, IFNγ, TNF-α and IP-10 mRNAs in peripheral blood leukocytes as well as virus loading in blood were determined by real-time fluorescent quantitative RT-PCR, while the cytokine content in sera by ELISA. Results After infection with CSFV, the score for clinical signs of pigs increased gradually, while the body temperature reached a peak value on day 3 then decreased. The leukocyte level showed a decreasing tendency in general. The transcription levels of IL-1, IL-6 and IP-10 mRNAs were up-regulated, while that of IL-10 mRNA was down-regulated. However, the transcription level of IFNγ showed no significant change. The virus loading in peripheral blood of pigs increased significantly at early stage of infection and the stage in which obvious symptoms appeared （4 d and 6 d after infection）, while decreased at dying stage （8 d after infection）. The TNF-α content in sera of pigs after CSFV infection increased, while IL-4, IL-6, IL-8, IL-10, and IFNγ contents decreased. Conclusion The change tendencies of transcription levels of TNF-α and IL-10 mRNAs in pigs after infection with CSFV were consistent, while those of other cytokines were inconsistent with those of cytokine contents in sera.

关 键 词：猪瘟病毒 细胞因子 趋化因子 荧光定量RT-PCR 酶联免疫吸附测定 

分 类 号：S852.65[农业科学—基础兽医学]