传染性法氏囊病病毒对鸡法氏囊组织中干扰素和p53转录的影响  

Differential modulation of interferon and p53 mRNA expression in chickens infected with infectious bursal disease virus

作　　者：欧阳伟[1,2,3] 王永山[1] 刘华洁[1,2] 杜希宁[1,4] 张海彬[2] 

机构地区：[1]江苏省农业科学院兽医研究所/农业部兽用生物制品工程技术重点实验室/国家兽用生物制品工程技术研究中心,南京210014 [2]南京农业大学动物医学院,南京210095 [3]江苏省动物重要疫病与人兽共患病防控协同创新中心,江苏扬州225009 [4]南京师范大学生命科学学院,南京210046

出　　处：《中国兽医学报》2015年第6期862-867,共6页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31272537;31302070);江苏省自然科学基金资助项目(BK2012787);江苏省农业科技自主创新基金资助项目(CX(13)5032)

摘　　要：为了分析传染性法氏囊病病毒(IBDV)感染对鸡体抗病毒天然免疫能力的影响,本研究应用IBDV弱毒(B87)和强毒(QL/12)分别感染4周龄SPF鸡,每隔12h(12~84h)取3只鸡的法氏囊组织作为1个pool,用荧光定量PCR(qRT-PCR)检测其感染前后干扰素(IFN)和p53的mRNA水平,同时分析法氏囊组织的病理变化和病毒载量。结果显示:在QL/12组,IFN-α和IFN-βmRNA水平随着时间的增加逐渐升高,48h达到峰值,随后降低;IFN-γmRNA水平随着时间的增加逐渐升高,60h达到峰值,随后逐渐降低;p53mRNA含量迅速升高,60h达到峰值,随后降低。在B87组,IFN-αmRNA水平先降低后升高(48h达到最低值),而IFN-βmRNA水平变化不规则,72h含量最高;IFN-γmRNA水平先逐渐升高后降低(72h达到峰值);p53mRNA含量变化较小,36h含量最高。QL/12组诱导法氏囊组织产生的IFN mRNA(IFN-α、IFN-β和IFN-γmRNA)以及p53mRNA水平均显著高于B87组。病理组织学检查,QL/12组法氏囊淋巴滤泡呈不同程度的损伤,而B87组法氏囊组织未见明显的组织病变。qRT-PCR检测,QL/12组法氏囊组织中病毒含量48h达到峰值(8.4×106拷贝/mg),而B87组法氏囊组织中病毒含量72h达到峰值(6.6×104拷贝/mg)。本研究结果表明IBDV感染严重干扰了鸡体IFN和p53介导的抗病毒天然免疫反应。To analyze the antiviral innate immunity of chickens infected with IBDV,four-week-old specific pathogen free chickens were inoculated intraocularly with intermediate virulent strain(B87)and a very virulent variant strain(QL/12).The mRNA transcription of type I interferon(IFN-αand IFN-β),Th1cytokines(IFN- γ),p53 mRNA and virus load in bursal tissues for the first 84 hours after infection were quantified by real-time quantitative reverse transcription polymerase chain reaction(RT-PCR),respectively.The results showed that,in group QL/12,IFN-αand IFN-β mRNA level increased gradually with the time,reached the peak at 48 h,then decreased;IFN-α mRNA level increased gradually with the time,reached the peak at 60 h,then decreased gradually;P53mRNA content increased rapidly,reached the peak at 60 h,then decreased.In the B87 group,the level of IFN-αmRNA increased firstly and then decreased(the lowest value at 48h),and changes of IFN-βmRNA level was irregular,highest content at 72h;IFN-αmRNA levels increased gradually firstly then decreased(peak at 72h);change of P53 mRNA was not obvious,with the highest at 36 h.In QL/12 group,bursal tissue IFN mRNA(IFN-α,IFN-β0and IFN-γmRNA)and P53 mRNA levels were significantly higher than those in group B87.bursal lymphoid follicles of QL/12 group showed different degrees of injury,while group B87 bursal tissue was no obvious lesions.in QL/12 group,the virus content of bursa of Fabricius reached peak value(8.4×106 copies/mg)at 48 h,and the B87 group at 72h(6.6×104 copies/mg).QL/12 induced more notable damage of bursal follicular compared to B87.Virus load was significantly higher(P<0.05)in bursas obtained from QL/12-inoculated chickens than that from B87-inoculated chickens.The results provided direct experimental eveidences that IBDV infection results in the abnormal expression of IFN-α,IFN-β,IFN-γand P53,which is very important for the understanding IBDV interfering with antiviral innate immunity in chickens.

关 键 词：传染性法氏囊病病毒(IBDV) 干扰素 P53 天然免疫 

分 类 号：S852.65[农业科学—基础兽医学]