犬腺病毒1型感染普通级比格犬中Ⅰ和Ⅲ型干扰素及其受体分子表达量的比较  

Comparison on the mRNA expression levels of typeⅠ and Ⅲ interferons and their receptors in CAV-1 infected dogs(Beagles)

作　　者：孟兴[1] 问婉欣 佟相慧 陈洪岩[1] 韩凌霞[1] MENG Xing;WEN Wan-xin;TONG Xiang-hui;CHEN Hong-yan;HAN Ling-xia(Division of Laboratory Animal and Comparative Medicine,State Key Laboratory of Veterinary Biotechnology,Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Heilongjiang Provincial Key Laboratory of Laboratory Animal and Comparative Medicine,Harbin 150069,China)

机构地区：[1]中国农业科学院哈尔滨兽医研究所实验动物与比较医学团队/兽医生物技术国家重点实验室黑龙江省实验动物与比较医学重点实验室,黑龙江哈尔滨150069

出　　处：《中国兽医科学》2021年第5期552-561,共10页Chinese Veterinary Science

基　　金：国家重点研发计划项目(2017YFD0501605)。

摘　　要：比较普通级比格犬感染犬腺病毒1型(CAV-1)后外周血和肝内Ⅰ型和Ⅲ型干扰素(IFN)及其受体分子的转录水平,同时评估CAV-2疫苗免疫对CAV-1感染试验的影响。对3只2月龄普通级比格犬后肢肌肉注射105.7TCID50CAV-1纯化病毒,在攻毒后12 d内记录体温变化,每日采集咽拭子和肛拭子,于攻毒后第3、6、9、12天采集前腔静脉血,利用常规PCR检测CAV-1和CAV-2核酸,利用间接免疫荧光试验(IFA)检测血清抗体效价。攻毒后第12天进行安乐死,采集心脏血和肝,利用定量PCR法检测血液和肝中IFN-α、IFN-β、IFNβR、IFN-λ1、IFNλ1R、IFNλ3和IFNλ3R等的转录水平。结果显示,31#犬的体温次日即升至39.7℃,出现便血和轻度腹泻,攻毒后第3天外周血中病毒核酸拷贝数达到峰值,随后在胸腺、腹股沟淋巴结、颌下淋巴结、肠系膜淋巴结和肝等组织中检测到CAV-1核酸,攻毒后第12天肝中IFN-λ1R的转录水平显著高于其他分子,IFA效价为1∶320。34#犬和35#犬的体温变化、临床症状、病毒血症、血清抗体效价和干扰素表达量均低于31#犬。实验早期犬的咽拭子和肛拭子中均检测到不同含量的CAV-2核酸。结果表明,CAV-1感染犬血液中的Ⅰ型和Ⅲ型干扰素表达规律不明显,肝是IFN-λ1主要的效应器官,同时CAV-2疫苗免疫对普通级实验犬的CAV-1感染研究具有一定的干扰。To compare the transcription levels of type Ⅰ and Ⅲ interferon(IFN) and their receptor molecules in peripheral blood and livers of conventional laboratory dogs(Beagles) infected with canine adenovirus type 1(CAV-1),and to access the effect of CAV-1 on the infection by CAV-2 vaccine.Three2-month-old Beagles were intramuscular injected with 105.7 TCID50 purified CAV-1.The body temperatures and clinical signs were recorded daily within 12 days(dpi).Pharyngeal swabs and anal swabs were collected daily,and venous blood in the anterior cavity was collected at 3,6,9 and 12 dpi.The nucleic acid of CAV-1 and CAV-2 genome was detected by PCR,and serum antibody titers were detected by indirect immunofluorescence assay(IFA).On 12 dpi, the dogs were euthanized and cardiac blood and livers were collected,and the transcription levels of IFN-α,IFN-β,IFNβR,IFN-λ1,IFN-λ1 R,IFN-λ3 and IFN-λ3 R in blood and liver were measured by quantitative PCR.The body temperature of Dog 31# increased to 39.7℃ on the next day,bloody stools and mild diarrhea.At 3 dpi,the virus nucleic acid copy number peaked in peripheral blood,and CAV-1 nucleic acid was detected in thymic,inguinal,submaxillary,mesenteric lymph nodes,liver and other tissues.On 12 dpi,the transcription level of IFN-λ1 R in the liver was significantly higher than that of other molecules,and the IFA titration was 1 ∶ 320.The changes of body temperature,clinical symptoms,viremia,serum antibody titer and interferon expression level were all lower in dog 34# and dog 35# than in dog 31#.Different amount of nucleic acid of CAV-2 virus were detected by PCR in pharyngeal swabs and anal swabs of all the 3 dogs at the early stage of CAV-1 inoculation.The expression of type Ⅰ and Ⅲ interferon in the blood of CAV-1 infected dogs was not obvious.Liver was the main effector organ of IFN-λ1,and CAV-2 immunization interferes the experimental study associated with canine infectious hepatitis in convention grade laboratory dogs.

关 键 词：比格犬 犬腺病毒1型 干扰素 干扰素受体 转录表达水平 

分 类 号：S852.659.1[农业科学—基础兽医学]