摘要
为了验证构建的重组腺病毒rAd-SRCR59-Fc能否在猪体内有效表达游离受体及其能否抑制PRRSV感染,将重组腺病毒注射断奶仔猪,用夹心ELISA检测SRCR59-Fc表达情况;选择PRRSV易感仔猪肌肉注射重组腺病毒rAd-SRCR59-Fc(4×109 TCID50/头),5 d后与JXA1株PRRSV人工感染猪同居饲养,定期测定体温、观察临床症状,采集血样和粪样进行PRRSV定量检测,对病死猪进行病理剖检和主要器官病毒载量测定。结果显示,重组腺病毒接种猪能表达SRCR59-Fc游离受体蛋白,并持续15 d左右;与感染对照组相比,rAd-SRCR59-Fc注射组首次高热时间和临床症状均推迟3 d,第5天~第13天临床症状评分显著低于感染组(P<0.01),至试验结束(25 d)仅有1头猪死亡;肺脏无明显肉眼病变,仅见少量坏死灶,病理切片显示肺泡壁轻微增厚;第3天~第13天病毒血症水平显著低于感染组(P<0.01),第5~第13天粪排毒量低于感染组(P<0.05),器官病毒载量均显著低于感染组(P<0.01)。研究表明CD163游离受体的重组腺病毒能有效阻断PRRSV毒株感染,可以作为PRRS预防或治疗性疫苗进一步研究。
To verify whether the constructed recombinant adenovirus rAd-SRCR59-Fc could express the soluble receptor and inhibit PRRSV infection invivo,piglets were inoculated with the rAd and SRCR59-Fc was detected by sandwich ELISA.PRRSV-na6 ve piglets were housed with one artificially infected pig of PRRSV JXA1 5 days later after inoculating muscularly with rAd-SRCR59-Fc(4×109 TCID50 each pig).Their temperatures and clinical symptoms were observed daily,and sera and fecal samples were collected regularly for PRRSV quantitative detection.The pigs died of PRRS were examined pathologically and the PRRSV loads in different tissues were detected by quantitative RT-PCR.Results showed that the rAd-inoculated pigs could express and secrete soluble receptor SRCR59-Fc for 15 days.The time for the first high fever and PRRS symptoms were delayed by 3 days in rAd-inoculated group compared to the challenge control group,and its symptom score during 5-13 days was significantly lower(P<0.01)than that in the challenge group with only one pig died at the end of the experiment(25 days later).Pigs in rAd-inoculated group did not show obvious gross or microscopic lesions in lung,and their viremia level was significantly lower(P<0.01)than that in the challenge group during 3-13 days,while fecal virus emission level was significantly lower(P<0.05)during 5-13 days.The viral loads in all tissues were obviously lower(P<0.01)than that in the challenge group.These data suggest that the rAd-delivered CD163 soluble receptor can block PRRSV infection obviously and considered for further research in PRRS prevention and control.
作者
吴植
夏文龙
郭长明
朱善元
孙怀昌
WU Zhi;XIA Wen-long;GUO Chang-ming;ZHU Shan-yuan;SUN Huai-chang(Jiangsu Key Laboratory for High-Tech Research and Development of Veterinary Biopharmaceuticals,Jiangsu Agri-Animal Husbandry Vocational College,Taizhou,Jiangsu,225300,China;College of Veterinary Medicine,Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses,Yangzhou University,Yangzhou,Jiangsu,225009,China)
出处
《动物医学进展》
北大核心
2020年第1期14-19,共6页
Progress In Veterinary Medicine
基金
江苏高校优势学科建设工程项目PAPD
江苏省农业科技自主创新资金项目[CX(14)2087]
江苏省自然科学基金项目(BK20151576)
关键词
猪繁殖与呼吸综合征病毒
重组腺病毒
可溶性病毒受体
抗病毒作用
Porcine reproductive and respiratory syndrome virus
recombinant adenovirus
soluble viral receptor
antiviral effect
