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表达猪瘟病毒石门株E0和/或E2基因重组非复制型腺病毒疫苗的免疫保护试验
张永科1, 杨玉艾1, 王养会1
西北农林科技大学 动物科技学院
摘要:
用已构建好的表达猪瘟病毒石门株E0基因的重组腺病毒pAd-E0、表达猪瘟病毒石门株E2基因的重组腺病毒pAd-E2以及联合表达猪瘟病毒石门株E0-E2基因的重组腺病毒pAd-E0-E2,肌肉和皮下接种免疫商品猪2次(间隔20 d),同时设非重组腺病毒PAD-CMV、常规猪瘟疫苗和空白对照组。二免21 d后进行CSFV石门株强毒超致死量(1 000倍TCID50)攻击,研究表达猪瘟保护性抗原基因重组腺病毒疫苗的免疫效果。结果表明,pAd-E0、pAd-E2和pAd-E0-E2免疫组猪的死亡率分别为50%,25%和25%,而常规猪瘟兔化弱毒疫苗免疫组的死亡率为40%,pAd-CMV免疫组和空白对照组猪全部死亡;在攻毒前,重组腺病毒免疫组抗体水平普遍较低,商品化疫苗免疫组能100%检测到抗体,而非重组腺病毒和空白对照没有检测到抗体;攻毒后pAd-E0、pAd-E2和pAd-E0-E2试验组中分别有75%,50%和50%猪体温超过40 ℃,常规苗试验组80%体温超过40℃,而PAD-CMV组和空白对照组所有试验猪体温均超过40 ℃,表明重组腺病毒疫苗具有一定的保护效果。表明,仅含有E0基因的重组腺病毒pAd-E0的免疫保护效果和常规疫苗还有距离;含有E2基因和E0-E2基因的重组腺病毒的免疫保护效果不亚于常规疫苗。
关键词:  猪瘟病毒  重组腺病毒  E0基因  E2基因  保护效果
DOI:
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基金项目:陕西省科技攻关项目(2003K02-G11-01);国家自然科学基金项目(30471290)
Immunological characterization of the replication-defective recombinant adenovirus expressing E0 and/or E2 antigen of classical swine fever virus Shimen strain
Abstract:
The commercially available pig’s susceptibility to CSFV was determined 21 days after twice immunizations through subcutaneous and intramuscular routes by three recombinant adenoviruses which contained the Classical Swine Fever Virus (CSFV) E0 and/orE2 gene,and set pigs inoculated non-recombinant adenovirus and DMEM as negative control.The mortality rate of pigs vaccinated with pAd-E0,pAd-E2 and pAd-E0-E2 were 50 percent,25 percent and 25 percent,respectively,while 40 percent of animals vaccinated with commercial vaccine were dead,and all animals vaccinated with the non-recombinant human adenovirus and blanks were dead.Before challenge,low antibodies to CSFV were detected in all pigs vaccinated with recombinant adenoviruses.High antibodies were detected in all animals vaccinated with commercial vaccine,but none in blanks.The number of pigs which had a temperature above 40 ℃ (indicative of fever) were tested every day after challenge.Results showed 75 percent of animals (3/4) vaccinated with pAd-E0,50 ercent of animals (2/4) vaccinated with pAd-E2,and 50 percent of animals (4/8) vaccinated with pAdE0-E2 developed temperatures above 40 ℃;80 percent of pigs (4/5) vaccinated with commercial vaccine developed temperatures above 40 ℃,while all blanks developed temperatures above 40 ℃.The analysis of clinical symptom showed that the recombinant adenoviruses could offer protection to CSFV.The above results showed that the recombinant adenovirus contained CSFV E0 gene could offer less protection for pigs than commercial vaccine,but recombinant adenoviruses contained CSFV E2 could offer more protection than commercial vaccine.
Key words:  classical swine fever virus,recombinant-defective adenovirus,E0 gene,E2 gene,protection effect