摘要
将构建的鹅细小病毒（GPV）VP3基因疫苗（pcDNA-GPV-VP3）转染鹅胚成纤维细胞（GEF），每隔12h检测VP3蛋白的表达情况，同时以不同剂量分别通过基因枪轰击和肌肉注射免疫28日龄健康雏鹅，于免疫后第3、7、14、21、28、35、49、63、77和105d采血，进行淋巴细胞增殖试验（MTT法）。结果，转染后第24h即可在GEF中检测到GPVVP3蛋白，第60h表达量达到高峰。试验组鹅外周血T淋巴细胞D490nm值在免疫后第35d达到最大。肌肉注射组和基因枪组免疫后第14～63d、第7～63d的D490nm值分别与PBS对照组差异板显著；肌肉注射组及基因枪组免疫后第21～49d的D490nm值显著高于弱毒疫苗对照组；肌肉注射组及基因枪组免疫后第14～63d的D490nm值板显著高于空质粒对照组。表明，基因枪轰击和肌肉注射pcDNA—GPV—VP3均能诱导雏鹅产生良好的细胞免疫应答。
A plasmid pcDNA-GPV-VP3 was transfected into goose embryo fibroblast cells（GEFs）,and the expression of VP3 protein was detected every 12 hours. 28-day-old healthy goslings were immunized with different dosages of pcDNA-GPV-VP3 by gene gun bombardment and intramuscular injection（i. m. ）, respectively. Blood samples collected on day 3,7,14,21,28,35,49,63,77 and 105 post-immunization （PI） were measured by T-lymphocyte proliferation test. In results,the VP3 gene of goose parvovirus（GPV） was successfully expressed in GEFs at hour 24 PI and the expression reached peek at hour 60 PI. D490nm of T-lymphocyte cells from the immunized goslings reached peak on day 35 PI. D490nm in the i. m. and gene gun bombardment groups were very significantly higher than that in the PBS control group on day 14-63 and 7-63 PI respectively,and significantly higher than that in the GPV attenuated vaccine group on day 14 -63 PI and that in the empty plasmid group on day 21-49 PI. The results showed that the good cellular immunity effects were induced with pcDNA-GPV-VP3 by i. m. and gene gun bombardment.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2007年第5期401-405,共5页
Chinese Veterinary Science
基金
国家科技攻关重大项目(2004BA901A03)
教育部"新世纪优秀人才支持计划"项目(NCET-04-0906)
四川省基础研究重大项目(04JY029-006-1)
四川省重点建设学科项目(SZD0418)
