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猪传染性胃肠炎病毒TH-98株S基因
核酸疫苗的构建及其免疫效力
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任晓峰， 尹杰超， 司微， 李一经， 刘宝全
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(东北农业大学 动物医学院, 黑龙江 哈尔滨150030)
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摘要：
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利用DNA重组技术，将编码猪传染性胃肠炎病毒(TGEV) TH-98株的全长S基因和主要抗原位点的Sa片段分别插入真核表达载体pCI的多克隆位点中。应用酶切、PCR方法鉴定阳性重组子pCI-Sa和pCI-S。将昆明鼠随机分成A、B、C 3组（Ａ组为pCI-Sa免疫组、B组为pCI-S免疫组、C组为pCI载体对照组），每组35只，每只鼠肌肉注射100μg, 免疫3次，每次间隔２周。于初次免疫后第0、14、21、28、35、42d采血，用ELISA检测血清抗体动态变化，流式细胞仪检测外周血CD4+和CD8+T细胞数量变化。结果表明，重组真核表达载体均能诱导免疫鼠产生特异性的体液免疫及细胞免疫应答,而且pCI-Sa的免疫效果优于pCI-S。 |
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关键词：
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猪传染性胃肠炎病毒； S基因； 核酸疫苗； 构建； 免疫 |
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中图分类号：S 852．659.6 文献标识码：
A 文章编号：1673-4696(2006)03-0203-04 |
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Construction of nucleic acid vaccines containing S gene from
TGEV isolate TH-98 and their immune effect in mice |
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REN Xiao-feng, YIN Jie-chao, SI Wei, LI Yi-jing, LIU Bao-quan |
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(College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China) |
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Abstract: A full length spike(S) gene and a fragment Sa, encoding major antigenic sites, of transmissible gastroenteritis virus(TGEV) strain
TH-98 were cloned into the multiple cloning sites of the eukar-yotic expression vector pCI, respectively. The recombinant vaccines
pCI-Sa and pCI-S were verified with restriction digestion and PCR amplification. Kunming mice were divided randomly into Groups A, B and C of 35 mice each. Each mouse was injected intramuscularly with 100μg of
pCI-Sa, pCI-S and pCI, respectively. All groups were inoculated three times with the same volume at 2 week interval for improving immunization effect. On days 0, 14, 21, 28, 35 and 42
post-immunization, peripheral blood samples of the immunized mice were collected and subjected to antibody detection by ELISA, and quantitative analysis of CD4+ and CD8+ T cells by FACS. Results showed that both
pCI-Sa and pCI-S induced specific humoral and cellular immune responses, but
pCI-Sa was better than pCI-S. |
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Key words: transmissible gastroenteritis virus； S gene； nucleic acid vaccine； construction； vaccination |
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