鸡β-防御素-1对鸡传染性法氏囊病病毒VP2基因DNA疫苗的免疫佐剂作用

畜牧兽医学报 ›› 2011, Vol. 42 ›› Issue (3): 402-408.doi:

鸡β-防御素-1对鸡传染性法氏囊病病毒VP2基因DNA疫苗的免疫佐剂作用

张辉华¹*，杨小梅²，谢青梅²，马静云²，罗艳娜^1,2，曹永长³，毕英佐^2,3

1.佛山科技学院生命科学院,佛山 528231；2.华南农业大学动物科学学院,广州 510642； 3.中山大学生命科学学院,广州 510275

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2011-03-25 发布日期:2011-03-25
通讯作者: 张辉华

The Adjuvant Effects of Chicken β-defensin-1 on Infectious Bursal Disease Virus VP2 Gene DNA Vaccine

ZHANG Hui-hua¹*, YANG Xiao-mei², XIE Qing-mei², MA Jing-yun², LUO Yan-na^1,2, CAO Yong-chang³, BI Ying-zuo^2,3

1. College of Life Sciences, Foshan University, Foshan 528231, Guangdong, China;2. College of Animal Science, South China Agricultural University, Guangzhou 510642, China;3. College of Life Sciences, Sun Yatsen University, Guangzhou 510275, China

Received:1900-01-01 Revised:1900-01-01 Online:2011-03-25 Published:2011-03-25

摘要/Abstract

摘要： 本文旨在研究鸡β-防御素-1（AvBD1）对鸡IBDV VP2基因DNA疫苗的免疫佐剂作用。通过基因工程技术，构建真核表达质粒pcDNA3.1(+)-AvBD1与pcDNA3.1(+)-VP2。14日龄岭南黄肉鸡随机分成5组，分别免疫PBS、空质粒载体pcDNA3.1(+)、重组质粒pcDNA3.1(+)-VP2、重组质粒pcDNA3.1(+)-AvBD1与pcDNA3.1(+)-VP2联合免疫、IBD弱毒苗。收集免疫后不同时期的外周血血清样本，用ELISA方法检测血清中IBDV VP2特异性抗体水平，通过流式细胞仪检测CD3⁺、CD4⁺与CD8⁺T淋巴细胞的含量。结果表明，联合免疫组（即pcDNA3.1(+)AvBD1与pcDNA3.1(+)-VP2共同免疫）VP2特异性抗体水平显著高于其他组；单独免疫质粒pcDNA3.1(+)-VP2组VP2特异性抗体水平与免疫IBD弱毒苗组相当，显著高于PBS组与空质粒载体pcDNA3.1(+)组。在二免后第7天，联合免疫组外周血中CD3⁺、CD4⁺与CD8⁺T淋巴细胞的含量也显著高于单独免疫质粒pcDNA3.1(+)-VP2组，其它时间无显著性差异。IBDV攻毒后的保护率也是联合免疫组高于单独免疫pcDNA3.1(+)-VP2组。这些结果表明，鸡AvBD1可以作为一种免疫佐剂用于增强IBDV VP2基因DNA疫苗的免疫力。

Abstract: The aim of present study was to investigate the adjuvant effects of chicken β-defensin-1 (also named avian β-defensin-1, AvBD1) on DNA vaccine encoding chicken infectious bursal disease virus (IBDV) VP2 gene. The eukaryotic expression vectors pcDNA3.1(+)-AvBD1 and pcDNA3.1(+)-VP2 were constructed. Five groups of 14-day-old chickens were intramuscularly injected with PBS, empty vector pcDNA3.1(+), pcDNA3.1(+)-VP2, a mixtures of pcDNA3.1(+)-AvBD1 and pcDNA3.1(+)-VP2, and IBD attenuated vaccine, respectively. Serum of chicken in each group was collected from the peripheral blood at different time post inoculation, and VP2specific antibody was measured by ELISA. Content of CD3⁺, CD4⁺ and CD8⁺ T-lymphocytes in each group of chickens were tested by cytospectrometry. Results showed that VP2-specific antibody level of group immunized with a mixtures of pcDNA3.1(+)-AvBD1 and pcDNA3.1(+)-VP2 was the highest among all the groups. The VP2-specific antibody level of group immunized with pcDNA3.1(+)-VP2, as well as group immunized with IBD attenuated vaccine, was significantly higher than that of the group immunized with PBS and empty vector pcDNA3.1(+). The percentages of CD3⁺, CD4⁺ and CD8⁺ T-cell subtypes between groups of pcDNA3.1(+)-VP2 and pcDNA3.1(+)-VP2 combination with pcDNA3.1(+)-AvBD1 was significantly different (P<0.05), the latter was higher, at 7 days post boost. The protection from IBD challenged by immunized chickens with DNA vaccines encoding IBDV VP2 gene alone was lower than that by immunized with a mixtures of pcDNA3.1(+)-AvBD1 and pcDNA3.1(+)-VP2. The results indicated that AvBD1 can be used as an adjuvant to enhance the IBDV VP2 DNA vaccine immunity.

张辉华;杨小梅;谢青梅;马静云;罗艳娜;曹永长;毕英佐;. 鸡β-防御素-1对鸡传染性法氏囊病病毒VP2基因DNA疫苗的免疫佐剂作用[J]. 畜牧兽医学报, 2011, 42(3): 402-408.

ZHANG Hui-hua;YANG Xiao-mei;XIE Qing-mei;MA Jing-yun;LUO Yan-na;CAO Yong-chang;BI Ying-zuo;. The Adjuvant Effects of Chicken β-defensin-1 on Infectious Bursal Disease Virus VP2 Gene DNA Vaccine[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2011, 42(3): 402-408.