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阿留申水貂病毒强毒株全基因变异DNA疫苗的构建及免疫原性分析

Construction and Immunogenicity Analysis of Whole-Gene Mutation DNA Vaccine of Aleutian Mink Virus Isolated Virulent Strain.

作者信息

Liu Dongxu, Li Jianming, Shi Kun, Zeng Fanli, Zong Ying, Leng Xue, Lu Huijun, Du Rui

机构信息

1 College of Chinese Medicine Material, Jilin Agricultural University , Changchun, People's Republic of China .

2 College of Animal Science and Technology, Jilin Agricultural Science and Technology University , Jilin, People's Republic of China .

出版信息

Viral Immunol. 2018 Jan/Feb;31(1):69-77. doi: 10.1089/vim.2017.0044. Epub 2017 Aug 22.

Abstract

Aleutian mink disease (AD) is a chronic viral infection that causes autoimmune disorders in minks and presents a significant economic burden on mink farming. Despite the substantial challenges presented by AD, no effective vaccine is available and only partial protection has been achieved. We constructed a whole-gene nucleic acid vaccine from an isolated virulent Aleutian mink disease virus (ADV) strain (pcDNA3.1-ADV). Based on this whole-gene nucleic acid vaccine, we generated truncated mutant constructs by removing portions of the ADV VP2 gene using overlap extension polymerase chain reaction. pcDNA3.1-ADV-428 lacks nucleotides encoding VP2 amino acid residues 428-466, and pcDNA3.1-ADV-428-487 harbors additional deletion of nucleotides coding for VP2 amino acid residues 487-501. We also generated nucleic acid vaccines for the ADV NS1 gene, truncated ADV NS1 gene, ADV VS2 gene, and truncated ADV VS2 gene: pcDNA3.1-NS1, pcDNA3.1-NS1-D, pcDNA3.1-VP2, and pcDNA3.1-VP2-D, respectively. The immunogenicity of the seven DNA vaccines was confirmed by immunofluorescent evaluation. Sixty female minks were divided into 10 groups: seven groups were immunized with the DNA vaccines, one control group was injected with phosphate-buffered saline, one group was immunized with pcDNA3.1 empty vector, and one group was immunized with inactivated ADV-G virus. ADV antibody levels, percentage of CD8 cells in blood, and levels of γ-globulin and circulating immune complexes in the serum were evaluated longitudinally over 36 weeks after ADV challenge. Minks that were immunized with the pcDNA3.1-ADV-428-487 nucleic acid vaccine produced ADV antibodies. After ADV challenge, the minks immunized with pcDNA3.1-ADV-428-487 nucleic acid vaccine had lower γ-globulin content and lower CIC in serum compared to other immunization groups. Although the pcDNA3.1-ADV-428-487 nucleic acid vaccine did not demonstrate complete protection against ADV, it demonstrated marked efficacy and could potentially be used as a vaccine to prevent losses in mink populations due to ADV. Discovery of effective means to vaccinate mink against ADV will not only improve overall health of mink populations but will also reduce the economic impact of ADV.

摘要

水貂阿留申病(AD)是一种慢性病毒感染,可导致水貂出现自身免疫性疾病,并给水貂养殖带来重大经济负担。尽管AD带来了巨大挑战,但目前尚无有效的疫苗,仅实现了部分保护。我们从一株分离的强毒性水貂阿留申病病毒(ADV)毒株构建了全基因核酸疫苗(pcDNA3.1-ADV)。基于这种全基因核酸疫苗,我们使用重叠延伸聚合酶链反应去除ADV VP2基因的部分片段,生成了截短的突变体构建体。pcDNA3.1-ADV-428缺少编码VP2氨基酸残基428-466的核苷酸,而pcDNA3.1-ADV-428-487还额外缺失了编码VP2氨基酸残基487-501的核苷酸。我们还分别针对ADV NS1基因、截短的ADV NS1基因、ADV VS2基因和截短的ADV VS2基因生成了核酸疫苗:分别为pcDNA3.1-NS1、pcDNA3.1-NS1-D、pcDNA3.1-VP2和pcDNA3.1-VP2-D。通过免疫荧光评估证实了这七种DNA疫苗的免疫原性。60只雌性水貂被分为10组:7组用DNA疫苗免疫,1个对照组注射磷酸盐缓冲盐水,1组用pcDNA3.1空载体免疫,1组用灭活的ADV-G病毒免疫。在ADV攻击后36周内纵向评估ADV抗体水平、血液中CD8细胞百分比以及血清中γ-球蛋白和循环免疫复合物水平。用pcDNA3.1-ADV-428-487核酸疫苗免疫的水貂产生了ADV抗体。ADV攻击后,与其他免疫组相比,用pcDNA3.1-ADV-428-487核酸疫苗免疫的水貂血清中γ-球蛋白含量和CIC较低。尽管pcDNA3.1-ADV-428-487核酸疫苗未显示出对ADV的完全保护作用,但它显示出显著的效果,有可能用作预防水貂种群因ADV造成损失的疫苗。发现给水貂接种ADV疫苗的有效方法不仅将改善水貂种群的整体健康状况,还将减少ADV的经济影响。

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