嵌合猪瘟病毒基因Ⅱ型E2抗原区的重组C株病毒 构建及其生物学特性

doi:10.3864/j.issn.0578-1752.2013.01.021

摘要/Abstract

摘要： 【目的】目前使用的基因Ⅰ型猪瘟兔化弱毒C株疫苗是公认的安全、有效疫苗，但近年来基因Ⅱ群的猪瘟流行毒株比例增高，因此对猪瘟(CSF)的有效防控提出了更高要求。本研究旨在通过反向遗传学技术构建适应CSFV基因Ⅱ型的新型C株疫苗，为标记疫苗的开发奠定基础。【方法】在构建的C株感染性克隆基础上，拯救嵌合CSFV基因Ⅱ型HZ1-08毒株E2抗原区的重组C株病毒RecCHZE2，并分析其致病性和免疫原性。【结果】重组嵌合病毒RecCHZE2接种家兔后出现典型的定型热，兔脾脏肿大；重组嵌合病毒RecCHZE2接种猪只后没有发热和明显的白细胞减少，在接种后第12天检测到一过性的病毒血症；对接种猪血清检测结果表明，针对2型毒株的相应抗体在接种后的20 d达到高峰；接种猪血清针对C株和HZ1-08毒株的中和抗体效价相当，但C株免疫猪血清对流行毒株的中和抗体水平降低。【结论】重组嵌合病毒RecCHZE2不仅保留C株病毒弱毒的特性，且可提高针对Ⅱ型流行毒株的抗体水平。

关键词: 猪瘟病毒 , 重组嵌合C株病毒 , 致病性 , 免疫原性

Abstract: 【Objective】 The classical swine fever (CSF) C-strain vaccine has proven to be safe and highly efficacious. However, classical swine fever virus (CSFV) genotype Ⅱisolates became dominant in recent years, making it more challenging to bring the disease into total control. This study was attempted to engineer, by reverse genetic approach, a novel C-strain-based vaccine candidate that matches molecular variations of field isolates, and to characterize its biological features. Those results could provide foundations for further development of a marker vaccine for differential diagnostic purpose.【Method】An infectious recombinant CSF virus was generated by exchanging the 5’-end 870-bp of E2 gene in C-strain with the same fragment from group 2 strain HZ1-08. The resulting recombinant virus RecCHZE2 was rescued and its pathogenicity and immunogenicity were characterized in rabbits and pigs. 【Result】 Rabbits inoculated with RecCHZE2 exhibited typical temperature profiles similar to the C-strain with spleen enlargement. Pigs inoculated with the recombinant chimeric virus did not show fever and significant decline of the number of white blood cells. Mild transient viremia was detected at day 12 post-inoculation. The antibody responses specific to the field strain HZ1-08 reached the peak 20 days post-inoculation. The neutralizing antibody titers from pigs immunized with RecCHZE2 were similar to both HZ1-08 and C strains while the titer from pigs immunized with RecC were lower for group 2 strain HZ1-08.【Conclusion】The recombinant virus RecCHZE2 remains avirulent but differs in immunogenic properties from the C-strain.

Key words: classical swine fever virus , chimeric C-strain virus , pathogenicity , immunogenicity

童超, 陈宁, 廖迅, 袁雪梅, 李肖梁, 方维焕. 嵌合猪瘟病毒基因Ⅱ型E2抗原区的重组C株病毒构建及其生物学特性[J]. 中国农业科学, 2013, 46(1): 179-186.

TONG Chao, CHEN Ning, LIAO Xun, YUAN Xue-Mei, LI Xiao-Liang, FANG Wei-Huan. Construction of a Chimeric Classical Swine Fever Virus C-strain Containing E2 of Group Ⅱ Isolate Remains and Its Biological Characteristics[J]. Scientia Agricultura Sinica, 2013, 46(1): 179-186.

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