羊口疮与绵羊痘二联重组DNA疫苗的构建及其免疫应答  

Construction and immune response of double recombinant DNA vaccine for contagious ecthyma and sheep pox

作　　者：王妍 文美龄 王仁铃 王蓬琨 关继羽 宋德光 陆慧君 WANG Yan;WEN Meiling;WANG Renling;WANG Pengkun;GUAN Jiyu;SONG Deguang;LU Huijun(College of Veterinary Medicine/Institute of Zoonosis,Jilin University,Changchun 130062,China)

机构地区：[1]吉林大学动物医学学院人兽共患病研究所,吉林长春130062

出　　处：《中国兽医学报》2023年第2期307-314,共8页Chinese Journal of Veterinary Science

基　　金：吉林省科技发展计划(重点研发项目)资助项目(20200402052NC)。

摘　　要：羊口疮(Orf)和绵羊痘(SPP)是发生于羊群的高度接触性传染病,两者在临床上经常呈现混合感染,给养羊业带来严重的经济损失,因此,开发安全有效、可同时抵抗2种疫病的二联疫苗具有重要临床意义。本研究将羊口疮病毒(ORFV)的B2L基因及绵羊痘病毒(SPPV)的P32基因以P2A肽序列串联,获得pcDNA3.1-B2L-P2A-P32质粒,作为候选重组DNA疫苗。将质粒转染细胞,检测抗原蛋白的胞内表达及切割;将其免疫BALB/c小鼠,以间接ELISA法检测特异性抗体、中和抗体效价和细胞因子的分泌水平。结果表明:该重组DNA疫苗能在真核细胞中独立表达B2L及P32抗原蛋白,同时能诱导机体产生特异性免疫应答,具有较好的免疫效果。该研究为在羊群中开展更广泛的临床研究提供了试验依据,有望成为候选疫苗。Contagious ecthyma(Orf)and sheep pox(SPP)are highly contagious infectious diseases that frequently occur in sheep.Sheep were often mixed infected in clinic,which brings great economic losses to the sheep farming industry.It is of great significance to develop a safe and effective dual vaccine with simultaneous resistance to both of these two diseases.In this study,B2L gene from orf virus(ORFV)and P32 gene from sheep pox virus(SPPV)were linked with P2A sequence,which were cloned into a eukaryotic expression vector.The recombinant plasmid pcDNA3.1-B2L-P2A-P32 was obtained as a recombinant DNA vaccine candidate.The recombinant plasmid was transfected into cells to detect the intracellular expression and cleavage of these two antigenic proteins.BALB/c mice were immunized,and specific antibodies,neutralizing antibodies and cytokine secretion level were detected by indirect ELISA.The results showed that the recombinant DNA vaccine could express B2L and P32 antigenproteins in eukaryotic cells,and could induce specific immune responses in the body.This work provides an experimental basis for more extensive clinical research in sheep,and can be used as a vaccine candidate for in-depth research.

关 键 词：羊口疮病毒 绵羊痘病毒 重组二联DNA疫苗 

分 类 号：S852.65[农业科学—基础兽医学]