Generation and Immunogenicity of a Recombinant Adenovirus Co-Expressing the E2 Protein of Classical Swine Fever Virus and the GP5 Protein of Porcine Reproduction and Respiratory Syndrome Virus

Agricultural Sciences in China Pub Date : 2011-11-01 DOI:10.1016/S1671-2927(11)60178-8

Hong-yu LI, Yuan SUN, Xing-juan ZHANG, Tian-ming CHANG, Xiang-peng WANG, Fan HE, Jun-hua HUANG, Hua-ji QIU

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Abstract

Classical swine fever (CSF) and porcine reproduction and respiratory syndrome (PRRS) are both economically important, highly contagious diseases of swine worldwide. To develop an effective vaccine to control these two diseases, we constructed a recombinant adenovirus rAdV-GP52AE2, using a replication-defective human adenovirus serotype 5 as a delivery vector, to co-express the GP5 protein of highly pathogenic porcine reproduction and respiratory syndrome virus (PRRSV) and the E2 protein of classical swine fever virus (CSFV). Foot-and-mouth disease virus (FMDV) 2A peptide was used as a linker between the GP5 and E2 proteins to allow automatic self-cleavage of the polyprotein. The GP5 and E2 genes were expressed as demonstrated by immunofluorescence assay and Western blotting. Immunization of mice resulted in a CSFV-neutralizing antibody titer of 1:128 and a PRRSV-neutralizing antibody titer of 1:16. The lymphoproliferative responses were detected by Cell Counting Kit-8 assay and the stimulation index of CFSV-specific and PRRSV-specific lymphocytes in the rAdV-GP52AE2 group was significantly higher than that in the negative control group. The results show that rAdV-GP52AE2 can induce both effective humoral and cell-mediated immune responses in mice. The protective efficacy of the recombinant virus against CSF was evaluated in immunized rabbits, which were protected from fever induced by challenge with C-strain. Our study provides supporting evidence for the use of FMDV 2A to develop a bivalent genetically-engineered vaccine.

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共表达猪瘟病毒E2蛋白和猪繁殖与呼吸综合征病毒GP5蛋白的重组腺病毒的生成及免疫原性

摘要猪瘟(CSF)和猪繁殖与呼吸综合征(PRRS)是世界范围内具有重要经济意义和高度传染性的猪传染病。为了研制有效的疫苗，我们构建了重组腺病毒rAdV-GP52AE2，以复制缺陷型5型人腺病毒为载体，共表达高致病性猪繁殖与呼吸综合征病毒(PRRSV)的GP5蛋白和猪瘟病毒(CSFV)的E2蛋白。利用口蹄疫病毒(FMDV) 2A肽作为GP5和E2蛋白之间的连接物，实现多蛋白的自动自裂。免疫荧光法和免疫印迹法证实GP5和E2基因均有表达。小鼠免疫后的猪瘟中和抗体效价为1:128,prrsv中和抗体效价为1:16。细胞计数试剂盒-8检测淋巴细胞增殖反应，rAdV-GP52AE2组cfv特异性和prrsv特异性淋巴细胞刺激指数显著高于阴性对照组。结果表明，rAdV-GP52AE2可诱导小鼠有效的体液和细胞介导的免疫应答。用c -菌株攻毒的免疫家兔，评价了重组病毒对脑脊液的保护作用。我们的研究为使用FMDV 2A开发一种二价基因工程疫苗提供了支持证据。

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来源期刊

Agricultural Sciences in China AGRICULTURE, MULTIDISCIPLINARY-

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审稿时长

3.2 months