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表达猪圆环病毒2型Cap蛋白的重组伪狂犬病病毒的构建及其免疫原性

Generation and immunogenicity of a recombinant pseudorabies virus expressing cap protein of porcine circovirus type 2.

作者信息

Song Yunfeng, Jin Meilin, Zhang Songlin, Xu Xiaojuan, Xiao Shaobo, Cao Shengbo, Chen Huanchun

机构信息

Laboratory of Animal Virology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, PR China.

出版信息

Vet Microbiol. 2007 Jan 31;119(2-4):97-104. doi: 10.1016/j.vetmic.2006.08.026. Epub 2006 Sep 26.

Abstract

Porcine circovirus type 2 (PCV2) is associated with post-weaning multisystemic wasting syndrome (PMWS), which is an important economical disease affecting the pig industry worldwide. In order to develop an effective vaccine for PMWS, a recombinant pseudorabies virus (PRV) was generated and tested in piglets in this study. The PCV2 open reading frame 2 (ORF2) gene was inserted into pIECMV plasmid and co-transfected with PRV Tk-/gE-/LacZ+ genome into IBRS-2 cells to generate a recombinant Tk-/gE-/ORF2(+) virus. The expression of PCV2 ORF2 gene in the recombinant virus was confirmed by Western blotting and indirect immunofluorescence assay (IFA). Four-week-old piglets were immunized by the recombinant virus, and the immunogenicity of PRV Tk-/gE-/ORF2(+) was tested by PRV-enzyme-linked immunosorbent assay (ELISA), PRV neutralizing assay, ORF2-ELISA and ORF2 specific lymphocyte proliferation response. PRV Tk-/gE-/ORF2(+) elicited significant humoral immune responses to both PRV and PCV2, and the PCV2-specific lymphocyte proliferation response could be detected on day 49 of this experiment. These findings suggest that the recombinant PRV Tk-/gE-/ORF2(+) may be a potential vaccine against both PCV2 and PRV.

摘要

猪圆环病毒2型(PCV2)与断奶后多系统消耗综合征(PMWS)有关,PMWS是一种影响全球养猪业的重要经济疾病。为了开发一种针对PMWS的有效疫苗,本研究构建了一种重组伪狂犬病病毒(PRV)并在仔猪中进行了测试。将PCV2开放阅读框2(ORF2)基因插入pIECMV质粒,并与PRV Tk-/gE-/LacZ+基因组共转染到IBRS-2细胞中,以产生重组Tk-/gE-/ORF2(+)病毒。通过蛋白质免疫印迹法和间接免疫荧光试验(IFA)证实了PCV2 ORF2基因在重组病毒中的表达。用重组病毒对4周龄仔猪进行免疫,并通过PRV酶联免疫吸附测定(ELISA)、PRV中和试验、ORF2-ELISA和ORF2特异性淋巴细胞增殖反应来检测PRV Tk-/gE-/ORF2(+)的免疫原性。PRV Tk-/gE-/ORF2(+)引发了针对PRV和PCV2的显著体液免疫反应,并且在本实验的第49天可以检测到PCV2特异性淋巴细胞增殖反应。这些结果表明,重组PRV Tk-/gE-/ORF2(+)可能是一种针对PCV2和PRV的潜在疫苗。

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