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

机译:表达2型猪圆环病毒帽蛋白的重组伪狂犬病毒的产生和免疫原性

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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.
机译:猪圆环病毒2型(PCV2)与断奶后多系统消耗综合症(PMWS)相关,这是一种影响全球养猪业的重要经济疾病。为了开发有效的PMWS疫苗,本研究中产生了重组伪狂犬病病毒(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IBRS-2细胞,以产生重组Tk- / gE- / ORF2(+)病毒。 Western blotting和间接免疫荧光法(IFA)证实了重组病毒中PCV2 ORF2基因的表达。用重组病毒对4周龄的仔猪进行免疫,并通过PRV酶联免疫吸附试验(ELISA),PRV中和试验,ORF2-ELISA和PRV检测PRV Tk- / gE- / ORF2(+)的免疫原性。 ORF2特异性淋巴细胞增殖反应。 PRV Tk- / gE- / ORF2(+)引起对PRV和PCV2的显着体液免疫反应,并且在该实验的第49天可以检测到PCV2特异性淋巴细胞增殖反应。这些发现表明重组PRV Tk- / gE- / ORF2(+)可能是针对PCV2和PRV的潜在疫苗。

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