Multivalent Virus-Like-Particle Vaccine Protects Against Classic and Variant Infectious Bursal Disease Viruses
Multivalent Virus-Like-Particle Vaccine Protects Against Classic and Variant Infectious Bursal Disease Viruses
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DOI：
10.1637/10312-080212-reg.1
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发表时间：
2013-03-01
期刊：
影响因子：
1.4
通讯作者：
Jackwood, Daral J.
中科院分区：
文献类型：
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作者：
Jackwood, Daral J.
利用逆转录-聚合酶链反应（RT-PCR）从命名为USA 08 MD 34 p的传染性法氏囊病病毒（IBDV）变异株和命名为Mo 195的IBDV经典株中产生编码pVP 2蛋白的核苷酸序列，并将其克隆到pGEM-T Easy载体中。还使用RT-PCR从USA 08 MD 34 p病毒基因组产生编码VP 3蛋白的核苷酸序列，并克隆到pGEM-T Easy载体中。将VP 3和pVP 2克隆插入pVL 1393杆状病毒转移载体中并测序，以确认它们对启动子的定向，并确保它们含有不间断的开放阅读框。通过转染Sf 9细胞构建重组杆状病毒。产生了三种重组杆状病毒，它们含有USA 08 MD 34 p-VP 3、USA 08 MD 34 pp-VP 2或Mo 195-pVP 2基因组序列。当USA 08 MD 34 p-VP 3杆状病毒与任一pVP 2构建体共接种到Sf 9细胞中时，使用透射电子显微镜观察到病毒样颗粒（VLP）。当USA 08 MD 34 p-pVP 2和Mo 195-pVP 2与USA 08 MD 34 p-VP 3共表达时，也观察到VLP。这些多价VLP含有经典和变体pVP 2分子。稳定性试验证明VLP在4 ℃和24 ℃下稳定8周。USA 08 MD 34 p、Mo 195和多价VLP用于接种鸡。它们诱导了通过酶联免疫吸附试验（ELISA）检测到的IBDV特异性抗体应答，并在体外检测到病毒中和抗体。用多价VLP接种的鸡被保护免于强毒变体IBDV毒株（V1）和强毒经典IBDV毒株（STC）。结果表明，多价VLP保持了变异病毒和经典病毒的抗原完整性，并有可能作为多价疫苗用于种鸡群疫苗接种计划。
Nucleotide sequences that encode the pVP2 proteins from a variant infectious bursal disease virus (IBDV) strain designated USA08MD34p and a classic IBDV strain designated Mo195 were produced with the use of reverse-transcriptase-polymerase chain reaction (RT-PCR) and cloned into a pGEM-T Easy vector. A nucleotide sequence that encodes the VP3 protein was also produced from the USA08MD34p viral genome with the use of RT-PCR and cloned into a pGEM-T Easy vector. The VP3 and pVP2 clones were inserted into the pVL1393 baculovirus transfer vector and sequenced to confirm their orientation to the promoter and to ensure they contained uninterrupted open reading frames. Recombinant baculoviruses were constructed by transfection in Sf9 cells. Three recombinant baculoviruses were produced and contained the USA08MD34p-VP3, USA08MD34pp-VP2, or Mo195-pVP2 genomic sequences. Virus-like particles (VLPs) were observed with the use of transmission electron microscopy when the USA08MD34p-VP3 baculovirus was co-inoculated into Sf9 cells with either of the pVP2 constructs. VLPs were also observed when the USA08MD34p-pVP2 and Mo195-pVP2 were coexpressed with USA08MD34p-VP3. These multivalent VLPs contained both classic and variant pVP2 molecules. Stability tests demonstrated the VLPs were stable at 4 and 24 C for 8 wk. The USA08MD34p, Mo195, and multivalent VLPs were used to vaccinate chickens. They induced an IBDV-specific antibody response that was detected by enzyme-linked immunosorbent assay (ELISA), and virus-neutralizing antibodies were detected in vitro. Chickens vaccinated with the multivalent VLPs were protected from a virulent variant IBDV strain (V1) and a virulent classic IBDV strain (STC). The results indicate the multivalent VLPs maintained the antigenic integrity of the variant and classic viruses and have the potential to serve as a multivalent vaccine for use in breeder-flock vaccination programs.