Efficient heterologous antigen gene delivery and expression by a replication-attenuated BoHV-4-based vaccine vector
Efficient heterologous antigen gene delivery and expression by a replication-attenuated BoHV-4-based vaccine vector
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DOI：
10.1016/j.vaccine.2013.06.052
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发表时间：
2013-08-20
期刊：
影响因子：
5.5
通讯作者：
Donofrio, Gaetano
中科院分区：
文献类型：
--
作者：
Capocefalo, Antonio;Mangia, Carlo;Donofrio, Gaetano
牛疱疹病毒4型（BoHV-4）是属于杆状病毒属的γ疱疹病毒，并且由于其生物学特性，已被提议作为用于兽用疫苗的疫苗载体。由于病毒载体相关风险是病毒载体应用的主要关注点，因此减毒是理想的特征。因此，人们致力于开发高度减毒的病毒载体。BoHV-4天然地表现出有限的致病性，并且通过破坏编码1.7-kb多聚腺苷酸化RNA（L1.7）的晚期基因获得了在复制方面的进一步减毒。通过从克隆为细菌人工染色体（BAC）的BoHV-4分离物（BoHV-4-A）的基因组同源重组产生L1.7缺失突变体BoHV-4（BoHV-4-A-KanaGaIK Delta L1.7）及其回复突变体（BoHV-4-A-Rev）。当与BoHV-4-A、BowHV-4-A-Rev和BoHV-4-A-KanaGalK Delta TK（其中KanaGalK选择性标记插入胸苷激酶开放阅读框的重组对照病毒）相比时，BoHV-4-A-KanaGalK Delta L1.7在重构感染性病毒的能力、病毒复制和噬斑大小方面显示出减弱。通过用水泡性口炎病毒糖蛋白（VSVg）表达盒替换KanaGalK盒以产生BoHV-4A-EF 1 α VSVg Δ L1.7来研究BoHV-4-A-KanaGalK Δ L1.7递送和表达异源抗原的能力。BoHV-4-A-EF 1 alpha NSVg Delta L1.7感染的细胞稳健地表达VSVg，因此证实了由L1.7破坏引起的复制缺陷不会阻止异源基因递送和表达。尽管需要进一步的工作来鉴定BoHV-4 L1.7基因的特异性功能，但L1.7基因可能代表了用于整合异源抗原表达盒的理想靶向位点，从而导致病毒载体的减毒。(C)2013爱思唯尔有限公司保留所有权利。
Bovine Herpesvirus 4 (BoHV-4) is a gammaherpesvirus belonging to the Rhadinovirus genus and due to its biological characteristics has been proposed as a vaccine vector for veterinary vaccines. Because viral vector-associated risk is a major concern for viral vector applications, attenuation is a desirable feature. Therefore, efforts are directed toward the development of highly attenuated viral vectors. BoHV-4 naturally exhibits limited pathogenicity and a further attenuation, in terms of replication, was obtained by disrupting the late gene encoding the 1.7-kb polyadenylated RNA (L1.7). An L1.7 deleted mutant BoHV-4 (BoHV-4-A-KanaGaIK Delta L1.7), as well as its revertant (BoHV-4-A-Rev), was generated by homologous recombination from the genome of a BoHV-4 isolate (BoHV-4-A) cloned as a bacterial artificial chromosome (BAC). BoHV-4-A-KanaGaIK Delta L1.7 showed attenuation in terms of competence to reconstitute infectious virus, viral replication, and plaque size when compared to BoHV-4-A, BowHV-4-A-Rev, and BoHV-4-A-KanaGalK Delta TK, a recombinant control virus where the KanaGalK selectable marker was inserted into the thymidine kinase open reading frame. The capability of BoHV-4-A-KanaGalK Delta L1.7 to deliver and express a heterologous antigen was investigated by replacing the KanaGalK cassette with a vesicular stomatitis virus glycoprotein (VSVg) expression cassette to generate BoHV-4A-EF1 alpha VSVg Delta L1.7. BoHV-4-A-EF1 alpha NSVg Delta L1.7 infected cells robustly expressed VSVg, thus confirming that the replication deficiency resulting from L1.7 disruption did not prevent heterologous gene delivery and expression. Although further work is needed to identify the specific function of the BoHV-4 L1.7 gene, the L1.7 gene may represent an ideal targeting locus for the integration of a heterologous antigen expression cassette, resulting in attenuation of the viral vector. (C) 2013 Elsevier Ltd. All rights reserved.