A rapid strategy for constructing novel simian adenovirus vectors with high viral titer and expressing highly antigenic proteins applicable for vaccine development
A rapid strategy for constructing novel simian adenovirus vectors with high viral titer and expressing highly antigenic proteins applicable for vaccine development
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构建高病毒滴度的新型猿猴腺病毒载体并表达适用于疫苗开发的高抗原蛋白的快速策略
DOI：
10.1016/j.virusres.2019.05.008
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发表时间：
2019-07-15
期刊：
影响因子：
5
通讯作者：
Li, Tingting
中科院分区：
文献类型：
--
作者：
Luo, Shengxue;Zhang, Panli;Li, Tingting
腺病毒载体已被广泛用于开发感染性疾病疫苗。然而，源于主要腺病毒血清型5株的人腺病毒载体的挑战限制了其在接受者中广泛存在的中和抗体的有用性。为了克服这一障碍，我们在人类或灵长类动物中产生了一种特设腺病毒载体。在此，通过吉布森组装，构建嵌合猿猴腺病毒载体Sad 23，其包括缺失全长猿猴腺病毒血清型23基因组（SAdV 23）的E1和E3区。为了提高Sad 23病毒的增殖效率，将E4区开放阅读框6（orf 6）替换为人5型腺病毒（Ad 5）的相应元件，命名为Sad 23 L。该载体的克隆过程仅需一周时间，重组腺病毒在HEK 293细胞中以高滴度包装。为了验证这种新型腺病毒载体递送外源基因的能力，使用寨卡病毒（ZIKV）prM-E基因作为抗原表达的靶基因。将重组腺病毒Sad 23 L-prM-E、Sad 23-prM-E和Ad 5-prM-E肌内接种到Ad 5-eGFP未预暴露或预暴露的小鼠中，并比较载体之间对ZIKV prM-E的免疫应答。Sad 23 L-prM-E在Ad 5预暴露小鼠中诱导了相当稳健的免疫应答并保持了免疫原性，这表明Ad 5预先存在的免疫力不影响Sad 23 L-prM-E免疫。这些初步结果表明，所提出的快速策略是有效的，在构建一个新的腺病毒载体平台（Sad 23 L）可用于开发的人疫苗。
Adenoviral vectors have been widely used for the development of infectious disease vaccines. However, the challenge of human adenoviral vector rooted from the predominant adenovirus serotype 5 strain limiting its usefulness by the widespread pre-existing neutralizing antibodies in recipients. To circumvent this obstacle, we generated an ad-hoc adenovirus vector in human or primates. Here, a chimeric simian adenoviral vector Sad23 was constructed consisting in deleting of El and E3 regions of the full-length simian adenovirus serotype 23 genome (SAdV23) by Gibson assembly. To improve Sad23 virus propagating efficiency, the E4 region open reading frame 6 (orf6) was replaced by the corresponding element of human adenovirus type 5 (Ad5), designated Sad23L. The procedure for cloning this novel vector took a single week, and recombinant adenovirus was packaged with high titer in HEK293 cells. To verify the ability of this novel adenoviral vector to deliver foreign genes, Zika virus (ZIKV) prM-E genes were used as target genes for antigen expression. Recombinant adeno-viruses Sad23L-prM-E, Sad23-prM-E and Ad5-prM-E were intramuscularly inoculated into Ad5-eGFP none pre exposed or pre-exposed mice, and the immune response to ZIKV prM-E was compared between vectors. Sad23L-prM-E induced a fairly robust immune response and maintained immunogenicity in Ad5 pre-exposed mice, which suggested that Ad5 pre-existing immunity did not affect Sad23L-prM-E immunization. These preliminary results suggest that the proposed rapid strategy was effective in constructing a new adenoviral vector platform (Sad23 L) usable for the development of human vaccines.