Construction and Characterization of a Novel Recombinant Attenuated and Replication-Deficient Candidate Human Adenovirus Type 3 Vaccine: &quot;Adenovirus Vaccine Within an Adenovirus Vector&quot;

Yuqian Yan; Shuping Jing; Liqiang Feng; Jing Zhang; Zhiwei Zeng; Min Li; Shan Zhao; Junxian Ou; Wendong Lan; Wenyi Guan; Xiaowei Wu; Jianguo Wu; Donald Seto; Qiwei Zhang

doi:10.1007/s12250-020-00234-1

June 2021

Yuqian Yan, Shuping Jing, Liqiang Feng, Jing Zhang, Zhiwei Zeng, Min Li, Shan Zhao, Junxian Ou, Wendong Lan, Wenyi Guan, Xiaowei Wu, Jianguo Wu, Donald Seto and Qiwei Zhang. Construction and Characterization of a Novel Recombinant Attenuated and Replication-Deficient Candidate Human Adenovirus Type 3 Vaccine: 'Adenovirus Vaccine Within an Adenovirus Vector'[J]. Virologica Sinica, 2021, 36(3): 354-364. doi: 10.1007/s12250-020-00234-1

Citation: Yuqian Yan, Shuping Jing, Liqiang Feng, Jing Zhang, Zhiwei Zeng, Min Li, Shan Zhao, Junxian Ou, Wendong Lan, Wenyi Guan, Xiaowei Wu, Jianguo Wu, Donald Seto, Qiwei Zhang. Construction and Characterization of a Novel Recombinant Attenuated and Replication-Deficient Candidate Human Adenovirus Type 3 Vaccine: "Adenovirus Vaccine Within an Adenovirus Vector" .VIROLOGICA SINICA, 2021, 36(3) : 354-364. http://dx.doi.org/10.1007/s12250-020-00234-1

一种新型的人3型腺病毒复制缺陷型重组减毒疫苗候选株的构建及特性：腺病毒载体用于腺病毒疫苗

严雨茜 ^1, ,
京舒娉 ^1,2, ,
冯立强 ³ ,
张静 ⁴ ,
曾志伟 ¹ ,
李敏 ¹ ,
赵珊 ¹ ,
欧俊贤 ¹ ,
蓝文东 ¹ ,
管文艺 ¹ ,
吴晓薇 ¹ ,
吴建国 ⁴ ,
SetoDonald ⁵ ,
张其威 ^1,4,,

1.
南方医科大学
2.
珠海市疾病预防控制中心
3.
中国科学院广州生物医药与健康研究院
4.
暨南大学
5.
美国乔治梅森大学

通讯作者： 张其威, zhangqw@smu.edu.cn, ORCID: http://orcid.org/0000-0002-2770-111X
收稿日期： 2020-02-22
录用日期： 2020-04-13
出版日期： 2020-05-26

摘要

人腺病毒（HAdVs）传染性强，引起大量的急性呼吸系统疾病（ARDs）病例，具有较高的发病率和一定的致死率。在亚洲、欧洲和美洲，人腺病毒3型（HAdV-3）均是导致ARD暴发最常见的腺病毒型别。然而，目前还没有在普通人群中批准使用的疫苗。六邻体（hexon）蛋白含有腺病毒主要的中和表位，具有强大和持久的免疫原性。在本研究中，以基因治疗和疫苗载体研究中E1区缺失的复制缺陷型HAdV-5商品化载体为基础，我们构建了一种新型的重组减毒的人3型腺病毒疫苗候选株。利用细菌内同源重组的方法，将整个HAdV-3的六邻体基因整合到载体的E1区。在AD293细胞中成功包装出重组病毒，并表达HAdV-3的六邻体蛋白，随后用逆转录PCR、免疫印迹、间接免疫荧光和电镜观察等方法鉴定验证正确。这种潜在的疫苗候选株与野生型HAdV-3株在互补细胞系AD293内具有相似的复制能力。然而，更重要的是，疫苗候选株在AD293细胞中经20代以上的连续传代后，仍不能在人肺腺癌A549细胞中复制，持续保持其复制缺陷的特性，这表明该疫苗具有非常高的安全性。通过滴鼻或肌肉注射途径，该候选疫苗株免疫的小鼠均能产生较高滴度的针对HAdV-3的中和性抗体。因此，该重组、减毒且安全的腺病毒疫苗是一种很有发展前景的HAdV-3疫苗候选株。这种使用临床批准的复制缺陷型HAdV-5载体用于腺病毒自身疫苗研究的策略，为研发腺病毒疫苗提供了一种新的思路和方法，今后可用于引起呼吸系统疾病以及其它相关疾病的所有型别腺病毒的通用疫苗的构建和研发。
- 腺病毒疫苗
- , 人3型腺病毒（HAdV-3）
- , 复制缺陷型腺病毒载体
- , BALB/c小鼠免疫
- , 重组

Construction and Characterization of a Novel Recombinant Attenuated and Replication-Deficient Candidate Human Adenovirus Type 3 Vaccine: "Adenovirus Vaccine Within an Adenovirus Vector"

Yuqian Yan ^1, ,
Shuping Jing ^1,2, ,
Liqiang Feng ³ ,
Jing Zhang ⁴ ,
Zhiwei Zeng ¹ ,
Min Li ¹ ,
Shan Zhao ¹ ,
Junxian Ou ¹ ,
Wendong Lan ¹ ,
Wenyi Guan ¹ ,
Xiaowei Wu ¹ ,
Jianguo Wu ⁴ ,
Donald Seto ⁵ ,
Qiwei Zhang ^1,4,,

1.
Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical University, Guangzhou 510515, China
2.
Microbiological Laboratory, Zhuhai Center for Disease Control and Prevention, Zhuhai 519000, China
3.
Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China
4.
Guangdong Provincial Key Laboratory of Virology, Institute of Medical Microbiology, Jinan University, Guangzhou 510632, China
5.
Bioinformatics and Computational Biology Program, School of Systems Biology, George Mason University, Manassas, VA 20110, USA

Corresponding author: Qiwei Zhang, zhangqw@smu.edu.cn
ORCID: http://orcid.org/0000-0002-2770-111X
Received Date: 22 February 2020
Accepted Date: 13 April 2020
Published Date: 26 May 2020

Abstract

Human adenoviruses (HAdVs) are highly contagious and result in large number of acute respiratory disease (ARD) cases with severe morbidity and mortality. Human adenovirus type 3 (HAdV-3) is the most common type that causes ARD outbreaks in Asia, Europe, and the Americas. However, there is currently no vaccine approved for its general use. The hexon protein contains the main neutralizing epitopes, provoking strong and lasting immunogenicity. In this study, a novel recombinant and attenuated adenovirus vaccine candidate against HAdV-3 was constructed based on a commercially-available replication-defective HAdV-5 gene therapy and vaccine vector. The entire HAdV-3 hexon gene was integrated into the E1 region of the vector by homologous recombination using a bacterial system. The resultant recombinants expressing the HAdV-3 hexon protein were rescued in AD293 cells, identified and characterized by RT-PCR, Western blots, indirect immunofluorescence, and electron microscopy. This potential vaccine candidate had a similar replicative efficacy as the wild-type HAdV-3 strain. However, and importantly, the vaccine strain had been rendered replication-defective and was incapable of replication in A549 cells after more than twenty-generation passages in AD293 cells. This represents a significant safety feature. The mice immunized both intranasally and intramuscularly by this vaccine candidate raised significant neutralizing antibodies against HAdV-3. Therefore, this recombinant, attenuated, and safe adenovirus vaccine is a promising HAdV-3 vaccine candidate. The strategy of using a clinically approved and replication-defective HAdV-5 vector provides a novel approach to develop universal adenovirus vaccine candidates against all the other types of adenoviruses causing ARDs and perhaps other adenovirus-associated diseases.
- Adenovirus vaccine
- , Human adenovirus type 3 (HAdV-3)
- , Replication-deficient adenovirus vector
- , Immunity in BALB/c mice
- , Recombination

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