Construction and Identification of Helper-Dependent Adenoviral Vector Encoding Hemagglutinin Protein of Swin-Origin Influenza Virus A H1N1_Journal of Biomedical Engineering

Authors：

ZHANGMei ¹ , JIANGYanze ² , CHENNianhua ³ , FUYuanhui ² , QIAOWei ^1,2 , WANGHe ² ,  HEJinsheng ^1,2

1. Department of Immunology, Anhui Medical University, Hefei 230032, China;
2. College of Life Sciences & Bioengineering, Beijing Jiaotong University, Beijing 100044, China;
3. Shandong Yantai No.2 Senior High School, Yantai 26400, China;

Corresponding author：

HEJinsheng, Email: jshhe@bjtu.edu.cn

Keywords：

helper-dependent adenoviral vector; swine-origin influenza virus A H1N1; hemagglutinin(HA)

DOI：

10.7507/1001-5515.20140031

Video：

Export PDF Favorites Scan Get Citation

Abstract Full text Figures/Tables Video References Cited by

In order to investigate immune protection against swine-origin influenza virus (S-OIV) A H1N1, the helper-dependent adenovirus vector (HDAd) system was exploited to construct recombinant HDAd encoding hemagglutinin (HA). The HA gene was synthesized and cloned to the HDAd backbone. Then, the HDAd/HA DNA molecules were transfected into 293Cre4 cells with calcium phosphate. The cells were infected by helper virus 16 hours after the transfection. The 293Cre4 cells were coinfected with HDAd/HA and the helper virus for large-scale preparation of HDAd/HA. The HDAd/HA was obtained and purified twice with CsCl density ultracentrifugation and observed morphologically under transmission electron microscope, and the expression of HA protein was analyzed with RT-PCR. Recombinant HDAd/HA expressing HA protein was successfully constructed which could pave the way for in vivo investigation on immunogenicity and efficacy against S-OIV A H1N1 infection.

Citation： ZHANGMei, JIANGYanze, CHENNianhua, FUYuanhui, QIAOWei, WANGHe, HEJinsheng. Construction and Identification of Helper-Dependent Adenoviral Vector Encoding Hemagglutinin Protein of Swin-Origin Influenza Virus A H1N1. Journal of Biomedical Engineering, 2014, 31(1): 157-160. doi: 10.7507/1001-5515.20140031 Copy

1.	COHEN J, ENSERINK M. As swine flu circles globe, scientists grapple with basic questions[J]. Science, 2009, 324(5927):572-573.
2.	PEIRIS J S, POON L L, GUAN Y. Emergence of a novel swine-origin influenza a virus (S-OIV) H1N1 virus in humans[J]. J Clin Virol, 2009, 45(3):169-173.
3.	张钫,郭骁才,程薇波,等.基于基因组序列特征的甲型H1N1流感病毒来源分析[J].生物医学工程学杂志,2010,27(4):868-874.
4.	WEAVER E A, NEHETE P N, BUCHL S S, et al. Comparison of replication-competent, first generation, and helper-dependent adenoviral vaccines[J]. PLoS One, 2009, 4(3):e5059.
5.	HARUI A, ROTH M D, KIERTSCHER S M, et al. Vaccination with helper-dependent adenovirus enhances the generation of transgene-specific CTL[J]. Gene Ther, 2004, 11(22):1617-1626.
6.	FU Y H, HE J S, ZHENG X X, et al. Intranasal vaccination with a helper-dependent adenoviral vector enhances transgene-specific immune responses in BALB/c mice[J]. Biochem Biophys Res Commun, 2010, 391(1):857-861.
7.	SHI C X, GRAHAM F L, HITT M M. A convenient plasmid system for construction of helper-dependent adenoviral vectors and its application for analysis of the breast-cancer-specific mammaglobin promoter[J]. J Gene Med, 2006, 8(4):442-451.
8.	郑娴娴,何金生,付远辉,等.辅助病毒依赖型腺病毒载体转基因的体外表达效率[J].生物工程学报,2010,26(8):1108-1115.
9.	杨兵,何金生,石长信,等.可表达人呼吸道合胞病毒融合蛋白辅助病毒依赖型腺病毒载体的构建与制备.微生物学报, 2007, 47(4):682-685.
10.	STEITZ J, BARLOW P G, HOSSAIN J, et al. A candidate H1N1 pandemic influenza vaccine elicits protective immunity in mice[J]. PLoS One, 2010, 5(5):e10492.
11.	GAO W, SOLOFF A C, LU X, et al. Protection of mice and poultry from lethal H5N1 avian influenza virus through adenovirus-based immunization[J]. J Virol, 2006, 80(4):1959-1964.
12.	严敏,孙茂盛,王文举,等.表达甲型H1N1流感病毒神经氨酸酶的重组腺病毒的构建及其免疫原性[J].中国生物制品学杂志,2011,24(11):1264-1268,1277.

1. COHEN J, ENSERINK M. As swine flu circles globe, scientists grapple with basic questions[J]. Science, 2009, 324(5927):572-573.
2. PEIRIS J S, POON L L, GUAN Y. Emergence of a novel swine-origin influenza a virus (S-OIV) H1N1 virus in humans[J]. J Clin Virol, 2009, 45(3):169-173.
3. 张钫,郭骁才,程薇波,等.基于基因组序列特征的甲型H1N1流感病毒来源分析[J].生物医学工程学杂志,2010,27(4):868-874.
4. WEAVER E A, NEHETE P N, BUCHL S S, et al. Comparison of replication-competent, first generation, and helper-dependent adenoviral vaccines[J]. PLoS One, 2009, 4(3):e5059.
5. HARUI A, ROTH M D, KIERTSCHER S M, et al. Vaccination with helper-dependent adenovirus enhances the generation of transgene-specific CTL[J]. Gene Ther, 2004, 11(22):1617-1626.
6. FU Y H, HE J S, ZHENG X X, et al. Intranasal vaccination with a helper-dependent adenoviral vector enhances transgene-specific immune responses in BALB/c mice[J]. Biochem Biophys Res Commun, 2010, 391(1):857-861.
7. SHI C X, GRAHAM F L, HITT M M. A convenient plasmid system for construction of helper-dependent adenoviral vectors and its application for analysis of the breast-cancer-specific mammaglobin promoter[J]. J Gene Med, 2006, 8(4):442-451.
8. 郑娴娴,何金生,付远辉,等.辅助病毒依赖型腺病毒载体转基因的体外表达效率[J].生物工程学报,2010,26(8):1108-1115.
9. 杨兵,何金生,石长信,等.可表达人呼吸道合胞病毒融合蛋白辅助病毒依赖型腺病毒载体的构建与制备.微生物学报, 2007, 47(4):682-685.
10. STEITZ J, BARLOW P G, HOSSAIN J, et al. A candidate H1N1 pandemic influenza vaccine elicits protective immunity in mice[J]. PLoS One, 2010, 5(5):e10492.
11. GAO W, SOLOFF A C, LU X, et al. Protection of mice and poultry from lethal H5N1 avian influenza virus through adenovirus-based immunization[J]. J Virol, 2006, 80(4):1959-1964.
12. 严敏,孙茂盛,王文举,等.表达甲型H1N1流感病毒神经氨酸酶的重组腺病毒的构建及其免疫原性[J].中国生物制品学杂志,2011,24(11):1264-1268,1277.

Journal of Biomedical Engineering

Construction and Identification of Helper-Dependent Adenoviral Vector Encoding Hemagglutinin Protein of Swin-Origin Influenza Virus A H1N1

Abstract Full text Figures/Tables Video References Cited by

Previous Article

Next Article

Format

Content