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对于作为鸡用疫苗的杆状病毒，白斑综合征病毒ie1启动子在表达来自流感病毒的H5血凝素方面比巨细胞病毒启动子更有效。

WSSV ie1 promoter is more efficient than CMV promoter to express H5 hemagglutinin from influenza virus in baculovirus as a chicken vaccine.

作者信息

He Fang, Ho Yuenfern, Yu Li, Kwang Jimmy

机构信息

Animal Health Biotechnology, Temasek Life Sciences Laboratory, National University of Singapore, Singapore.

出版信息

BMC Microbiol. 2008 Dec 31;8:238. doi: 10.1186/1471-2180-8-238.

DOI:10.1186/1471-2180-8-238
PMID:19116038
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC2631607/
Abstract

BACKGROUND

The worldwide outbreak of influenza A (H5N1) viruses among poultry species and humans highlighted the need to develop efficacious and safe vaccines based on efficient and scaleable production.

RESULTS

White spot syndrome virus (WSSV) immediate-early promoter one (ie1) was shown to be a stronger promoter for gene expression in insect cells compared with Cytomegalovirus immediate-early (CMV) promoter in luciferase assays. In an attempt to improve expression efficiency, a recombinant baculovirus was constructed expressing hemagglutinin (HA) of H5N1 influenza virus under the control of WSSV ie1 promoter. HA expression in SF9 cells increased significantly with baculovirus under WSSV ie1 promoter, compared with CMV promoter based on HA contents and hemagglutination activity. Further, immunization with baculovirus under WSSV ie1 promoter in chickens elicited higher level anti-HA antibodies compared to CMV promoter, as indicated in hemagglutination inhibition, virus neutralization and enzyme-linked immunosorbent assays. By immunohistochemistry, strong HA antigen expression was observed in different chicken organs with vaccination of WSSV ie1 promoter controlled baculovirus, confirming higher efficiency in HA expression by WSSV ie1 promoter.

CONCLUSION

The production of H5 HA by baculovirus was enhanced with WSSV ie1 promoter, especially compared with CMV promoter. This contributed to effective elicitation of HA-specific antibody in vaccinated chickens. This study provides an alternative choice for baculovirus based vaccine production.

摘要

背景

甲型流感病毒（H5N1）在全球家禽和人类中的爆发凸显了基于高效且可规模化生产来开发有效且安全疫苗的必要性。

结果

在荧光素酶测定中，与巨细胞病毒立即早期（CMV）启动子相比，白斑综合征病毒（WSSV）立即早期启动子1（ie1）在昆虫细胞中是一种更强的基因表达启动子。为提高表达效率，构建了一种重组杆状病毒，其在WSSV ie1启动子的控制下表达H5N1流感病毒的血凝素（HA）。基于HA含量和血凝活性，与CMV启动子相比，在WSSV ie1启动子控制下的杆状病毒使SF9细胞中的HA表达显著增加。此外，如血凝抑制、病毒中和及酶联免疫吸附测定所示，用WSSV ie1启动子控制的杆状病毒免疫鸡后，引发的抗HA抗体水平高于CMV启动子。通过免疫组织化学，在用WSSV ie1启动子控制的杆状病毒疫苗接种的不同鸡器官中观察到强烈的HA抗原表达，证实WSSV ie1启动子在HA表达方面具有更高效率。

结论

WSSV ie1启动子增强了杆状病毒对H5 HA的生产，特别是与CMV启动子相比。这有助于在接种疫苗的鸡中有效引发HA特异性抗体。本研究为基于杆状病毒的疫苗生产提供了一种替代选择。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/07e8/2631607/26c95f975c04/1471-2180-8-238-4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/07e8/2631607/0ef755689aa3/1471-2180-8-238-1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/07e8/2631607/252d68193b64/1471-2180-8-238-2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/07e8/2631607/89c3155f6b33/1471-2180-8-238-3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/07e8/2631607/26c95f975c04/1471-2180-8-238-4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/07e8/2631607/0ef755689aa3/1471-2180-8-238-1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/07e8/2631607/252d68193b64/1471-2180-8-238-2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/07e8/2631607/89c3155f6b33/1471-2180-8-238-3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/07e8/2631607/26c95f975c04/1471-2180-8-238-4.jpg

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