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杆状病毒载体疫苗诱导流感病毒血凝素免疫应答的特性研究。

Characterization of the immune responses elicited by baculovirus-based vector vaccines against influenza virus hemagglutinin.

机构信息

Department of Microbiology and Microbial Engineering, School of Life Sciences, Fudan University, Shanghai, China.

出版信息

Acta Pharmacol Sin. 2012 Jun;33(6):783-90. doi: 10.1038/aps.2012.23. Epub 2012 May 7.

DOI:10.1038/aps.2012.23
PMID:22562016
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4010374/
Abstract

AIM

To compare the specific immune responses elicited by different baculovirus vectors in immunized mice.

METHODS

We constructed and characterized two recombinant baculoviruses carrying the expression cassette for the H5N1 influenza virus hemagglutinin (HA) gene driven by either an insect cell promoter (vAc-HA) or a dual-promoter active both in insect and mammalian cells (vAc-HA-DUAL). Virus without the HA gene (vAc-EGFP) was used as a control. These viruses were used to immunize mice subcutaneously and intraperitoneally. The production of total and specific antibodies was determined by ELISA and competitive ELISA. Cytokine production by the spleen cells of immunized mice was studied using the ELISPOT assay.

RESULTS

Both the vAc-HA and vAc-HA-DUAL vectors expressed HA proteins in insect Sf9 cells, and HA antigen was present in progeny virions. The vAc-HA-DUAL vector also mediated HA expression in virus-transduced mammalian cell lines (BHK and A547). Both vAc-HA and vAc-HA-DUAL exhibited higher transduction efficiencies than vAc-EGFP in mammalian cells, as shown by the expression of the reporter gene egfp. Additionally, both vAc-HA and vAc-HA-DUAL induced high levels of HA-specific antibody production in immunized mice; vAc-HA-DUAL was more efficient in inducing IFN-γ and IL-2 upon stimulation with specific antigen, whereas vAc-HA was more efficient in inducing IL-4 and IL-6.

CONCLUSION

Baculovirus vectors elicited efficient, specific immune responses in immunized mice. The vector displaying the HA antigen on the virion surface (vAc-HA) elicited a Th2-biased immune response, whereas the vector displaying HA and mediating HA expression in the cell (vAc-HA-DUAL) elicited a Th1-biased immune response.

摘要

目的

比较不同杆状病毒载体在免疫小鼠中诱导的特异性免疫应答。

方法

我们构建并鉴定了两个携带 H5N1 流感病毒血凝素（HA）基因表达盒的重组杆状病毒，该基因由昆虫细胞启动子（vAc-HA）或在昆虫和哺乳动物细胞中均活跃的双启动子（vAc-HA-DUAL）驱动。不含 HA 基因的病毒（vAc-EGFP）用作对照。这些病毒被用于皮下和腹腔内免疫小鼠。通过 ELISA 和竞争 ELISA 测定总抗体和特异性抗体的产生。通过 ELISPOT 测定研究免疫小鼠脾细胞的细胞因子产生。

结果

vAc-HA 和 vAc-HA-DUAL 载体均在昆虫 Sf9 细胞中表达 HA 蛋白，并且 HA 抗原存在于子代病毒粒子中。vAc-HA-DUAL 载体还介导了转导的哺乳动物细胞系（BHK 和 A547）中的 HA 表达。vAc-HA 和 vAc-HA-DUAL 载体在哺乳动物细胞中的转导效率均高于 vAc-EGFP，如报告基因 egfp 的表达所示。此外，vAc-HA 和 vAc-HA-DUAL 均可在免疫小鼠中诱导高水平的 HA 特异性抗体产生；vAc-HA-DUAL 在刺激特异性抗原时更有效地诱导 IFN-γ 和 IL-2 的产生，而 vAc-HA 更有效地诱导 IL-4 和 IL-6 的产生。

结论

杆状病毒载体在免疫小鼠中引起有效的特异性免疫应答。在病毒表面显示 HA 抗原的载体（vAc-HA）引起 Th2 偏向的免疫应答，而在细胞中显示 HA 并介导 HA 表达的载体（vAc-HA-DUAL）引起 Th1 偏向的免疫应答。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e529/4010374/8d359832ea2f/aps201223f8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e529/4010374/3c85039b3cfa/aps201223f1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e529/4010374/0dc19f16df86/aps201223f2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e529/4010374/41652ffb50a0/aps201223f3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e529/4010374/eb2bf890387e/aps201223f4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e529/4010374/cc5c79908df8/aps201223f5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e529/4010374/d996dc85ab54/aps201223f6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e529/4010374/bfc1e40979bc/aps201223f7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e529/4010374/8d359832ea2f/aps201223f8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e529/4010374/3c85039b3cfa/aps201223f1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e529/4010374/0dc19f16df86/aps201223f2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e529/4010374/41652ffb50a0/aps201223f3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e529/4010374/eb2bf890387e/aps201223f4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e529/4010374/cc5c79908df8/aps201223f5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e529/4010374/d996dc85ab54/aps201223f6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e529/4010374/bfc1e40979bc/aps201223f7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e529/4010374/8d359832ea2f/aps201223f8.jpg

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