A 型塞内卡病毒 VP2 蛋白多表位抗原免疫原性分析

doi:10.3969/j.issn.1674-7968.2023.06.016

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Abstract Senecavirus A (SVA), a new RNA virus belonging to the Picornaviridae family, which seriously endangers the development of China's pig industry. At present, there is no effective SVA vaccine. In order to study the new SVA subunit vaccine, this study was based on the viral structural protein 2 (VP2) gene sequences of SVA, and the B cell epitope sequences predicted by bioinformatic methods and the T cell epitope genes in the reported Foot-and-mouth disease virus (FMDV) 3A protein. The recombinant expression plasmids pET28a-rSVP2-B and pET28a-rSVP2-BT were constructed respectively after optimization codon and connecting in series with the flexible linkers. The recombinant tandem epitope proteins rSVP2-B and rSVP2- BT, which were expressed in prokaryotic system and purified by Ni+ ion chromatographic method, were identified by SDS-PAGE and Western blot, and then immunized BALB/c mice (Mus musculus) to evaluate their immunogenicity. The results showed that the target proteins were expressed correctly after the recombined plasmid pET28a-rSVP2-B and pET28a-rSVP2-BT were transformed into Escherichia coli BL21 (DE3) and induced by 1 mmol/L isopropy- β -D-thiogalactoside (IPTG). SDS-PAGE analysis showed that the relative molecular weights were 34 and 38 kD, respectively. Western blot results showed that rSVP2-B and rSVP2-BT had good reactivity with SVA positive sera. There was no significant difference in humoral immune levels between rSVP2-B and rSVP2-BT immunized mice with purification protein. Both of them induced strong specific IgG and IgG1 antibodies, and mainly induced humoral immune response biased towards Th2 type. The results of virus neutralization test showed that the neutralizing antibody titers induced by the two recombinant proteins ranged from 1∶64 to 1∶128. Meanwhile, mice immunized rSVP2-B and rSVP2-BT also produced strong cellular immunity. The mice spleen lymphocytes proliferation levels and interleukin-2 (IL-2) and interferon- γ (IFN- γ) expression levels in the mice vaccinated with the rSVP2-BT protein group was significantly higher than those in the rSVP2-B group. In conclusion, the recombinant tandem multi-epitope proteins rSVP2-B and rSVP2-BT with good immunogenicity were successfully prepared, and the immunogenicity of rSVP2-BT was superior to that of rSVP2-B. This study provides a theoretical basis for the development of SVA multi-epitope vaccine.

Key words： Senecavirus A (SVA) Viral structural protein 2 (VP2) Recombinant tandem epitope protein Soluble expression Immunogenicity

Received: 03 January 2023

Corresponding Authors: *weiym@gsau.edu.cn

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	Articles by authors
	TAO Shi-Yu
	RU Yi
	CHEN Jiao
	HAO Rong-Zeng
	LI Ya-Jun
	LU Bing-Zhou
	SHI Zheng-Wang
	LIU Xue-Rong
	ZHENG Hai-Xue
	WEI Yan-Ming

Cite this article:

TAO Shi-Yu,RU Yi,CHEN Jiao, et al. Immunogenicity Analysis of Multi-epitopes Antigen of Senecavirus A VP2 Protein[J]. 农业生物技术学报, 2023, 31(6): 1284-1295.

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http://journal05.magtech.org.cn/Jwk_ny/EN/10.3969/j.issn.1674-7968.2023.06.016 OR http://journal05.magtech.org.cn/Jwk_ny/EN/Y2023/V31/I6/1284

[1] 郭丽莹, 冯霞, 潘颂佳, 等. 2022. 塞内卡病毒 A 结构蛋白 T 细胞抗原表位的筛选与鉴定[J]. 中国兽医科学, 52(01): 77-84.
(Guo L Y, Feng X, Pan S J, et al. The screening and identification of T cell epitopes in the structural protein of Senecavirus A[J]. Chinese Veterinary Science, 52(01): 77-84. )
[2] 吕强. 2020. 塞内卡谷病毒中和性单克隆抗体制备及中和表位鉴定[D]. 硕士学位论文, 华中农业大学, 导师: 李祥敏 , pp. 61-62.
(Lv Q. 2020. Generation of neutralizing monoclonal antibodies against Seneca valley virus and identification of their neutralizing epitopes[D]. Thesis for M. S. , Huazhong Agricultural University, Supervisor: Li X M, pp. 61-62. )
[3] 伍春平. 2021. A 型塞内卡病毒结构蛋白 B 细胞抗原表位的鉴定[D]. 硕士学位论文, 中国农业科学院, 导师: 郑海学, pp. 42. (Wu C P. 2021. Identification of B-cell epitopes on the structural proteins of Senecavirus A[D]. The sis for M. S. , Chinese Academy of Agricultural Sciences, Supervisor: Zheng H X, pp. 42. )
[4] 姚飞. 2020. 塞内卡病毒 A VP1、VP2 蛋白 B 细胞表位的筛选与鉴定[D]. 硕士学位论文, 中国农业科学院, 导师: 潘丽, pp. 39. (Yao F. 2020. Screening and identification of B cell epitopes of VP1 and VP2 proteins of Senecavirus A [D]. Thesis for M. S. , Chinese Academy of Agricultural Sciences, Supervisor: Pan L, pp. 39. )
[5] 张涛, 王会宝, 孙燕燕, 等. 2022. A 型塞内卡病毒 VP1 蛋白单克隆抗体制备及胶体金试纸条的试制[J]. 农业生物技术学报, 30(11): 2255-2266.
(Zhang T, Wang H B, Sun Y Y, et al. 2022. Preparation of monoclonal antibody based on VP1 protein against Senecavirus A (SVA) and trial production of colloidal gold test strips for SVA detection[J]. Journal of Agricultural Biotechnology, 30(11): 2255-2266. )
[6] Blanco E, Cubillos C, Moreno N, et al. 2013. B epitope multiplicity and B/T epitope orientation influence immunogenicity of foot-and-mouth disease peptide vaccines[J]. Clinical & Developmental Immunology, 2013: 475960.
[7] Blanco E, Garcia-Briones M, Sanz-Parra A, et al. 2001. Identification of T-cell epitopes in nonstructural proteins of Foot-and-mouth disease virus[J]. Journal of Virology, 75(7): 3164-3174.
[8] Bohórquez J A, Defaus S, Muñoz-González S, et al. 2017. A bivalent dendrimeric peptide bearing a T-cell epitope from Foot-and-mouth disease virus protein 3A improves humoral response against classical Swine fever virus[J]. Virus Research, 238: 8-12.
[9] Cao L, Zhang R, Liu T T, et al. 2018. Seneca Valley virus attachment and uncoating mediated by its receptor anthrax toxin receptor 1[J]. Proceedings of the National Acade my of Sciences of the USA, 115(51): 13087-13092.
[10] Carr B V, Lefevre E A, Windsor M A, et al. 2013. CD4+ T-cell responses to Foot-and-mouth disease virus in vaccinated cattle[J]. Journal General Virology, 94 (Pt1): 97-107. Combadière B, Beaujean M, Chaudesaigues C, et al. 2019. Peptide-based vaccination for antibody responses against HIV[J]. Vaccines, 7(3): 105-119.
[11] Dvorak C M, Akkutay-Yoldar Z, Stone S R, et al. 2017. An indirect enzyme-linked immunosorbent assay for the identification of antibodies to Senecavirus A in swine[J]. BMC Veterinary Research, 13(1): 50.
[12] Fan H, Zhu H X, Li S H, et al. 2020. Identification of linear B cell epitopes on VP1 and VP2 proteins of Senecavirus A (SVA) using monoclonal antibodies[J]. Veterinary Microbiology, 247: 108753-108761.
[13] Gimenez-Lirola L G, Rademacher C, Linhares D, et al. 2016. Serological and molecular detection of Senecavirus A associated with an outbreak of swine idiopathic vesicular disease and neonatal mortality[J]. Journal of Clinical Microbiology, 54(8): 2082-2089.
[14] Hajighahramani N, Nezafat N, Eslami M, et al. 2017. Immu‐noinformatics analysis and in silico designing of a novel multi-epitope peptide vaccine against Staphylococcus au- reus[J]. Infection, Genetics and Evolution, 48: 83-94.
[15] Hales L M, Knowles N J, Reddy P S, et al. 2008. Complete genome sequence analysis of Seneca valley virus-001, a novel oncolytic picornavirus[J]. The Journal of General Virology, 89(Pt 5): 1265-1275.
[16] Leme R A, Zotti E, Alcântara B K, et al. 2015. Senecavirus A: An emerging vesicular infection in Brazilian pig herds[J]. Transboundary and Emerging Diseases, 62(6): 603-611.
[17] Liu F X, Wang Q Q, Huang Y L, et al. 2020. A 5-year review of Senecavirus A in China since its emergence in 2015[J]. Frontiers in Veterinary Science, 7: 567792.
[18] Luo D K, Wang H W, Wang Q, et al. 2022. Senecavirus A as an Oncolytic virus: Prospects, challenges and development directions[J]. Frontiers in Oncology, 12: 839536.
[19] Maggioli M F, Lawson S, de Lima M, et al. 2018. Adaptive immune responses following Senecavirus A infection in pigs[J]. Journal Virology, 92(3): e01717-17.
[20] Pasma T, Davidson S, Shaw S L. 2008. Idiopathic vesicular disease in swine in Manitoba[J]. The Canadian Veterinary Journal, 49(1): 84-85.
[21] Reddy Chichili V P, Kumar V, Sivaraman J, et al. 2012. Linkers in the structural biology of protein-protein interactions[J]. The Protein Society, 22(2): 153-67.
[22] Reddy P S, Burroughs K D, Hales L M, et al. 2007. Seneca valley virus, a systemically deliverable oncolytic picornavirus, and the treatment of neuroendocrine cancers[J]. Journal of the National Cancer Institute, 99(21): 1623-1633.
[23] Segalés J, Barcellos D, Alfieri A, et al. 2017. Senecavirus A: An emerging pathogen causing vesicular disease and mortality in pigs ?[J]. Veterinary Pathology, 54(1): 11-21.
[24] Venkataraman S, Reddy S P, Loo J, et al. 2008. Crystallization and preliminary X-ray diffraction studies of Seneca val- ley virus-001, a new member of the Picornaviridae family[J]. Acta Crystallographica Section F: Structural Biology and Crystallization Communications, 64(pt 4): 293-296.
[25] Wen W, Chen X H, Lv Q, et al. 2022. Identifcation of a conserved neutralizing epitope in Seneca valley virus VP2 protein: New insight for epitope vaccine designment[J]. Virology Journal, 19(1): 65.
[26] Zhang X L, Zhu Z X, Yang F, et al. 2018. Review of Seneca valley virus: A call for increased surveillance and research[J]. Frontiers in Microbiology, 9: 940.