基于杆状病毒-昆虫细胞表达系统的猪脑心肌炎病毒样颗粒的表达及鉴定

doi:10.11843/j.issn.0366-6964.2024.10.028

Abstract

Abstract:

Porcine encephalomyocarditis is a viral zoonotic disease caused by encephalomyocarditis virus and characterized by porcine encephalitis and myocarditis, which can infect a variety of mammals. Currently, there is no effective vaccine in China, so developing a safe and effective vaccine is an effective measure to prevent and control this disease. In this study, virus-like particals of porcine encephalomyocarditis virus were assembled by using the baculovirus-insect cell expression system with P1 and 3CD co-expression strategy, and the immune response effect of the virus in mice was preliminatively evaluated. The results showed that the specific antibody titer of mice in the immune group reached more than 1∶192, the neutralizing antibody titer reached more than 1∶64, and the combination with ISA206 adjuvant can induce higher antibody levels. The levels of IL-4, IFN-γ, IL-5 and TNF-α cytokines all reached 20 pg·mL^-1, and were significantly higher than those in PBS group. Therefore, the EMCV virus-like particle vaccine obtained in this study can induce the body to produce good cellular and humoral immune responses, and can also induce the production of inflammatory cytokines, which helps to stimulate the body's immune response, enhance the protective effect of the vaccine, and provide theoretical basis and technical reserve for the prevention and control of EMCV.

Key words: baculovirus-insect cell expression system, encephalomyocarditis virus, virus-like particles

CLC Number:

S852.65

Yunhang WANG, Wei JING, Suyu MU, Shiqi SUN, Huichen GUO, Manyuan BAI. Expression and Identification of Porcine Encephalomyocarditis Virus-like Particles Based on Baculovirus-insect Cell Expression System[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(10): 4571-4578.

Figures/Tables 5

Fig. 1

Fig. 2

Fig. 3

Fig. 4

Fig. 5

References 21

1	李丽敏, 邹云婧, 郝雪飘, 等. 脑心肌炎病毒感染神经细胞的基因表达谱分析[J]. 畜牧兽医学报, 2019, 50 (8): 1709- 1714. doi: 10.11843/j.issn.0366-6964.2019.08.021
	LI L M , ZOU Y J , HAO X P , et al. Analysis on gene expression profiling in nerve cells infected by encephalomyocarditis virus[J]. Acta Veterinaria et Zootechnica Sinica, 2019, 50 (8): 1709- 1714. doi: 10.11843/j.issn.0366-6964.2019.08.021
2	韩若婵. 猪脑心肌炎病毒2A蛋白结合annexin A2抑制细胞凋亡的分子机制[D]. 杨凌: 西北农林科技大学, 2022.
	HAN R C. Molecular mechanism of inhibition of apoptosis by encephalomyocarditis virus 2A protein binding with Annexin A2[D]. Yangling: Northwest A&F University, 2022. (in Chinese)
3	WANG Z , YU H L , ZHUANG W Y , et al. Cell pyroptosis in picornavirus and its potential for treating viral infection[J]. J Med Virol, 2022, 94 (8): 3570- 3580. doi: 10.1002/jmv.27813
4	BILLINIS C . Encephalomyocarditis virus infection in wildlife species in Greece[J]. J Wildl Dis, 2009, 45 (2): 522- 526. doi: 10.7589/0090-3558-45.2.522
5	ZHANG H X , LI S J , SU J X , et al. Construction, expression and assemble of EMCV VLPs and their potency evaluation[J]. Virology, 2023, 584, 1- 8. doi: 10.1016/j.virol.2023.04.011
6	FUENMAYOR J , GÒDIA F , CERVERA L . Production of virus-like particles for vaccines[J]. Nat Biotechnol, 2017, 39, 174- 180.
7	KIM C , KIM J D , SEO S U . Nanoparticle and virus-like particle vaccine approaches against SARS-CoV-2[J]. J Microbiol, 2022, 60 (3): 335- 346. doi: 10.1007/s12275-022-1608-z
8	CHAMBERS A C , AKSULAR M , GRAVES L P , et al. Overview of the baculovirus expression system[J]. Curr Protoc Protein Sci, 2018, 91, 5.4.1- 5.4.6.
9	WEI J , ZHANG H X , LI X R , et al. Transcriptional profiling of host cell responses to encephalomyocarditis virus (EMCV)[J]. Virol J, 2017, 14 (1): 45. doi: 10.1186/s12985-017-0718-4
10	KUSHNIR N , STREATFIELD S J , YUSIBOV V . Virus-like particles as a highly efficient vaccine platform: diversity of targets and production systems and advances in clinical development[J]. Vaccine, 2012, 31 (1): 58- 83. doi: 10.1016/j.vaccine.2012.10.083
11	JENNINGS G T , BACHMANN M F . The coming of age of virus-like particle vaccines[J]. Biol Chem, 2008, 389 (5): 521- 536. doi: 10.1515/BC.2008.064
12	CIMICA V , GALARZA J M . Adjuvant formulations for virus-like particle (VLP) based vaccines[J]. Clin Immunol, 2017, 183, 99- 108. doi: 10.1016/j.clim.2017.08.004
13	BOIGARD H , ALIMOVA A , MARTIN G R , et al. Zika virus-like particle (VLP) based vaccine[J]. PLoS Negl Trop Dis, 2017, 11 (5): e0005608. doi: 10.1371/journal.pntd.0005608
14	PEREZ E M , FOLEY J , TISON T , et al. Novel Epstein-Barr virus-like particles incorporating gH/gL-EBNA1 or gB-LMP2 induce high neutralizing antibody titers and EBV-specific T-cell responses in immunized mice[J]. Oncotarget, 2017, 8 (12): 19255- 19273. doi: 10.18632/oncotarget.13770
15	HARDING C V , SONG R . Phagocytic processing of exogenous particulate antigens by macrophages for presentation by class Ⅰ MHC molecules[J]. J Immunol, 1994, 153 (11): 4925- 4933. doi: 10.4049/jimmunol.153.11.4925
16	KOVACSOVICS-BANKOWSKI M , CLARK K , BENACERRAF B , et al. Efficient major histocompatibility complex class Ⅰ presentation of exogenous antigen upon phagocytosis by macrophages[J]. Proc Natl Acad Sci U S A, 1993, 90 (11): 4942- 4946. doi: 10.1073/pnas.90.11.4942
17	AMBUHL P M , TISSOT A C , FULURIJA A , et al. A vaccine for hypertension based on virus-like particles: preclinical efficacy and phase Ⅰ safety and immunogenicity[J]. J Hypertens, 2007, 25 (1): 63- 72. doi: 10.1097/HJH.0b013e32800ff5d6
18	BRAUN M , JANDUS C , MAURER P , et al. Virus-like particles induce robust human T-helper cell responses[J]. Eur J Immunol, 2012, 42 (2): 330- 340. doi: 10.1002/eji.201142064
19	GOLDINGER S M , DUMMER R , BAUMGAERTNER P , et al. Nano-particle vaccination combined with TLR-7 and -9 ligands triggers memory and effector CD8? T-cell responses in melanoma patients[J]. Eur J Immunol, 2012, 42 (11): 3049- 3061. doi: 10.1002/eji.201142361
20	SPEISER D E , SCHWARZ K , BAUMGAERTNER P , et al. Memory and effector CD8 T-cell responses after nanoparticle vaccination of melanoma patients[J]. J Immunother, 2010, 33 (8): 848- 858. doi: 10.1097/CJI.0b013e3181f1d614
21	ABDULRAHMAN W , RADU L , GARZONI F , et al. The production of multiprotein complexes in insect cells using the baculovirus expression system[J]. Methods Mol Biol, 2015, 1261, 91- 114.

[1]	LI Shuo, ZHANG Yun, BAI Manyuan, ZHAO Ruichong, SONG Hetao, MU Suyu, TENG Zhidong, DONG Hu, MA E'ning, SUN Shiqi, GUO Huichen, YIN Shuanghui. Immunogenicity Evaluation of Biomineralized Foot-and-mouth Disease Virus-like Particles Vaccine [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(4): 1598-1607.
[2]	ZHANG Jiaqi, JIA Xining, ZHOU Qun, SONG Xin, ZHU Chenxi, LI Gege, ZHANG Bin. Construction of BVDV-1 Virus-like Particles based on E2 Protein and Evaluation of Its Immune Efficacy in Mice [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(12): 5143-5153.
[3]	HAN Weijian, ZHANG Junjuan, ZHANG Yiming, WANG Jiaxin, LI Limin. Cytokine Responses of Bone Marrow-derived Mast Cells to FMDV-VLPs via Mannose Receptors [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(11): 3917-3926.
[4]	LIU Ruyue, FAN Jinghui, LIU Tao, YUAN Junhui, LI Qingyan, ZHAI Xianghe, ZUO Yuzhu. Construction and Identification of Hepatitis B Core Antigen Virus-like Particles Carrying PEDV S1 Epitope [J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(6): 1760-1764.
[5]	SHI Lei, ZHOU Yahong, TIAN Zhancheng, GAO Shandian, DU Junzheng, GUAN Guiquan, LIU Guangyuan, LUO Jianxun, YIN Hong. Generation of Genotype Ⅰ Japanese Encephalitis Virus Virus-like Particles and Evaluation of Its Immunogenicity in Mice [J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(6): 1391-1398.
[6]	LI Limin, ZOU Yunjing, HAO Xuepiao, ZHAO Xinghua, LI Ruiwen, SUN Jiguo, YUAN Wanzhe. Analysis on Gene Expression Profiling in Nerve Cells Infected by Encephalomyocarditis Virus [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(8): 1709-1714.
[7]	LUO Shang-xing, SHEN Xiao-qiang, GU Wen-yuan, FAN Jing-hui, DI Jing-mei, LIU Bao-jing, DAI Fei, KONG Yuan-yuan, CHANG Yan-yan, ZUO Yu-zhu. Studies on Presentation of Recombinant Parvovirus-like Particles by Porcine Splenic Dendritic Cells [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(3): 508-514.
[8]	SHEN Xiao-qiang，ZUO Yu-zhu，DI Jing-mei，GU Wen-yuan，FAN Jing-hui. The Influence of Antibodies against Key Factors of Different Endocytic Pathways on Dendritic Cells Uptake Recombinant Parvovirus-like Particles [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(9): 1924-1930.
[9]	CHANG Hong-tao，LIU Hui-min，CHEN Lu，YANG Xia，ZHAO Jun，WANG Xin-wei，YAO Hui-xia，WANG Chuan-qing. Isolation，Identification and Full Length Genome Sequence Analysis of EMCV from Different Pig Source [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(6): 967-973.
[10]	GE Sheng-qiang， ZHAO Ming，WU Xiao-dong，WANG Jing-jing，LIU Hua-lei，BAO Jing-yue，WANG Zhi-liang. Progress of Newcastle Disease Virus-like Particles [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(5): 686-691.
[11]	ZHANG Guoqing;ZHU Shu;GE Xinna;GUO Xin;CHEN Yanhong; ZHA Zhenlin;YANG Hanchun. Construction of Fulllength Infectious Clone for Encephalomyocarditis Virus BJC3 and Identification of Rescued Virus [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2009, 40(9): 1350-1357.
[12]	ZHAO Ting;ZHANG Jialong;GE Xinna;GUO Xin;ZHOU Lei;CHEN Yanhong;ZHA Zhenlin;YANG Hanchun. Comparative Analysis of the Genomes of Encephalomyocarditis Virus Isolates from Porcine and Mouse Origin [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2009, 40(6): 873-878.
[13]	PAN Qun-xing;HE Kong-wang;WANG Yong-shan;WEN Li-bin;MAO Ai-hua;GUO Rong-li. Construction and Identification of Recombinant Porcine Parvoviruslike Particles Formed by the Hybrid VP2 Protein Carrying Immunoreactive Epitope of Porcine Circovirus Type 2 [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2008, 39(9): 1245-1250.

Expression and Identification of Porcine Encephalomyocarditis Virus-like Particles Based on Baculovirus-insect Cell Expression System

RichHTML

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 5

References 21

Related Articles 13

Recommended Articles

Metrics

Comments