基因Ⅰ型乙型脑炎病毒病毒样颗粒的制备及其对小鼠的免疫原性分析

doi:10.11843/j.issn.0366-6964.2020.022

Abstract

Abstract: Decreasing the circulation of the Japanese encephalitis virus (JEV) in pigs is critical for blocking JEV infection in human. In view of the urgency of JE prevention and control in China and the uncertainty of the commercial Genotype Ⅲ JEV attenuated vaccine to the immuno-protection of partial Genotype Ⅰ JEV (GⅠ JEV) endemic strains, it is urgent to develop a safe, effective and low-cost GⅠ JEV vaccine for pigs. The expression cassette of JEV prME gene was cloned into the baculovirus shuttle vector to generate a recombinant bacmid (Bacmid-prME), the recombinant baculovirus was produced by transfecting Sf9 cells with Bacmid-prME. The accurate expression of the recombinant prME protein was confirmed by Western blot and immunofluorescence analysis, and the formation of Virus-Like Particles (VLPs) was verified through observing under Electronic microscopy. The immunogenicity of GⅠ JEV VLPs was further evaluated in mice. The recombinant prME protein was efficiently expressed in Sf9 cells and assembled into VLPs. GⅠ JEV VLPs induced the higher neutralizing antibody titer in mice. In our study, GⅠ JEV VLPs obtained in the baculovirus expression system provides a material basis for the development of a safe and effective JEV subunit vaccine.

Key words: Japanese encephalitis virus, baculovirus, virus-like particles, immunogenicity

CLC Number:

S852.659.6

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.

References 30

[1]	CAMPBELL G L, HILLS S L, FISCHER M, et al. Estimated global incidence of Japanese encephalitis:a systematic review[J]. Bull World Health Organ, 2015, 89(10):766-774E.
[2]	VAN DEN HURK A F, RITCHIE S A, JOHANSEN C A, et al. Domestic pigs and Japanese encephalitis virus infection, Australia[J]. Emerg Infect Dis, 2014, 14(11):1736-1738.
[3]	MIN J G, XUE M. Progress in studies on the overwintering of the mosquito Culex tritaeniorhynchus[J]. Southeast Asian J Trop Med Public Health, 1996, 27(4):810-817.
[4]	SOLOMON T, VAUGHN D W. Pathogenesis and clinical features of Japanese encephalitis and West Nile virus infections[J]. Curr Top Microbiol Immunol, 2002, 267:171-194.
[5]	HEGDE N R, GORE M M. Japanese encephalitis vaccines:Immunogenicity, protective efficacy, effectiveness, and impact on the burden of disease[J]. Hum Vaccin Immunother, 2017, 13(6):1320-1337.
[6]	成依依, 周红宁. 我国流行性乙型脑炎病毒基因型研究进展[J]. 中国病原生物学杂志, 2018, 13(12):1413-1419.CHENG Y Y, ZHOU H N. Advances in the study of Japanese encephalitis virus genotyping in China[J]. Journal of Pathogen Biology, 2018, 13(12):1413-1419. (in Chinese)
[7]	PARANJPE S, BANERJEE K. Phylogenetic analysis of the envelope gene of Japanese encephalitis virus[J]. Virus Res, 1996, 42(1-2):107-117.
[8]	GAO X Y, LIU H, LI M H, et al. Insights into the evolutionary history of Japanese encephalitis virus (JEV) based on whole-genome sequences comprising the five genotypes[J]. Virol J, 2015, 12:43.
[9]	GAO X Y, LIU H, WANG H Y, et al. Southernmost Asia is the source of Japanese encephalitis virus (genotype 1) diversity from which the viruses disperse and evolve throughout Asia[J]. PLoS Negl Trop Dis, 2013, 7(9):e2459.
[10]	PAN X L, LIU H, WANG H Y, et al. Emergence of genotype I of Japanese encephalitis virus as the dominant genotype in Asia[J]. J Virol, 2011, 85(19):9847-9853.
[11]	LIU X Y, YU Y X, LI M G, et al. Study on the protective efficacy of SA14-14-2 attenuated Japanese encephalitis against different JE virus isolates circulating in China[J]. Vaccine, 2011, 29(11):2127-2130.
[12]	WEI J C, WANG X, ZHANG J J, et al. Partial cross-protection between Japanese encephalitis virus genotype Ⅰ and Ⅲ in mice[J]. PLoS Negl Trop Dis, 2019, 13(8):e0007601.
[13]	CAO L, FU S H, GAO X Y, et al. Low protective efficacy of the current Japanese encephalitis vaccine against the emerging genotype 5 Japanese Encephalitis virus[J]. PLoS Negl Trop Dis, 2016, 10(5):e0004686.
[14]	DONALDSON B, LATEEF Z, WALKER G F, et al. Virus-like particle vaccines:immunology and formulation for clinical translation[J]. Expert Rev Vaccines, 2018, 17(9):833-849.
[15]	张艳芳, 杜瑞坤, 黄少梅, 等. 乙型脑炎病毒样颗粒疫苗的制备及其免疫保护效率的初步评价[J]. 病毒学报, 2016, 32(2):150-155.ZHANG Y F, DU R K, HUANG S M, et al. Generation of Japanese encephalitis virus-like particle vaccine and preliminary evaluation of its protective efficiency[J]. Chinese Journal of Virology, 2016, 32(2):150-155. (in Chinese)
[16]	HUA R H, LI Y N, CHEN Z S, et al. Generation and characterization of a new mammalian cell line continuously expressing virus-like particles of Japanese encephalitis virus for a subunit vaccine candidate[J]. BMC Biotechnol, 2014, 14:62.
[17]	于瑞明, 田占成, 独军政, 等. 基因Ⅰ型乙脑病毒EDⅢ结构域蛋白的原核表达及其抗体制备与鉴定[J]. 中国兽医科学, 2017, 47(12):1475-1480.YU R M, TIAN Z C, DU J Z, et al. Identification of EDⅢ protein from the genotype Ⅰ Japanese encephalitis virus[J]. Veterinary Science in China, 2017, 47(12):1475-1480. (in Chinese)
[18]	周亚红, 田占成, 于瑞明, 等. IFN-λ1作为乙型脑炎病毒DNA疫苗佐剂的免疫效应研究[J]. 中国兽医科学, 2019, 49(7):841-846.ZHOU Y H, TIAN Z C, YU R M, et al. Immune effect of IFN-λ1 as adjuvant for Japanese encephalitis virus DNA vaccine[J]. Chinese Veterinary Science, 2019, 49(7):841-846. (in Chinese)
[19]	HSU S T, CHANG L C, LIN S Y, et al. The effect of vaccination with a live attenuated strain of Japanese encephalitis virus on stillbirths in swine in Taiwan[J]. Bull World Health Organ, 1972, 46(4):465-471.
[20]	FAN Y C, CHIU H C, CHEN L K, et al. Formalin inactivation of Japanese encephalitis virus vaccine alters the antigenicity and immunogenicity of a neutralization epitope in envelope protein domain III[J]. PLoS Neg Trop Dis, 2015, 9(10):e0004167.
[21]	冯瑶, 岳辉喜, 刘洪明, 等. 基因Ⅰ型猪乙型脑炎灭活疫苗免疫原性评价[J]. 畜牧兽医学报, 2017, 48(5):889-895.FENG Y, YUE H X, LIU H M, et al. The immunogenicity evaluation of inactivated vaccines of swine Japanese encephalitis virus genetype I[J]. Acta Veterinaria et Zootechnica Sinica, 2017, 48(5):889-895. (in Chinese)
[22]	MANSFIELD K L, HERNÁNDEZ-TRIANA L M, BANYARD A C, et al. Japanese encephalitis virus infection, diagnosis and control in domestic animals[J]. Vet Microbiol, 2017, 201:85-92.
[23]	ZLATKOVIC J, STIASNY K, HEINZ F X. Immunodominance and functional activities of antibody responses to inactivated West Nile virus and recombinant subunit vaccines in mice[J]. J Virol, 2011, 85(5):1994-2003.
[24]	KOJIMA A, YASUDA A, ASANUMA H, et al. Stable high-producer cell clone expressing virus-like particles of the Japanese encephalitis virus e protein for a second-generation subunit vaccine[J]. J Virol, 2003, 77(16):8745-8755.
[25]	HEINZ F X, ALLISON S L. Structures and mechanisms in flavivirus fusion[J]. Adv Virus Res, 2000, 55:231-269.
[26]	KONISHI E, FUJII A, MASON P W. Generation and characterization of a mammalian cell line continuously expressing Japanese encephalitis virus subviral particles[J]. J Virol, 2001, 75(5):2204-2212.
[27]	YAMAJI H, SEGAWA M, NAKAMURA M, et al. Production of Japanese encephalitis virus-like particles using the baculovirus-insect cell system[J]. J Biosci Bioeng, 2012, 114(6):657-662.
[28]	GASTAMINZA P, DRYDEN K A, BOYD B, et al. Ultrastructural and biophysical characterization of hepatitis C virus particles produced in cell culture[J]. J Virol, 2010, 84(21):10999-11009.
[29]	KUHN R J, ZHANG W, ROSSMANN M G, et al. Structure of dengue virus:implications for flavivirus organization, maturation, and fusion[J]. Cell, 2002, 108(5):717-725.
[30]	REN X J, FU S H, DAI P F, et al. Pigsties near dwellings as a potential risk factor for the prevalence of Japanese encephalitis virus in adult in Shanxi, China[J]. Infect Dis Poverty, 2017, 6(1):100.

Generation of Genotype Ⅰ Japanese Encephalitis Virus Virus-like Particles and Evaluation of Its Immunogenicity in Mice

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