猪瘟病毒E2蛋白纳米颗粒的制备及在家兔上的免疫原性分析

doi:10.11843/j.issn.0366-6964.2025.05.028

畜牧兽医学报 ›› 2025, Vol. 56 ›› Issue (5): 2301-2311.doi: 10.11843/j.issn.0366-6964.2025.05.028

猪瘟病毒E2蛋白纳米颗粒的制备及在家兔上的免疫原性分析

张晓玲¹^,²(), 何兴林¹^,², 张梦迪¹^,³, 李鹏飞¹^,², 孙玉梅¹^,², 马海龙¹^,³, 朱红梅¹, 张梦佳¹^,³^,*(), 李文涛¹^,²^,³^,⁴^,⁵^,*()

1. 华中农业大学动物医学院，武汉 430070
2. 生猪健康养殖协同创新中心实验室，武汉 430070
3. 湖北洪山实验室，武汉 430070
4. 农业微生物资源发掘与利用全国重点实验室，武汉 430070
5. 华中农业大学动物疫病诊断中心，武汉 430070

收稿日期:2024-07-09 出版日期:2025-05-23 发布日期:2025-05-27
通讯作者: 张梦佳,李文涛 E-mail:zxl15136139755@163.com;zhangmengjia0210@mail.hzau.edu.cn;wentao@mail.hzau.edu.cn
作者简介:张晓玲(2000-)，女，河南洛阳人，硕士，主要从事亚单位疫苗研究，E-mail：zxl15136139755@163.com
基金资助:
国家现代农业产业技术体系(CARS-35)；影子科技-华中农大健康食品产业研究院项目-猪场重大疫病监测与预警创新平台建设(IRIFH202209)

Preparation of E2 Protein Nanoparticles from Classical Swine Fever Virus and the Immunogenicity Study in Rabbit

ZHANG Xiaoling¹^,²(), HE Xinglin¹^,², ZHANG Mengdi¹^,³, LI Pengfei¹^,², SUN Yumei¹^,², MA Hailong¹^,³, ZHU Hongmei¹, ZHANG Mengjia¹^,³^,*(), LI Wentao¹^,²^,³^,⁴^,⁵^,*()

1. College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China
2. Collaborative Innovation Center Laboratory for Pig Health Breeding, Wuhan 430070, China
3. Hubei Hongshan Laboratory, Wuhan 430070, China
4. National Key Laboratory of Agricultural Microbiology, Wuhan 430070, China
5. Animal Disease Diagnostic Center, Huazhong Agricultural University, Wuhan 430070, China

Received:2024-07-09 Online:2025-05-23 Published:2025-05-27
Contact: ZHANG Mengjia, LI Wentao E-mail:zxl15136139755@163.com;zhangmengjia0210@mail.hzau.edu.cn;wentao@mail.hzau.edu.cn

摘要/Abstract

摘要：

本研究旨在开发一种猪瘟E2纳米颗粒疫苗，并对其免疫原性进行分析。以猪瘟病毒石门株E2基因序列为模板，对其胞外域进行人源密码子优化，通过293F哺乳动物细胞真核表达系统制备E2、E2-pFc(E2与猪源抗体的Fc片段融合表达)、SpyTag-E2(E2与SpyTag融合表达)以及纳米骨架蛋白LS(lumazine synthase-protein A)，原核表达系统纯化纳米骨架蛋白mi3(mi3-SpyCatcher)，进一步组装成纳米颗粒LS-E2-pFc NP和mi3-E2 NP，并通过透射电镜和动态光散射试验进行鉴定，各组蛋白分别与ISA201佐剂乳化，制备亚单位疫苗。选用1.5~2.0 kg重的日本长耳大白兔35只，随机分为7组，包括空白对照组(n=3)、PBS阴性对照组(n=3)、E2组(n=5)、E2-pFc组(n=5)、LS-E2-pFc NP组(n=5)、mi3-E2 NP组(n=5)和CSFV E2商业化亚单位疫苗组(n=3)。于0、21 d进行肌肉注射免疫，42 d将CSFV C株经耳缘静脉按照100 RID₅₀ ·只^-1进行攻毒，通过阻断ELISA、中和试验、体温监测和RT-qPCR等试验评估兔的特异性抗体水平、中和抗体水平、兔体定型热反应以及脾脏病毒载量。结果显示，各组分蛋白成功表达，纯化后的蛋白条带单一，纯度较高。透射电镜可观察到粒径在10~70 nm的纳米颗粒结构，动态光散射试验观察到mi3-E2平均粒径(68 nm)大于mi3(42.73 nm)，LS-E2-pFc平均粒径(42.4 nm)大于LS(13 nm)。免疫试验结果表明，E2、E2-pFc、LS-E2-pFc NP、mi3-E2 NP疫苗和商业化亚单位疫苗均有良好的免疫效果，攻毒前7 d，血清中特异性抗体阻断率均在89%以上，组间差异不显著(P>0.05)。攻毒后，PBS阴性对照组出现典型的兔体定型热反应，其余免疫组的体温变化不明显；攻毒后7 d，E2、E2-pFc、LS-E2-pFc NP、mi3-E2 NP疫苗和商业化亚单位疫苗组的中和抗体效价分别为1 ∶10 392.4、1 ∶37 168.4、1 ∶75 473.6、1 ∶20 201.8、1 ∶2 407，其中，LS-E2-pFc NP组的中和抗体水平最高。RT-qPCR结果显示，E2组有一只兔检出低拷贝的病毒载量，拷贝数为5.83 copies ·μL^-1，其余免疫组未检到病毒载量。以上结果显示LS-E2-pFc NP免疫组效果最好，中和抗体水平最高。这种“标记”疫苗可以对自然感染和免疫接种的动物进行鉴别诊断，有助于猪场净化猪瘟。

关键词: 猪瘟病毒, E2亚单位疫苗, 自组装纳米疫苗, 中和抗体

Abstract:

The aim of this study was to develop a nanoparticle vaccine targeting the E2 protein of classical swine fever virus (CSFV) and analyze its immunogenicity. Using the E2 gene sequence of Shimen strain of CSFV as template, the human codon was optimized in its extracellular domain. E2, E2-pFc (fusion expression of E2 with Fc fragment of porcine antibody), SpyTag-E2 (fusion expression of E2 with SpyTag) and the nanoskeleton protein LS (Lumazine Synthase-protein A) were prepared by 293F mammal cell eukaryotic expression system. Nanoskeleton protein mi3 (mi3-SpyCatcher) was purified by prokaryotic expression system and then LS and mi3 assembled into nanoparticles LS-E2-pFc NP and mi3-E2 NP, which were identified by transmission electron microscopy and dynamic light scattering test, and each protein was emulsified with ISA201 adjuvant to prepare subunit vaccines. Thirty-five Japanese white rabbits (1.5-2.0 kg) were randomly divided into 7 groups. Blank control group (n=3), PBS negative control group (n=3), E2 group (n=5), E2-pFc group (n=5), LS-E2-pFc NP group (n=5), mi3-E2 NP group (n=5), and CSFV E2 commercial subunit vaccine group (n=3). On 0 d and 21 d, intramuscular immunization was administered, and on 42 d, CSFV C strain was challenged with 100 RID₅₀ via ear vein. The specific antibody level, neutralizing antibody level, body temperature response and spleen viral load of rabbits were evaluated by blocking ELISA, neutralizing test, temperature monitoring and RT-qPCR. The results showed the successful expression of all component proteins, with single-band profiles and high purity. The average particle size of mi3-E2 nanoparticles (68 nm) was larger than mi3 (42.73 nm), and LS-E2-pFc nanoparticles (42.4 nm) was larger than LS (13 nm). Immunization results showed that E2, E2-pFc, LS-E2-pFc NP, mi3-E2 NP vaccine and commercial subunit vaccine all had good immune effect, and 7 days before challenge, the specific antibody blocking rate in serum was above 89%, with no significant difference between immunized groups (P>0.05). After challenge, the PBS negative control group showed typical stereotyped heat reaction, while the other immune groups showed no obvious changes in body temperature. Seven days post-challenge, the titers of neutralizing antibodies of E2, E2-pFc, LS-E2-pFc NP, mi3-E2 NP vaccine and commercial subunit vaccine groups were 1:10 392.4, 1:37 168.4, 1:75 473.6, 1:20 201.8 and 1:2 407, respectively, with the LS-E2-pFc NP group showing the highest neutralizing antibody levels. RT-qPCR results showed that one rabbit in the E2 group had a low viral load of 5.83 copies ·μL^-1, while no viral load was detected in other immunized groups. The above results showed that LS-E2-pFc NP immunized group had the best effect and the highest titer of neutralizing antibody. This "marker" vaccine allows for differential diagnosis of naturally infected and vaccinated animals, helping to eradicate CSF in swine farms.

Key words: classical swine fever virus, E2 subunit vaccine, self-assembled nano vaccines, neutralizing antibody

中图分类号:

S852.651

张晓玲, 何兴林, 张梦迪, 李鹏飞, 孙玉梅, 马海龙, 朱红梅, 张梦佳, 李文涛. 猪瘟病毒E2蛋白纳米颗粒的制备及在家兔上的免疫原性分析[J]. 畜牧兽医学报, 2025, 56(5): 2301-2311.

ZHANG Xiaoling, HE Xinglin, ZHANG Mengdi, LI Pengfei, SUN Yumei, MA Hailong, ZHU Hongmei, ZHANG Mengjia, LI Wentao. Preparation of E2 Protein Nanoparticles from Classical Swine Fever Virus and the Immunogenicity Study in Rabbit[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(5): 2301-2311.

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参考文献 32

1	GREISER-WILKEI,MOENNIGV.Vaccination against classical swine fever virus: limitations and new strategies[J].Anim Health Res Rev,2004,5,223-226. doi: 10.1079/AHR200472
2	KADENV,LANGEE,STEYERH,et al.Role of birds in transmission of classical swine fever virus[J].J Vet Med B Infect Dis Vet Public Health,2003,50,357-359. doi: 10.1046/j.1439-0450.2003.00670.x
3	GRAHAMS P,EVERETTH E,HAINESF J,et al.Challenge of pigs with classical swine fever viruses after C-strain vaccination reveals remarkably rapid protection and insights into early immunity[J].PloS one,2012,7,e29310. doi: 10.1371/journal.pone.0029310
4	FERRARIM.A tissue culture vaccine with lapinized chinese (LC) strain of hog cholera virus (HCV)[J].Comp Immunol Microbiol Infect Dis,1992,15(3):221-228. doi: 10.1016/0147-9571(92)90095-9
5	HANY,XIEL,YUANM,et al.Development of a marker vaccine candidate against classical swine fever based on the live attenuated vaccine C-strain[J].Vet Microbiol,2020,247,108741. doi: 10.1016/j.vetmic.2020.108741
6	SUNS,CAIY,SONGT Z,et al.Interferon-armed RBD dimer enhances the immunogenicity of RBD for sterilizing immunity against SARS-CoV-2[J].Cell Res,2021,31,1011-1023. doi: 10.1038/s41422-021-00531-8
7	HSIAY,BALEJ B,GONENS,et al.Corrigendum: Design of a hyperstable 60-subunit protein icosahedron[J].Nature,2016,540(7631):150.
8	SONGH,ABDULLAHS W,PEIC,et al.Self-assembling E2-based nanoparticles improve vaccine thermostability and protective immunity against CSFV[J].Int J Mol Sci,2024,25,596. doi: 10.3390/ijms25010596
9	XUH,WANGY,HANG,et al.Identification of E2 with improved secretion and immunogenicity against CSFV in piglets[J].BMC Microbiol,2020,20,26. doi: 10.1186/s12866-020-1713-2
10	KEEBLEA H,BANERJEEA,FERLAM P,et al.Evolving accelerated amidation by SpyTag/SpyCatcher to analyze membrane dynamics[J].Angew Chem Int Ed Engl,2017,56,16521-16525. doi: 10.1002/anie.201707623
11	BRUUNT U J,ANDERSSONA C,DRAPERS J,et al.Engineering a rugged nanoscaffold to enhance plug-and-display vaccination[J].ACS Nano,2018,12,8855-8866. doi: 10.1021/acsnano.8b02805
12	LIUZ H,XUH L,HANG W,et al.A self-assembling nanoparticle: Implications for the development of thermostable vaccine candidates[J].Int J Biol Macromol,2021,183,2162-2173. doi: 10.1016/j.ijbiomac.2021.06.024
13	SASAKIE,BÖHRINGERD,VAN DE WATERBEEMDM,et al.Structure and assembly of scalable porous protein cages[J].Nat Commun,2017,8,14663. doi: 10.1038/ncomms14663
14	WÖRSDÖRFERB,WOYCECHOWSKYK J,HILVERTD.Directed evolution of a protein container[J].Science,2011,331,589-592. doi: 10.1126/science.1199081
15	LIW,HULSWITR J G,WIDJAJAI,et al.Identification of sialic acid-binding function for the middle east respiratory syndrome coronavirus spike glycoprotein[J].Proc Natl Acad Sci U S A,2017,114,8508-8517.
16	王鹤儒,吴业红,何鹏,等.糖基化对疫苗免疫原性影响的研究进展[J].中国新药杂志,2024,33(3):241-246.
	WANGH R,WUY H,HEP,et al.Research progress on the effect of glycosylation on vaccine immunogenicity[J].Chinese Journal of New Drugs,2024,33(3):241-246.
17	GAVRILOVB K,ROGERSK,FERNANDEZ-SAINZI J,et al.Effects of glycosylation on antigenicity and immunogenicity of classical swine fever virus envelope proteins[J].Virology,2011,420(2):135-145. doi: 10.1016/j.virol.2011.08.025
18	魏蔷,刘运超,白怡霖,等.信号肽对猪瘟病毒E2蛋白在杆状病毒表达系统中分泌表达的影响[J].畜牧兽医学报,2020,51(1):120-127. doi: 10.11843/j.issn.0366-6964.2020.01.014
	WElQ,LlUY C,BAlY L,et al.Effect of signal peptide on the secretory expression of CSFV E2 protein in baculovirus expression system[J].Acta Veterinaria et Zootechnica Sinica,2020,51(1):120-127. doi: 10.11843/j.issn.0366-6964.2020.01.014
19	朱蓓蓓. 猪瘟病毒单克隆抗体的制备及其初步应用[D]. 武汉: 华中农业大学, 2009.
	ZHU B B. Development and application of monoclonal antibodies against classical swine fever virus[D]. Wuhan: Huazhong Agricultural University, 2009. (in Chinese)
20	马帅,郑世磊,赵明,等.我国猪瘟的流行病学及疫苗研究进展[J].中国动物传染病学报,2022,30(6):211-218.
	MAS,ZHENGS L,ZHAOM,et al.Epidemiology and vaccine research progress of classical swine fever in China[J].Chinese Journal of Animal infectious Diseases,2022,30(6):211-218.
21	ZHANGH,LENGC,FENGL,et al.A new subgenotype 2.1d isolates of classical swine fever virus in China, 2014[J].Infect Genet Evo,2015,34,94-105. doi: 10.1016/j.meegid.2015.05.031
22	GONGW,LIJ,WANGZ,et al.Commercial E2 subunit vaccine provides full protection to pigs against lethal challenge with 4 strains of classical swine fever virus genotype 2[J].Vet Microbiol,2019,237,108403. doi: 10.1016/j.vetmic.2019.108403
23	LINGJ,LIUTY,TSENGYY,et al.Yeast-expressed classical swine fever virus glycoprotein E2 induces a protective immune response[J].Vet Microbiol,2009,139,369-374. doi: 10.1016/j.vetmic.2009.06.027
24	WANGW,ZHOUX,BIANY,et al.Dual-targeting nanoparticle vaccine elicits a therapeutic antibody response against chronic hepatitis B[J].Nat Nanotechnol,2020,15(5):406-416. doi: 10.1038/s41565-020-0648-y
25	BROUWERP J M,BRINKKEMPERM,MAISONNASSEP,et al.Two-component spike nanoparticle vaccine protects macaques from SARS-CoV-2 infection[J].Cell,2021,84(5):1188-1200.
26	COHENA A,GNANAPRAGASAMP N P,LEEY E,et al.Mosaic nanoparticles elicit cross-reactive immune responses to zoonotic coronaviruses in mice[J].Science,2021,371(6530):735-741.
27	BOYOGLU-BARNUMS,ELLISD,GILLESPIERA,et al.Quadrivalent influenza nanoparticle vaccines induce broad protection[J].Nature,2021,592(7855):623-628.
28	ZHANGX,WUS,LIUJ,et al.A mosaic nanoparticle vaccine elicits potent mucosal immune response with significant cross-protection activity against multiple SARS-CoV-2 sublineages[J].Adv Sci (Weinh),2023,10(27):e2301034.
29	LIUX,ZHAOT,WANGL,et al.A mosaic influenza virus-like particles vaccine provides broad humoral and cellular immune responses against influenza A viruses[J].NPJ Vaccines,2023,8(1):132.
30	GANGESL,NÚÑEZJI,SOBRINOF,et al.Recent advances in the development of recombinant vaccines against classical swine fever virus: cellular responses also play a role in protection[J].Vet J,2008,177,169-177.
31	韩玉莹,谢利豹,李永锋,等.表达增强型绿色荧光蛋白的报告猪瘟病毒C株的构建及在家兔中生物学特性评价[J].畜牧兽医学报,2020,51(7):1699-1709. doi: 10.11843/j.issn.0366-6964.2020.07.022
	HANY Y,XIEL B,LIY F,et al.Generation and characterization in rabbits of a reporter classical swine fever virus vaccine C-strain expressing the enhanced green fluorescent protein[J].Acta Veterinaria et Zootechnica Sinica,2020,51(7):1699-1709. doi: 10.11843/j.issn.0366-6964.2020.07.022
32	CAOZ,ZHANGH,YANGQ,et al.Establishment of a method for evaluation of the efficacy of a classical swine fever virus subunit vaccine in rabbits[J].Am J Vet Res,2020,81,521-526.

引物名称 Primer name	引物序列(5′→3′) Primer sequence
E2-F	F: CGCTTCCGTGCTAGCAGACCGCCTCGCATGCAAGGAAGACTA
E2-R	R: GGCTCCAACTCCCTCCTCCGTAGTCGCTGTGGCGGTCGGTTG
pCAGGS-F	F: AGGAGGGAGTTGGAGCCA
pCAGGS-R	R: GTCTGCTAGCACGGAAGC
SpyTag-E2-F	F: CGCTTCCGTGCTAGCAGACGCTCACATCGTGATGGTGGACG
SpyTag-E2-R	R: GGCTCCAACTCCCTCCTCCGTAGTCGCTGTGGCGGTCGGTTG
pFRT-F	F: GACGGCCGCGAGCGGCCGCCTCGAGTCTAGAGGGCCCG
pFRT-R	R: GTCTGCTAGCACGGAAGCGACC
E2-pFc-E2-F-1	F: TCGCTTCCGTGCTAGCAGACCGCCTCGCATGCAAGGAAGAC
E2-pFc-E2-R-1	R: CTGGGCACCAGCTTAATTAAGTAGTCGCTGTGGCGGTCGGT
E2-pFc-E2-R-2	R: GCCGCCGGAGCCTCTGGGCACCAGCTTAA
pFc-F-1	F: GCCCAGAGGCTCCGGCGGCGGAACTAAAACTAAACCCCCTT
pFc-R-1	R: CGGCCGCTCGCGGCCGTCATTTTCCCTGAGTTTTGGAGATG
DL-F	F: TACAGGACAGTCGTCAGTAGTTCGA
DL-R	R: CCGCTAGGGTTAAGGTGTGTCT
probe	FAM-CCCACCTCGAGATGCTATGTGGACGA-TAMRA

蛋白名称 Protein name	蛋白的氨基酸数量/aa Amino acids number of protein	糖基化位点/个 Glycosylation sites number	糖基化修饰前大小/ku Molecular mass before glycosylation modification	糖基化修饰后大小/ku Molecular mass after glycosylation modification	产量/(mg·L^-1) Production
LS-protein A	243	0	26.73	26.73	5.580
SpyTag-E2	398	7	43.78	61.28	8.710
E2	370	7	40.70	58.20	14.860
E2-pFc	579	8	63.69	83.69	31.52

组别 Groups	兔数量 No. of rabbits	C株拷贝数/(copies·μL^-1) Viral RNA load of C-strain
空白对照组	A1	-
Blank control group	A2	-
	A3	-
PBS阴性对照组	B1	58.94
Negative control group (PBS)	B2	49.58
	B3	85.39
E2组	C1	5.83
E2 group	C2	-
	C3	-
	C4	-
	C5	-
E2-pFc组	D1	-
E2-pFc group	D2	-
	D3	-
	D4	-
	D5	-
mi3-E2组	E1	-
mi3-E2 group	E2	-
	E3	-
	E4	-
	E5	-
LS-E2-pFc组	F1	-
LS-E2-pFc group	F2	-
	F3	-
	F4	-
	F5	-
商业化亚单位疫苗组	G1	-
Commercial subunit vaccine group	G2	-
	G3	-

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猪瘟病毒E2蛋白纳米颗粒的制备及在家兔上的免疫原性分析

Preparation of E2 Protein Nanoparticles from Classical Swine Fever Virus and the Immunogenicity Study in Rabbit

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