不同佐剂对猪圆环病毒衣壳蛋白免疫原性的影响

doi:10.16431/j.cnki.1671-7236.2016.11.034

Abstract

Abstract:

The study was aimed to prepare vaccines with different adjuvants,and research its effects on immunogenicity.The PCV2 Cap gene with its signal peptide removed was connected to pET-28a vector,and then was induced to express,using sodium deoxycholate（DOC）and low concentration of urea to dissolve the inclusion body.Different adjuvants,such as alum-based adjuvants,liposome adjuvants,propolis adjuvants,white oil adjuvants and Freund adjuvant were prepared,together with protein purified to make up subunit vaccines,and commercial inactivated vaccine as positive control,immuning mice,and ELISA method were used to detect changes in concentrations of animal serum antibody and cytokines,evaluated the immune protective effect.Results showed that the expression product in the form of inclusion body,using the DOC could omit dissolving inclusion body protein renaturation steps,and the purification method was simple,the capsid protein obtained had high purity.ELISA assays showed that PCV2-Cap had good immunogenicity.And we found that the water system adjuvants had high immune activity,this could provide important previous experimental data for the commercialization of the subunit vaccine.

Key words: porcine circovirus; inclusion body; adjuvant; subunit vaccine

CLC Number:

WANG Li-bo, WU Run-sheng, YAN Chao, LV Tun. Effects of Different Adjuvants on the Immunogenicity of PCV2-Cap Protein[J]. , 2016, 43(11): 3037-3046.

References

[1] Fauquet C M,Mayo M A,Maniloff J,et al.Virus taxonomy:Ⅷth Report of the International Committee on Taxonomy of Viruses[M].New York:Academic Press,2005.
[2] Tischer I,Bode L,Peters D,et al.Distribution of antibodies to porcine circovirus in swine populations of different breeding farms[J].Archives of Virology,1995,140(4):737-743.
[3] Ellis J,Hassard L,Clark E,et al.Isolation of circovirus from lesions of pigs with postweaning multisystemic wasting syndrome[J].The Canadian Veterinary Journal,1998,39(1):44-51.
[4] Guo L J,Lu Y H,Wei Y W,et al.Porcine circovirus type 2(PCV2):Genetic variation and newly emerging genotypes in China[J].Virol J,2010,7(1):273-284.
[5] Finsterbusch T,Mankertz A.Porcine circoviruses——Small but powerful[J].Virus Research,2009,143(2):177-183.
[6] Hamel A L,Lin L L,Nayar G P.Nucleotide sequence of porcine circovirus associated with postweaning multisystemic wasting syndrome in pigs[J].Journal of Virology,1998,72(6):5262-5267.
[7] Mankertz A,Persson F,Mankertz J,et al.Mapping and characterization of the origin of DNA replication of porcine circovirus[J].Journal of Virology,1997,71(3):2562-2566.
[8] Mahé D,Blanchard P,Truong C,et al.Differential recognition of ORF2 protein from type 1 and type 2 porcine circoviruses and identification of immunorelevant epitopes[J].Journal of General Virology,2000,81(7):1815-1824.
[9] Fan H,Ju C,Tong T,et al.Immunogenicity of empty capsids of porcine circovius type 2 produced in insect cells[J].Veterinary Research Communications,2007,31(4):487-496.
[10] Ramon G.Sur l'augmentation anormale de l'antitoxine chez les chevaux producteurs de serum antidiphterique[J].Bull Soc Centr Med Vet,1925,101:227-234.
[11] Freund J,Casals J,Hosmer E P.Sensitization and antibody formation after injection of tubercle bacilli and paraffin oil[J].Experimental Biology and Medicine,1937,37(3):509-513.
[12] Stuart-Harris C.Adjuvant influenza vaccines[J].Bulletin of the World Health Organization,1969,41(3-5):615.
[13] Ribi E.Beneficial modification of the endotoxin molecule[J].Journal of Immunotherapy,1984,3(1):1-9.
[14] Weiner G J,Liu H M,Wooldridge J E,et al.Immunostimulatory oligodeoxynucleotides containing the CpG motif are effective as immune adjuvants in tumor antigen immunization[J].Proceedings of the National Academy of Sciences,1997,94(20):10833-10837.
[15] Vogel F.Adjuvants in perspective[J].Developments in Biological Standardization,1997,92:241-248.
[16] 肖勇翔,陈志祥,陆伟根.阳离子脂质体疫苗佐剂研究进展[J].世界临床药物,2013,34(1):41-48.
[17] 黄文,白杨,王继德,等.幽门螺杆菌热休克蛋白60脂质体疫苗的制备及其免疫预防作用[J].第一军医大学学报,2005,25(5):531-534.
[18] Nawagitgul P,Morozov I,Bolin S R,et al.Open reading frame 2 of porcine circovirus type 2 encodes a major capsid protein[J].Journal of General Virology,2000,81(9):2281-2287.
[19] 乔翠,高凤山,许崇波.猪圆环病毒2型分子致病机理及相关疾病研究进展[J].中国畜牧兽医,2012,39(4):170-173.
[20] 李海花,覃尧,张全红,等.猪圆环病毒2型Cap全基因的克隆与原核表达[J].中国畜牧兽医,2015,42(2):337-341.
[21] Chi J N等著,谢冰译.猪圆环病毒2型病毒样颗粒的制备及其在疫苗研制中的应用[J].中国畜牧兽医,2014,41(6):22.
[22] 李成文,陈咏梅,孙万邦.脂质体佐剂疫苗研究进展[J].国外医学:预防、诊断、治疗用生物制品分册,2004,26(6):255-258.
[23] Fort M,Sibila M,Pérez-Martín E,et al.One dose of a porcine circovirus 2(PCV2）sub-unit vaccine administered to 3-week-old conventional piglets elicits cell-mediated immunity and significantly reduces PCV2 viremia in an experimental model[J].Vaccine,2009,27(30):4031-4037.
[24] 王宪文.猪圆环病毒2型疫苗研究进展[J].中国畜牧兽医,2014,41(4):252-255.

Effects of Different Adjuvants on the Immunogenicity of PCV2-Cap Protein

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	HAN Bing, TANG Siqi, ZHU Dawei, LI Hailing, FENG Yali, SU Chaofan, XU Yina, LENG Haoyu, WANG Yongtao, ZHANG Yuemei, TAI Xiaochun, ZHANG Ying. Research Progress of Novel Vaccine Adjuvants [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2460-2467.
[2]	JIANG Haijun, WANG Dan, GUO Yu, WEI Li, HOU Lei, WANG Jing, ZHU Shanshan, LIU Jue. Fusion Expression and Immunogenicity of Recombinant Cap-flagellin Protein of Porcine Circovirus Type 3 [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 2044-2052.
[3]	LIU Xinyue, LI Lifang, WANG Zhenbao, ZHANG Wenliang, ZHANG Xin, ZHAO Hongtao, LI Pengyu, ZHENG Haotian, XIAO Jin, QI Peng. Adjuvants Screening and Immune Protection of Porcine Circovirus Type 2 Synthetic Peptide Vaccine [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1068-1080.
[4]	ZHOU Wenfeng, ZHANG Yingjie, BAI Yihan, ZHANG Luhua, MAI Jinhui, WANG Naidong, YANG Yi, ZHAN Yang, WANG Dongliang. Complete Genome Sequence Analysis and Cap CTL Epitopes Prediction of Porcine Circovirus Type 2 [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 656-665.
[5]	LIU Hongying, WANG Jingyu, CUI Zhenzhen, MA Qingxia, WANG Tingting, LI Jianming, SHI Kun, GONG Qinglong, DU Rui, LENG Xue. Research Progress on Immune Adjuvant Effect and Related Mechanism of Active Ingredients of Traditional Chinese Medicine on Animal Vaccines [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 390-397.
[6]	ZHAO Fengmiao, GUO Ting, ZHOU Yaping, ZHAO Hongmei, WANG Yuchen, TIAN Guangyuan, SUN Yajie, BIAN Yuchen, YU Jialiang, HAO Yongqing. Pathogenicity Analysis of Bovine Viral Diarrhea Virus and Immune Effect Evaluation of E2 Protein in New Zealand White Rabbits [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(9): 3549-3558.
[7]	ZHOU Wei, LIU Xuping, ZHANG Yanmin, PENG Wenjuan, ZHAO Liang, TAN Wensong. Serum-free Suspension Culture of PK-15 Cells and Its Effect on the Proliferation of Porcine Circovirus Type 2 [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(6): 2397-2405.
[8]	SHI Qian, XU Xinfeng, LI Xuewen, XIAO Yuanyuan, BAI Meihua, ZHANG Fuchun. Research Progress on the Mechanism of Polysaccharide as Adjuvant of Veterinary Vaccine [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(5): 1895-1907.
[9]	YANG Lulu, LI Anqi, YUAN Sheng, HUANG Liangzong, HUANG Shujian, WEN Feng, GUO Jinyue. Complete Genome Evolution Analysis of Four Porcine Circovirus Type 2 Isolates from Guangdong Province [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(3): 1015-1023.
[10]	SUN Qingshuai, YAN Weidong, LI Chang, WU Hao, LI Changchun, HE Qigai. Effect of PCV3 on the Expression of Growth-related Hormones and Functional Genes in Piglets [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(3): 1041-1047.
[11]	LAN Hong, REN Guangcai, YE Junxian, XIONG Ting, HE Xianming, LIU Yufu, YANG Zekun, HAO Wenqian, CHEN Ruiai. Isolation, Identification and Pathogenicity of Fowl Adenovirus Group Ⅰ Serotype 11 [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(3): 1067-1076.
[12]	ZHAO Yi, ZHANG Wen, CHEN Jiaji, SONG Manling, CHEN Qi, XIE Xiaodong, HU Tingjun. Establishment of 3D4/2 Cell Inflammatory Model Induced by PCV2 in vitro [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(12): 4920-4927.
[13]	SUN Qingshuai, CHEN Yanhong, YAO Lun, SUN Qi, YU Xuexiang, WU Hao, RUAN Shengnan, LEI Mingkai, YAN Weidong, HE Qigai. Diagnosis and Treatment of Reproductive Disorders in Sows Caused by Co-infection of PCV2 and PCV3 [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(10): 3982-3993.
[14]	PAN Chenhao, ZHANG Xin, ZHAO Ruili, JIANG Xuan, JIN Tianming, YU Enyuan, LI Liuan, ZENG Jun, YU Xiaoxue, SONG Qiqi, HU Ye. Construction and Cytotoxicity Detection of Recombinant Epitope Protein DCpep-GSE of Streptococcus suis [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(8): 2990-3001.
[15]	ZHU Weifeng, CHEN Lu, WANG Fang, HU Bo, RUAN Zhiyang, WANG Gaojie. Study on Immunoprotective Efficacy of Pasteurella Glycolytic Enzymes [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(6): 2188-2195.