生物矿化的口蹄疫病毒样颗粒疫苗的免疫原性评价

doi:10.11843/j.issn.0366-6964.2023.04.023

畜牧兽医学报 ›› 2023, Vol. 54 ›› Issue (4): 1598-1607.doi: 10.11843/j.issn.0366-6964.2023.04.023

生物矿化的口蹄疫病毒样颗粒疫苗的免疫原性评价

李硕¹, 张韵¹, 白满元¹, 赵瑞翀¹, 宋河涛¹, 穆素雨¹, 滕志东¹, 董虎¹, 马娥宁¹, 孙世琪¹, 郭慧琛^1,2,3*, 尹双辉^1*

1. 中国农业科学院兰州兽医研究所, 家畜疫病病原生物学国家重点实验室/国家口蹄疫参考实验室, 兰州大学动物医学院, 兰州 730000;
2. 甘肃农业大学动物医学院, 兰州 730070;
3. 云南省畜牧兽医科学院云南热带亚热带动物病毒病实验室, 昆明 650224

收稿日期:2022-09-09 出版日期:2023-04-23 发布日期:2023-04-27
通讯作者: 郭慧琛,主要从事动物病毒病原生物学及以病毒为纳米生物材料的多学科交叉研究,Tel:+86-931-8312213;E-mail:guohuichen@caas.cn;尹双辉,主要从事动物疫苗与分子免疫学研究,Tel:0931-8341154;E-mail:yinshuanghui@caas.cn
作者简介:李硕(1997-),男,河南唐河人,硕士,主要从事口蹄疫疫苗评价研究,E-mail:ls199702@163.com
基金资助:
甘肃省科技重大专项（21ZD3NA001）；国家重点研发计划（2021YFD1800300）；国家自然科学基金（32072847；31873023；32072859；32002272）；科技人才与平台计划（202205AF150007）

Immunogenicity Evaluation of Biomineralized Foot-and-mouth Disease Virus-like Particles Vaccine

LI Shuo¹, ZHANG Yun¹, BAI Manyuan¹, ZHAO Ruichong¹, SONG Hetao¹, MU Suyu¹, TENG Zhidong¹, DONG Hu¹, MA E'ning¹, SUN Shiqi¹, GUO Huichen^1,2,3*, YIN Shuanghui^1*

1. State Key Laboratory of Veterinary Etiological Biology, College of Veterinary Medicine, Lanzhou University, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730000, China;
2. College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China;
3. Yunnan Tropical and Subtropical Animal Virus Diseases Laboratory, Yunnan Animal Science and Veterinary Institute, Kunming 650224, China

Received:2022-09-09 Online:2023-04-23 Published:2023-04-27

摘要/Abstract

摘要： 生物矿化技术可以增强口蹄疫（foot-and-mouth，FMD）病毒样颗粒（virus-like particles，VLPs）的耐热性，但并无试验数据支撑矿化FMD VLPs用于临床的可行性。本研究旨在评价矿化FMD VLPs疫苗在牛和猪上的免疫原性。用VLPs和矿化VLPs免疫猪，采用液相阻断ELISA （LPBE）检测猪血清中的特异性抗体滴度，并经酶联免疫斑点试验（ELISpot）检测免疫猪外周血淋巴细胞（PBLs）经体外刺激后分泌的特异性IFN-γ。用矿化A型VLPs、矿化O型VLPs、矿化的A型和O型二价VLPs分别免疫牛，采用LPBE、中和试验、夹心ELISA检测血清中的特异性抗体、中和抗体以及IL-4和IFN-γ的含量。结果显示，VLPs与矿化VLPs免疫猪的血清抗体水平无显著差异，相对于VLPs，矿化VLPs免疫猪的PBLs分泌的IFN-γ水平高于VLPs免疫猪（P<0.05）；3种疫苗免疫牛血清中IL-4和IFN-γ水平均高于PBS组，矿化二价疫苗与相应的矿化单价疫苗免疫牛的血清特异性抗体和中和抗体效价均无显著差异。综上所述，矿化VLPs对牛和猪均具有良好的免疫原性，不同血清型的矿化VLPs可以联合使用并且矿化壳层可增强VLPs诱导细胞免疫应答的能力。

关键词: 口蹄疫病毒, 病毒样颗粒疫苗, 生物矿化, 动物免疫

Abstract: Biomineralization technology can enhance the heat resistance of foot-and-mouth disease (FMD) virus-like particles (VLPs), but there is no experimental data to support the feasibility of mineralized FMD VLPs for clinical use. The aim of this study was to evaluate the immunogenicity of the mineralized FMD VLPs vaccine in cattle and pigs. The pigs were immunized with VLPs and mineralized VLPs, and the specific antibody titers in the serum of the immunized pigs were detected by liquid-phase blocking ELISA (LPBE). The specific IFN-γ secreted by peripheral blood lymphocytes (PBLs) after stimulation in vitro was detected by enzyme-linked immunospot assay (ELISpot). Cattle were immunized with mineralized type A VLPs, mineralized type O VLPs, and mineralized type A and type O bivalent VLPs, respectively. The serum specific antibody, neutralizing antibody, IL-4 and IFN-γ contents were detected by LPBE, neutralization test and sandwich ELISA. The results showed that there was no significant difference in serum antibody level between VLPs and mineralized VLPs immunized pigs.Compared with VLPs, the level of IFN-γ secreted by PBLs of mineralized VLPs immunized pigs was higher than that of VLPs immunized pigs (P<0.05). The serum of cattle immunized with the three vaccines contained more IL-4 and IFN-γ (P<0.05), and there were no significant differences in the titers of serum specific antibodies and neutralizing antibodies between the mineralized bivalent vaccine and the corresponding mineralized monovalent vaccine. In conclusion, mineralized VLPs have good immunogenicity for both cattle and pigs, mineralized VLPs of different serotypes can be used in combination, and mineralized shells can enhance the ability of VLPs to induce cellular immune response.

Key words: foot-and-mouth disease virus, virus-like particles vaccine, biomineralization, animal immunization

中图分类号:

S852.4

李硕, 张韵, 白满元, 赵瑞翀, 宋河涛, 穆素雨, 滕志东, 董虎, 马娥宁, 孙世琪, 郭慧琛, 尹双辉. 生物矿化的口蹄疫病毒样颗粒疫苗的免疫原性评价[J]. 畜牧兽医学报, 2023, 54(4): 1598-1607.

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.

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生物矿化的口蹄疫病毒样颗粒疫苗的免疫原性评价

Immunogenicity Evaluation of Biomineralized Foot-and-mouth Disease Virus-like Particles Vaccine

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