[1]李玲,祝博森,朱婷,等.双靶向融合蛋白用于新城疫病毒La Sota株对鸭的免疫效率的提高作用[J].江苏农业学报,2024,(12):2317-2323.[doi:doi:10.3969/j.issn.1000-4440.2024.12.015]
　LI Ling,ZHU Bosen,ZHU Ting,et al.Immunoenhancement of Newcastle disease virus La Sota strain in ducks by dual-targeting fusion protein[J].,2024,(12):2317-2323.[doi:doi:10.3969/j.issn.1000-4440.2024.12.015]
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双靶向融合蛋白用于新城疫病毒La Sota株对鸭的免疫效率的提高作用()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2024年12期
页码:
2317-2323
栏目:
畜牧兽医·水产养殖·益虫饲养
出版日期:
2024-12-30

文章信息/Info

Title:
Immunoenhancement of Newcastle disease virus La Sota strain in ducks by dual-targeting fusion protein
作者:
李玲1; 祝博森1; 朱婷1; 商璐2; 3; 邓碧华3; 4; 卢宇3; 4; 徐海2; 3
(1.江苏农牧科技职业学院,江苏泰州225300；2.江苏省农业科学院泰州农科所,江苏泰州225300；3.兽用生物制品<泰州>国泰技术创新中心,江苏泰州225300；4.江苏省农业科学院动物免疫工程研究所,江苏南京210014)
Author(s):
LI Ling1; ZHU Bosen1; ZHU Ting1; SHANG Lu2; 3; DENG Bihua3; 4; LU Yu3; 4; XU Hai2; 3
(1.Jiangsu Agri-animal Husbandry Vocational College, Taizhou 225300, China;2.Taizhou Institute of Agricultural Sciences, Jiangsu Academy of Agricultural Sciences, Taizhou 225300, China;3.GuoTai (Taizhou) Center of Technology Innovation for Veterinary Biologicals, Taizhou 225300, China;4.Institute of Veterinary Immunology ＆ Engineering, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China)
关键词:
新城疫病毒; 融合蛋白; 靶向; 免疫效率
Keywords:
Newcastle disease virus; fusion protein; targeting; immune efficiency
分类号:
S831.7
DOI:
doi:10.3969/j.issn.1000-4440.2024.12.015
文献标志码:
A
摘要:
本研究旨在探究双靶向融合蛋白用于新城疫病毒(NDV) La Sota毒株对鸭的免疫效率的提高作用,为提升鸭新城疫免疫防控能力做有益探索。将特异性结合囊膜病毒表面糖蛋白的红藻凝集素(GRFT)与靶向结合禽树突状细胞的纳米抗体分子(VHH)的编码基因进行串联表达,经大肠杆菌表达系统制备双靶向融合蛋白。利用组氨酸(His)标签纯化GRFT-VHH融合蛋白,采用有限稀释法定量评价融合蛋白与La Sota病毒的结合能力。用饱和结合融合蛋白的La Sota病毒制备疫苗,分组免疫无特定病原体（SPF）雏鸭,监测免疫后血凝抑制(HI)抗体效价以及IL-4、IFN-γ细胞因子水平。结果显示,构建的重组大肠杆菌能够高效表达融合蛋白,经Ni柱纯化回收的GRFT-VHH54、GRFT-VHH74蛋白质量浓度分别为230 μg/mL、1 350 μg/mL,仅需100 ng融合蛋白即可完全结合1×108半数感染量（EID50）病毒；NDV+GRFT-VHH54组免疫后21 d HI抗体效价达到7 log22以上,血清中IL-4和IFN-γ含量分别为80.0 pg/mL、8.8 pg/mL,显著高于NDV单独免疫组(P<0.05)。双靶向融合蛋白GRFT-VHH54能够提高新城疫病毒La Sota株对鸭的免疫效率,可作为免疫增强剂做进一步的开发与应用。
Abstract:
This study aimed to explore the enhanced immunogenicity of the Newcastle disease virus (NDV) La Sota strain in ducks through a dual-targeting fusion protein, providing valuable insights for improving NDV immune prevention and control capabilities. The genes encoding Griffithsin (GRFT) that specifically binds to the surface glycoprotein of enveloped viruses and nanoantibody molecules (VHH) targeting avian dendritic cells were co-expressed in tandem using the Escherichia coli system and purified via a His-tag. The binding capacity of the GRFT-VHH fusion protein to the La Sota virus was quantitatively assessed using a limited dilution method. Vaccines were prepared with La Sota virus saturated with the fusion protein and administered to groups of specific pathogen free (SPF) ducklings. After immunization, the hemagglutination inhibition (HI) antibody titer and levels of cytokines IL-4 and IFN-γ were evaluated. Results indicated that the constructed recombinant E. coli efficiently expressed the fusion protein. The concentrations of GRFT-VHH54 and GRFT-VHH74 purified by Ni column were 230 μg/mL and 1 350 μg/mL, respectively. Only 100 ng of the fusion protein was required to fully bind 1×108 median infective dose (EID50) virus. The HI antibody titer in the NDV+GRFT-VHH54 group reached more than 7 log22, and the levels of IL-4 and IFN-γ in serum were 80.0 pg/mL and 8.8 pg/mL respectively at 21 d post-immunization, which were significantly higher than those in the NDV group (P<0.05). The dual-targeting fusion protein GRFT-VHH54 can enhance the immunogenicity of the NDV La Sota strain in ducks, and can be used as an immunopotentiator for further development and application.

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备注/Memo

备注/Memo:
收稿日期:2024-06-30基金项目:江苏农牧科技职业学院院级课题(NSF2023ZR13)作者简介:李玲(1981-),女,江苏盐城人,硕士,讲师,主要从事动物疾病防控研究。(E-mail)ll20220215@163.com通讯作者:徐海, (E-mail)hai_x@126.com
更新日期/Last Update: 2025-01-23