生物矿化对新城疫病毒LaSota株生物学特性及免疫原性的影响

doi:10.11843/j.issn.0366-6964.2024.12.029

畜牧兽医学报 ›› 2024, Vol. 55 ›› Issue (12): 5663-5671.doi: 10.11843/j.issn.0366-6964.2024.12.029

生物矿化对新城疫病毒LaSota株生物学特性及免疫原性的影响

张高峰¹^,²(), 魏家阳², 冯贺龙²^,³, 李丽², 曾哲², 田光明¹, 聂仁锋⁴, 罗青平²^,⁵, 温国元²^,⁵, 魏红波¹^,*(), 商雨²^,*()

1. 长江大学动物科学技术学院，荆州 444100
2. 湖北省农业科学院畜牧兽医研究所，农业农村部畜禽细菌病防治制剂创制重点实验室/畜禽病原微生物学湖北重点实验室，武汉 430064
3. 山南市藏鸡产业研究院，山南 856000
4. 汉川市农业农村局，孝感 431600
5. 湖北洪山实验室，武汉 430070

收稿日期:2024-07-04 出版日期:2024-12-23 发布日期:2024-12-27
通讯作者: 魏红波,商雨 E-mail:zgf_9439@163.com;969788917@qq.com;suppershangyu@126.com
作者简介:张高峰(1999-)，男，湖北宜昌人，硕士，主要从事禽病原微生物相关研究，E-mail: zgf_9439@163.com
基金资助:
国家自然科学基金项目(U23A20239；32302869)；湖北省重点研发计划(2023BBB034)；国家现代农业产业技术体系资助(CARS-41)

Effects of Biomineralization on the Biological Characteristics and Immunogenicity of the LaSota Strain of Newcastle Disease Virus

ZHANG Gaofeng¹^,²(), WEI Jiayang², FENG Helong²^,³, LI Li², ZENG Zhe², TIAN Guangming¹, NIE Renfeng⁴, LUO Qingping²^,⁵, WEN Guoyuan²^,⁵, WEI Hongbo¹^,*(), SHANG Yu²^,*()

1. College of Animal Science and Technology, Yangtze University, Jingzhou 444100, China
2. Key Laboratory of Prevention and Control Agents for Animal Bacteriosis, Ministry of Agriculture and Rural Affairs / Hubei Provincial Key Laboratory of Animal Pathogenic Microbiology, Institute of Animal Husbandry and Veterinary, Hubei Academy of Agricultural Sciences, Wuhan 430064, China
3. Shannan Tibetan Chicken Industry Research Institute, Shannan 856000, China
4. Hanchuan Agriculture and Rural Bureau, Xiaogan 431600, China
5. Hongshan Laboratory, Wuhan 430070, China

Received:2024-07-04 Online:2024-12-23 Published:2024-12-27
Contact: WEI Hongbo, SHANG Yu E-mail:zgf_9439@163.com;969788917@qq.com;suppershangyu@126.com

摘要/Abstract

摘要：

旨在研究生物矿化对新城疫病毒(NDV)LaSota株生物学特性及免疫原性的影响，为耐热活疫苗研发提供新思路。通过测定不同矿化条件下病毒粒径、血凝效价和斑点印记，确定NDV的最佳生物矿化条件；将矿化后病毒接种BHK-21细胞，检测矿化对病毒增殖的影响，同时比较热处理后的矿化病毒在BHK-21细胞中的滴度变化情况；利用抗体中和试验检测矿化病毒的免疫原性；最后通过饮水免疫的方式评估矿化病毒在SPF雏鸡上的免疫效果。结果显示：生物矿化条件经优化后，LaSota株在4 mmol·L^-1 Na₂HPO₄和3 mmol·L^-1 CaCl₂条件下矿化效果最好，此时矿化病毒粒径最大，血凝效价最低，矿化率高达98.25%。矿化的LaSota株在BHK-21细胞上增殖起始时间迟于未矿化病毒，但两者的最终增殖滴度没有显著差异。经NDV抗体中和后，矿化病毒滴度的下降值为10^3.0 TCID₅₀·mL^-1，显著低于未矿化组。在56 ℃水浴中孵育15 min后，矿化的LaSota株病毒滴度仅下降10^3.5 TCID₅₀·mL^-1，与TS09-C耐热株的耐热特性相当。矿化的LaSota株免疫SPF雏鸡14 d后，抗体水平均高于未矿化LaSota株免疫鸡群。免疫后42和78 d进行攻毒试验，矿化LaSota株免疫鸡群的存活率分别为100%和60%。综上所述，本研究优化了NDV LaSota株的最佳矿化条件，矿化会延缓病毒的增殖速度，但不影响病毒的增殖滴度。经矿化后，能降低病毒与抗体的中和反应并提升其热稳定性，并可通过饮水免疫的方式对SPF雏鸡提供较好的保护效果。本研究证实通过生物矿化开发新城疫耐热活疫苗是一条切实可行的思路，也为开发其他病毒耐热活疫苗提供参考和借鉴。

关键词: 新城疫病毒LaSota株, 生物矿化, 生物学特性, 热稳定性, 免疫原性

Abstract:

This study aims to investigate the effects of biomineralization on the biological characteristics and immunogenicity of the Newcastle disease virus (NDV) LaSota strain, and provide new insights for the development of heat-stable live vaccines. The optimal biomineralization conditions for NDV were determined by measuring viral particle size, hemagglutination titers, and spot blots under different mineralization conditions. The effect of mineralization on viral replication was evaluated by inoculating mineralized viruses into BHK-21 cells, and then comparing their titer changes after heat treatment. The immunogenicity of mineralized viruses was evaluated through virus neutralization tests. Finally, the immunological efficacy of mineralized viruses was assessed in SPF chicks via water administration. Results were as follows: Upon optimization of the biomineralization conditions, the LaSota strain exhibited the best mineralization effect under 4 mmol·L^-1 Na₂HPO₄ and 3 mmol·L^-1 CaCl₂ conditions, characterized by the largest particle size of the mineralized virus, the lowest hemagglutination titer, and a high mineralization rate of 98.25%. Compared to the non-mineralized virus, the mineralized LaSota strain showed a delayed onset of replication in BHK-21 cells, however, there was no significant difference in the final replication titers between them. Following neutralization with NDV antibodies, the titer reduction value of the mineralized virus was 10^3.0 TCID₅₀·mL^-1, which was significantly lower than that of the non-mineralized group. After incubation at 56 ℃ for 15 minutes, the viral titer of the mineralized LaSota strain decreased by only 10^3.5 TCID₅₀·mL^-1, demonstrating a heat resistance characteristic comparable to that of the TS09-C heat-resistant strain. Fourteen days post-immunization, antibody levels in SPF chicks immunized with the mineralized LaSota strain were higher than those in chicks receiving the non-mineralized LaSota strain. Challenge experiments conducted 42 and 78 days post-immunization showed survival rates of 100% and 60%, respectively, in the group immunized with the mineralized LaSota strain. This study optimized the best biomineralization conditions for the NDV LaSota strain. Mineralization delays the initial viral replication but does not affect the final titer. After mineralization, it can reduce neutralization reaction of virus with antibodies and enhance thermal stability of virus, providing good protective effects to SPF chicks via drinking water immunization. This study confirms that developing heat-stable live vaccines for Newcastle disease through biomineralization is a practical and feasible approach, and also offers valuable references and inspirations for developing heat-stable live vaccines for other viruses.

Key words: LaSota strain of Newcastle disease virus, biomineralization, biological characteristics, thermal stability, immunogenicity

中图分类号:

S852.659.5

张高峰, 魏家阳, 冯贺龙, 李丽, 曾哲, 田光明, 聂仁锋, 罗青平, 温国元, 魏红波, 商雨. 生物矿化对新城疫病毒LaSota株生物学特性及免疫原性的影响[J]. 畜牧兽医学报, 2024, 55(12): 5663-5671.

ZHANG Gaofeng, WEI Jiayang, FENG Helong, LI Li, ZENG Zhe, TIAN Guangming, NIE Renfeng, LUO Qingping, WEN Guoyuan, WEI Hongbo, SHANG Yu. Effects of Biomineralization on the Biological Characteristics and Immunogenicity of the LaSota Strain of Newcastle Disease Virus[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(12): 5663-5671.

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免疫分组 Immunization group	雏鸡数量 Number of chicks	攻毒剂量 Challenge dose	免疫时间/d Days of immunity	死亡数 Number of deaths	存活数 Number of survivors	保护率/% Protection rate
矿化组1	5	10^4.5 EID₅₀	42	0	5	100
未矿化1	5	10^4.5 EID₅₀	42	0	5	100
对照组1	5	10^4.5 EID₅₀	42	5	0	0
矿化组2	5	10^4.5 EID₅₀	78	2	3	60
未矿化2	5	10^4.5 EID₅₀	78	4	1	20
对照组2	5	10^4.5 EID₅₀	78	5	0	0

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生物矿化对新城疫病毒LaSota株生物学特性及免疫原性的影响

Effects of Biomineralization on the Biological Characteristics and Immunogenicity of the LaSota Strain of Newcastle Disease Virus

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