禽流感、新城疫二联五价灭活疫苗的免疫效果研究

doi:10.3864/j.issn.0578-1752.2025.01.014

中国农业科学 ›› 2025, Vol. 58 ›› Issue (1): 182-191.doi: 10.3864/j.issn.0578-1752.2025.01.014

禽流感、新城疫二联五价灭活疫苗的免疫效果研究

于晴晴¹(), 周祥瑜¹, 李文昕¹, 刘艳晶¹, 王燕¹, 和新文¹, 何晨¹, 邓国华¹^,³, 施建忠¹^,³, 田国彬¹^,³, 包红梅¹^,³, 曾显营¹^,²^,³^,^*(), 陈化兰¹^,³^,^*()

¹ 中国农业科学院哈尔滨兽医研究所/动物疫病防控全国重点实验室/国家禽类实验动物资源库，哈尔滨 150069
² 哈尔滨维科生物技术有限公司，哈尔滨 150069
³ 农业农村部禽流感等家禽重大疾病防控重点实验室，哈尔滨 150069

收稿日期:2024-08-04 接受日期:2024-11-08 出版日期:2025-01-01 发布日期:2025-01-07
通信作者:
曾显营，Tel：0451-51051678；E-mail：zengxianying@caas.cn。
陈化兰，Tel：0451-51997168；E-mail: chenhualan@caas.cn
联系方式: 于晴晴，Tel：0451-51051678；E-mail：1195446262@qq.com。
基金资助:
国家重点研发计划(2021YFD1800200); 现代农业产业技术体系(CARS-41)

Immune Effect Evaluation of Combined Inactivated Vaccine Against Avian Influenza and Newcastle Disease

YU QingQing¹(), ZHOU XiangYu¹, LI WenXin¹, LIU YanJing¹, WANG Yan¹, HE XinWen¹, HE Chen¹, DENG GuoHua¹^,³, SHI JianZhong¹^,³, TIAN GuoBin¹^,³, BAO HongMei¹^,³, ZENG XianYing¹^,²^,³^,^*(), CHEN HuaLan¹^,³^,^*()

¹ National Poultry Laboratory Animal Resource Center/State Key Lab for Animal Disease Control and Prevention/Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069
² Harbin Weike Biotechnology Company Limited, Harbin 150069
³ Key Laboratory for Prevention and Control of Avian Influenza and Other Major Poultry Diseases, Ministry of Agriculture and Rural Affairs, Harbin 150069

Received:2024-08-04 Accepted:2024-11-08 Published:2025-01-01 Online:2025-01-07

摘要/Abstract

摘要：

【背景】H5、H7、H9亚型禽流感病毒（avian influenza virus, AIV）和新城疫病毒（Newcastle disease virus, NDV）是严重危害禽业的重要病原，接种油乳剂型灭活疫苗是预防禽流感和新城疫的主要手段。当前养殖场预防这些传染病至少需要2种以上疫苗，导致家禽接种疫苗次数多，免疫负担重；另外，制备油乳剂型灭活疫苗的白油主要依赖进口，存在“卡脖子”风险。【目的】为了减少疫苗接种次数，降低免疫负担，对比国产和进口白油佐剂效果，本研究制备并评估了同时预防禽流感（H5+H7+H9）、新城疫的不同白油佐剂二联五价灭活疫苗，以期达到“一针防多病”的免疫效果，并为实现动物疫苗进口白油佐剂国产化替代提供数据支撑。【方法】将已构建的禽流感H5-Re13、H5-Re14、H7-Re4、H9-GX11583株和新城疫ND rLa-VII株共5株疫苗株分别接种鸡胚，收获尿囊液，经浓缩灭活后，按照1份抗原3份佐剂的比例加入不同的矿物油佐剂（Total、Marcol 52和HTM70），混合乳化，制成二联五价灭活疫苗，并分别以2 mL/只和0.5 mL/只的剂量通过肌肉注射接种3周龄SPF鸡，评价疫苗的安全性和有效性。【结果】免疫效力试验结果显示，3种白油佐剂制备的二联五价灭活疫苗对鸡均具有良好的安全性；3种白油佐剂疫苗接种鸡或免疫后均可产生针对H5-Re13株、H5-Re14株、H7-Re4株、H9-GX11583株和ND rLa-VII株有效的HI抗体，均可获得对效力检验毒株攻击的完全免疫保护。HI抗体持续期结果显示，3种白油佐剂灭活疫苗均可持续诱导高水平抗体半年以上，与进口白油（Total、Marcol 52）相比，国产白油HTM70诱导的HI抗体水平与进口白油Total差异不显著，高于进口白油Marcol 52，且国产白油佐剂HTM70批次间无明显差异。【结论】制备的禽流感（H5+H7+H9）、新城疫二联五价灭活疫苗对SPF鸡具有良好的免疫效果，且有望实现进口白油佐剂国产化替代。

关键词: 禽流感, 新城疫, 多联多价, 灭活疫苗, 佐剂, 免疫效果

Abstract:

【Background】 H5, H7 and H9 subtypes of avian influenza viruses (AIV) and Newcastle disease virus (NDV) are important pathogens that seriously jeopardize the avian industry, and vaccination with inactivated vaccines in the form of oil emulsions is the mainstay of prevention of avian influenza and Newcastle disease. At present, more than two vaccines are needed to prevent these infectious diseases in poultry farms, which leads to more times of vaccination and heavy immune burden on poultry. In addition, the mineral oil used to produce oil-emulsion inactivated vaccines is mainly dependent on imports, which has the risk of “neck sticking”. 【Objective】In order to reduce the number of vaccinations, lower the burden of immunization, and compare the effects of domestic and imported mineral oil adjuvants, this study developed and evaluated the inactivated vaccines, which could simultaneously prevent avian influenza (H5+H7+H9) and Newcastle disease by using different mineral oil adjuvants. This study expected to achieve the immunization effect of “one immunization preventing multiple diseases” and provide data support for the localization of mineral adjuvants for animal vaccines.【Method】A total of five vaccine strains of avian influenza H5-Re13, H5-Re14, H7-Re4, H9-GX11583 and Newcastle disease ND rLa-VII strains that have been constructed in the laboratory, which were inoculated into chicken embryos, harvested the allantoic fluid, concentrated and inactivated. Different mineral oil adjuvants (Total, Marcol 52 and HTM70) were added at a ratio of 1 part antigen to 3 parts of adjuvant. The mixture was then thoroughly mixed and emulsified to prepare the combined inactivated vaccine. Finally, the safety and efficacy of the vaccine were evaluated through intramuscular injection on three-week-old SPF chickens using doses of 2 mL and 0.5 mL, respectively.【Result】The results of the immunity efficacy test showed that the three kinds of mineral oil adjuvanted dichotomous multivalent inactivated vaccines had good safety for chickens; all three mineral oil adjuvanted vaccines could produce effective HI antibodies against H5-Re13, H5-Re14, H7-Re4, H9-GX11583 and ND rLa-VII strains after inoculation of chickens, and could obtain complete immune protection against the attack of the efficacy test strains. The results of HI antibody duration showed that all three mineral oil adjuvant inactivated vaccines could continuously induce high level of antibodies for more than half a year, compared with imported mineral oils (Total, Marcol 52), the level of HI antibody induced by domestic mineral oil HTM70 was close to that of imported mineral oil Total and higher than that of imported mineral oil Marcol 52, but the domestic mineral oil adjuvant HTM70 had no significant difference between batches.【Conclusion】The combined inactivated vaccine against avian influenza (H5+H7+H9) and Newcastle disease prepared in this study had a good immune efficacy on SPF chickens, and the domestic substitution of mineral oil adjuvant was expected.

Key words: avian influenza, Newcastle disease, multivalent, inactive vaccine, adjuvant, immune efficacy

于晴晴, 周祥瑜, 李文昕, 刘艳晶, 王燕, 和新文, 何晨, 邓国华, 施建忠, 田国彬, 包红梅, 曾显营, 陈化兰. 禽流感、新城疫二联五价灭活疫苗的免疫效果研究[J]. 中国农业科学, 2025, 58(1): 182-191.

YU QingQing, ZHOU XiangYu, LI WenXin, LIU YanJing, WANG Yan, HE XinWen, HE Chen, DENG GuoHua, SHI JianZhong, TIAN GuoBin, BAO HongMei, ZENG XianYing, CHEN HuaLan. Immune Effect Evaluation of Combined Inactivated Vaccine Against Avian Influenza and Newcastle Disease[J]. Scientia Agricultura Sinica, 2025, 58(1): 182-191.

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参考文献 32

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攻毒毒株 Challenge virus	组别（来源） Group（Source）	攻毒后第3天和第5天排毒阳性率排毒数/总数 Number of chickens shedding virus/Total chickens on days 3 and 5 post-challenge				保护率 Protection ratio
		第3天 Day 3		第5天 Day 5
		喉头 Oropharynx	泄殖腔 Cloaca	喉头 Oropharynx	泄殖腔 Cloaca
DK/FJ/S1424/2020（H5N6）	Total（进口Import）	0/10	0/10	0/10	0/10	100%
	Marcol 52（进口Import）	0/10	0/10	0/10	0/10	100%
	HTM70（国产Domestic）	0/10	0/10	0/10	0/10	100%
	对照组Control	10/10	10/10	*	*	0%
WS/SX/4-1/2020（H5N8）	Total（进口Import）	0/10	0/10	0/10	0/10	100%
	Marcol 52（进口Import）	0/10	0/10	0/10	0/10	100%
	HTM70（国产Domestic）	0/10	0/10	0/10	0/10	100%
	对照组Control	10/10	10/10	*	*	0%
CK/YN/SD024/2021（H7N9）	Total（进口Import）	0/10	0/10	0/10	0/10	100%
	Marcol 52（进口Import）	0/10	0/10	0/10	0/10	100%
	HTM70（国产Domestic）	0/10	0/10	0/10	0/10	100%
	对照组Control	10/10	10/10	*	*	0%
CK/GX/S11583/2019（H9N2）	Total（进口Import）	0/10	0/10	0/10	0/10	100%
	Marcol 52（进口Import）	0/10	0/10	0/10	0/10	100%
	HTM70（国产Domestic）	0/10	0/10	0/10	0/10	100%
	对照组Control	9/10	4/10	9/10	6/10	0%
CK/HeB/38/2009（NDV）	Total（进口Import）	0/10	0/10	0/10	0/10	100%
	Marcol 52（进口Import）	0/10	0/10	0/10	0/10	100%
	HTM70（国产Domestic）	0/10	0/10	0/10	0/10	100%
	对照组Control	10/10	10/10	*	*	0%

禽流感、新城疫二联五价灭活疫苗的免疫效果研究

Immune Effect Evaluation of Combined Inactivated Vaccine Against Avian Influenza and Newcastle Disease

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