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

作　　者：于晴晴 周祥瑜 李文昕 刘艳晶 王燕 和新文 何晨 邓国华[1,3] 施建忠 田国彬[1,3] 包红梅 曾显营[1,2,3] 陈化兰 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(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;Harbin Weike Biotechnology Company Limited,Harbin 150069;Key Laboratory for Prevention and Control of Avian Influenza and Other Major Poultry Diseases,Ministry of Agriculture and Rural Affairs,Harbin 150069)

机构地区：[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

出　　处：《中国农业科学》2025年第1期182-191,共10页Scientia Agricultura Sinica

基　　金：国家重点研发计划(2021YFD1800200);现代农业产业技术体系(CARS-41)。

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。【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 mine

关 键 词：禽流感 新城疫 多联多价 灭活疫苗 佐剂 免疫效果 

分 类 号：S859.797[农业科学—临床兽医学]