猪脾转移因子对La Sota株鸡新城疫弱毒疫苗的免疫增强作用  

Immunoenhancement on Newcastle disease virus La Sota strain attenuated vaccine by pig spleen transfer factor

作　　者：徐磊[1] 杨慧[1] 刘毅发 杨子玚 闫丽萍 苏雅君 刘小龙 宋素泉[2] 张渊魁 黄瑜[5,6] XU Lei;YANG Hui;LIU Yifa;YANG Ziyang;YAN Liping;SU Yajun;LIU Xiaolong;SONG Suquan;ZHANG Yuankui;HUANG Yu(Fujian Vocational College of Agriculture,Fuzhou 350119,Fujian,China;College of Veterinary Medicine,Nanjing Agricultural University,Nanjing 210095,Jiangsu,China;Zhaofenghua Biotechnology(Nanjing)Co.,Ltd.,Nanjing 211102,Jiangsu,China;Present(Fuzhou)Biotech Company Limited,Fuzhou 350500,Fujian,China;Institute of Animal Husbandry and Veterinary Medicine,Fujian Academy of Agricultural Sciences,Fuzhou 350013,Fujian,China;Fujian Provincial Key Laboratory for Avian Diseases Control and Prevention,Fuzhou 350013,Fujian,China)

机构地区：[1]福建农业职业技术学院,福建福州350119 [2]南京农业大学动物医学院,江苏南京210095 [3]兆丰华生物科技(南京)有限公司,江苏南京211102 [4]派生特(福州)生物科技有限公司,福建福州350500 [5]福建省农业科学院畜牧兽医研究所,福建福州350013 [6]福建省禽病防治重点实验室,福建福州350013

出　　处：《微生物学报》2022年第2期727-741,共15页Acta Microbiologica Sinica

基　　金：福建省第七批引才“百人计划”;福建省科技计划项目(2019N0025,2021S2003);江苏省博士后科研资助计划(2019K072);福建省高层次人才和青年优秀人才访学研修资助计划(闽人社文〔2019〕239号);福州市科技计划项目(2020-PT-149);福建省畜禽疫病防控技术重大研发平台项目(2014N2003);福建省禽病防治重点实验室开放基金课题(KF201902);福建省家禽产业技术体系建设项目(2019-2022)。

摘　　要：【目的】本文旨在研究猪脾转移因子(TF)对新城疫(ND)病毒弱毒疫苗La Sota株的免疫增强作用及机理,为兽医临床防控提供理论依据。【方法】以4种不同剂量La Sota疫苗株分别单独、与TF联合免疫SPF鸡,14 d后以ND病毒(NDV)参考强毒F_(48)E_(9)株(10^(4.7) ELD_(50))进行攻毒,采用蛋白质芯片技术测定鸡外周血IL-6、IL-10、IL-16和IL-21浓度,并以血凝抑制(HI)试验和荧光定量RT-PCR方法分别检测鸡ND HI抗体效价和F_(48)E_(9)株的病毒血症水平。【结果】攻毒保护试验表明,单独免疫10^(5.17)、10^(4.17)、10^(3.17)和10^(2.17) EID_(50)剂量时疫苗攻毒保护率分别为100%、55%、0%和0%,半数保护量(PD_(50))为12023 EID_(50);对应剂量联合免疫时疫苗攻毒保护率分别为100%、75%、0%和0%,PD_(50)为6918 EID_(50);对照组(非免疫有攻毒)和空白组(非免疫非攻毒)死亡率分别为100%和0%。对10^(4.17) EID_(50)免疫剂量进一步检测分析表明,免疫后,联合免疫鸡IL-6、IL-10、IL-21和ND HI抗体效价(log_(2)x)比单独免疫最多时分别提高了254.95 pg/mL、62.10 pg/mL、1.51 pg/mL和2.6,其中ND HI抗体效价差异极显著(P<0.01);攻毒后,联合免疫鸡IL-10、IL-16、IL-21和病毒血症阳性率比单独免疫最多时分别降低了428.61 pg/mL、167.81 pg/mL、1.48 pg/mL和20%,其中IL-16、IL-21差异极显著(P<0.01)。【结论】本文结果显示,TF可提高La Sota株弱毒疫苗的免疫保护率及免疫后IL-6、IL-10、IL-21介导的免疫应答、ND HI抗体效价,降低疫苗PD_(50)及攻毒后IL-16、IL-21介导的炎症反应、IL-10介导的免疫应答抑制作用、病毒血症。本文明确了TF对La Sota株弱毒疫苗具有较好的免疫增强作用,为疫苗免疫佐剂研发和ND防控提供参考。[Objective]We evaluated the immunoenhancement effect and mechanism of pig spleen transfer factor(TF)on Newcastle disease virus(NDV)La Sota strain attenuated vaccine to provide a theoretical basis for the prevention and control of Newcastle disease(ND).[Methods]We inoculated La Sota strain alone and La Sota combined with TF into specific pathogen free(SPF)chickens at four different doses,and challenged SPF chickens by reference virulent NDV strain F_(48)E_(9)(10^(4.7) ELD_(50))14 days post vaccination.In addition,we determined the concentrations of IL-6,IL-10,IL-16,and IL-21 in chicken peripheral blood by protein chip technology,the hemagglutination inhibition(HI)antibody titer by HI assay,and the viremia level of F_(48)E_(9) by RT-PCR.[Results]The challenge test showed that the protection rates of vaccination with La Sota strain alone at the doses of 10^(5.17)、10^(4.17)、10^(3.17) and 10^(2.17) EID_(50) were 100%,55%,0%,and 0%,respectively,and the median protective dose(PD_(50))was 12023 EID_(50).The protection rates of vaccination with La Sota strain combined with TF at the corresponding doses were 100%,75%,0%,and 0%,respectively,and the PD_(50) was 6918 EID_(50).The mortality of control group(non-immunized but challenged)and blank group(non-immunized and non-challenged)were 100%and 0%.Upon vaccination at the dose of 10^(4.17) EID_(50),the IL-6,IL-10,IL-21,and ND HI antibody titer(log_(2)x)of the chicken exposed to combined vaccination increased at most by 254.95 pg/mL,62.10 pg/mL,1.51 pg/mL,and 2.6(P<0.01),respectively,compared with those in the chicken vaccinated with La Sota strain alone.After challenge,the IL-10,IL-16,IL-21,and the viremia levels in the chicken under combined vaccination decreased at most by 428.61 pg/mL,167.81 pg/mL(P<0.01),1.48 pg/mL(P<0.01),and 20%,respectively,compared with those in the chicken vaccinated with La Sota strain alone.[Conclusion]TF improves the immune protection rate of NDV attenuated La Sota strain vaccine,the immune response mediated by IL-6,IL-10,and IL-21,and the ND

关 键 词：猪脾转移因子 鸡新城疫弱毒La Sota株 鸡新城疫强毒F_(48)E_(9)株 免疫保护率 免疫增强作用 

分 类 号：S859.797[农业科学—临床兽医学]