不同免疫增强剂对禽戊型肝炎病毒ORF2蛋白的免疫效果分析  

Immune Effect of Different Immuno enhancers on ORF2 Protein of Avian Hepatitis E Virus

作　　者：闫洪健 高启明 王一新 常爽 赵鹏 王成森 YAN Hongjian;GAO Qiming;WANG Yixin;CHANG Shuang;ZHAO Peng;WANG Chengsen(College of Animal Science and Veterinary Medicine,Shandong Agricultural University,Tai′an,Shandong 271018;Shandong Vocational Animal Science and Veterinary College,Weifang,Shandong 261000)

机构地区：[1]山东农业大学动物科技学院(动物医学院),山东泰安271018 [2]山东畜牧兽医职业学院,山东潍坊261000

出　　处：《中国家禽》2023年第10期50-58,共9页China Poultry

基　　金：山东省重点研发计划(2022TZXD0019)。

摘　　要：为研究不同免疫增强剂对禽戊型肝炎病毒(aHEV)ORF2蛋白的免疫效果影响,研究通过大肠杆菌原核表达系统表达aHEV ORF2蛋白,纯化后与弗氏佐剂按1∶1等体积混合制备亚单位疫苗。将90只1日龄SPF雏鸡随机分成6组,每组15只。ORF2蛋白组单纯免疫YT-ORF2蛋白,ORF2蛋白+弗氏佐剂组免疫YT-ORF2蛋白+弗氏佐剂,AB101组免疫YT-ORF2蛋白+弗氏佐剂+AB101,AB106组免疫YT-ORF2蛋白+弗氏佐剂+AB106,AB506组免疫YT-ORF2蛋白+弗氏佐剂+AB506,空白对照组免疫生理盐水,分别在21、28、42日龄三次免疫SPF鸡。采用ELISA和间接免疫荧光检测不同免疫组SPF鸡的aHEV抗体水平,通过流式细胞仪检测CD4+和CD8+T淋巴细胞水平,荧光定量PCR检测IL-2、IL-4、IFN-γ3种细胞因子表达情况,并测定体重和器官发育指数。结果显示:原核表达获得分子量为72 ku的ORF2蛋白,AB106组产生的抗体水平极显著高于其他各组(P<0.01),AB106和AB506免疫增强剂可引起CD4^(+)和CD8^(+)T细胞比重显著增加并上调IL-2、IL-4、IFN-γ表达量(P<0.05),AB106组脾脏指数、法氏囊指数显著高于空白对照组、ORF2蛋白组和ORF2蛋白+弗氏佐剂组(P<0.05)。研究表明含胸腺肽和TLR7激动剂的免疫增强剂可显著提高CD4+和CD8+T细胞比重并上调与免疫相关的细胞因子表达量,其中且含胸腺肽的免疫增强剂诱导的aHEV抗体水平较高。In order to study the effect of different immunoenhancers on the immune effect of avian hepatitis E virus(aHEV)ORF2 protein,the expression of aHEV ORF2 protein was studied through E.coli prokaryotic expression system.After purifica⁃tion,the subunit vaccine was prepared by mixing it with Freund′s adjuvant in 1∶1 equal volume.Ninety 1-day-age SPF chickens were randomly divided into 6 groups with 15 chickens in each group.ORF2 protein group was immunized with YT-ORF2 protein alone,the ORF2 protein+Freund′s adjuvant group was immunized with YT-ORF2 protein+Freund′s adjuvant,the AB101 group was immunized with YT-ORF2 protein+Freund′s adjuvant+AB101 adjuvant,the AB106 group was immunized with YT-ORF2 protein+Freund′s adjuvant+AB106,the AB506 group was immunized with YT-ORF2 protein+Freund′s adjuvant+AB506,and the blank control group was immunized with physiological saline three times at 21,28 and 42 days of age,respectively.ELISA and indirect immunofluorescence were used to detect aHEV antibody levels in SPF chickens of different immune groups,CD4+and CD8^(+)T lymphocyte levels were detected by flow cytometry,and IL-2,IL-4 and IFN-γwere detected by fluorescence quanti⁃tative PCR expression of three cytokines,and body weight and organ development index were measured.The results showed that ORF2 protein with a molecular weight of 72 ku was obtained by prokaryotic expression.The antibody titer produced by chickens in AB106 group was significantly higher than that in other groups.AB106 and AB506 immunoenhancers could significantly in⁃crease the specific gravity of CD4+and CD8^(+)T cells and upregulate the expression of IL-2、IL-4、IFN-γ.The spleen and brusal index of SPF chickens in AB106 group was significantly higher than those in blank control group,ORF2 protein group and the ORF2 protein+Freund′s adjuvant group.It was indicated that immunoenhancers containing thymopeptide and TLR7 agonists could significantly increase the proportion of CD4+and CD8+T cells and up-regulate the expression of immu

关 键 词：禽戊型肝炎病毒 免疫增强剂 ORF2 亚单位疫苗 原核表达 

分 类 号：S855.3[农业科学—临床兽医学]