猪附红细胞体eno基因重组腺病毒疫苗对仔猪免疫效果的研究  

Study on the immune effects of recombinant Adenovirus vaccine of Mycoplasma suis eno gene on piglets

作　　者：杜宏鑫 韩金成 闫思雨 姜湛恒 张波 王金琦 薛书江[1] 崔起超 DU Hongxin;HAN Jincheng;YAN Siyu;JIANG Zhanheng;ZHANG Bo;WANG Jinqi;XUE Shujiang;CUI Qichao(College of Agriculture,Yanbian University,Yanji 133002,China;Laboratory Animal Center,Yanbian University,Yanji 133002,China)

机构地区：[1]延边大学农学院,吉林延吉133002 [2]延边大学实验动物中心,吉林延吉133002

出　　处：《黑龙江畜牧兽医》2024年第10期89-93,共5页Heilongjiang Animal Science And veterinary Medicine

基　　金：吉林省自然科学基金项目(YDZJ202201ZYTS616);吉林省教育厅“十三五”科学技术项目(JJKH20191134KJ)。

摘　　要：为了评价猪附红细胞体eno基因重组腺病毒疫苗(Ad5-M/eno重组腺病毒疫苗)对仔猪的免疫效果,试验将12头仔猪随机分为3组,分别为Ad5-M/eno重组腺病毒疫苗组、重组空载体腺病毒组和PBS对照组,分别于3组仔猪的颈部肌肉注射Ad5-M/eno重组腺病毒疫苗(浓度为1×10^(10)pfu/mL)、空载体重组腺病毒(浓度为1×10^(10)pfu/mL)和PBS各2.5 mL,于试验第0,21天分两次免疫。第2次免疫后第14天,麻醉后手术切除仔猪脾脏,采用密度梯度离心法提取猪脾脏淋巴细胞,利用流式细胞仪检测仔猪CD3^(+)、CD4^(+)和CD8^(+)T淋巴细胞含量。于第1次免疫前(第0天)及免疫后第7,14,21,28,35,42,49天无菌采集各组试验猪的颈静脉血,分离血清,检测每组仔猪抗猪附红细胞体特异性抗体IgG效价及IgG_(1)、IgG_(2a)和细胞因子白细胞介素-4(IL-4)、γ干扰素(IFN-γ)水平,评价Ad5-M/eno重组腺病毒疫苗对仔猪的细胞与体液免疫应答效果。结果表明:Ad5-M/eno重组腺病毒疫苗组的CD3^(+)、CD4^(+)和CD8^(+)T淋巴细胞含量极显著高于重组空载体腺病毒组和PBS对照组(P<0.01),Ad5-M/eno重组腺病毒疫苗组的CD4^(+)/CD8^(+)显著高于重组空载体腺病毒组和PBS对照组(P<0.05),重组空载体腺病毒组和PBS对照组之间的CD3^(+)、CD4^(+)和CD8^(+)T淋巴细胞含量及CD4^(+)/CD8^(+)差异不显著(P>0.05)。免疫后第28,35,42,49天,Ad5-M/eno重组腺病毒疫苗组的抗猪附红细胞体特异性IgG抗体效价及IgG_(1)、IgG_(2a)和细胞因子IL-4和IFN-γ水平极显著高于重组空载体腺病毒组与PBS对照组(P<0.01),重组空载体腺病毒组与PBS对照组之间差异不显著(P>0.05)。说明猪附红细胞体eno基因重组腺病毒疫苗可以诱导仔猪产生强烈的细胞免疫和体液免疫应答。In order to evaluate the immune effects of recombinant Adenovirus vaccine of Mycoplasma suis eno gene(Ad5-M/eno recombinant Adenovirus vaccine)on piglets,in this experiment,12 piglets were randomly divided into three groups,namely Ad5-M/eno recombinant Adenovirus vaccine group,recombinant empty vector Adenovirus group and PBS control group.Ad5-M/eno recombinant Adenovirus vaccine(1×10~(10)pfu/mL),empty vector recombinant Adenovirus(1×10~(10)pfu/mL)and PBS were injected intramuscularly into the neck muscles of the three groups of piglets,each for 2.5 mL.Immunization was given twice on days 0 and 21 of the test.Fourteen days after the second immunization,the piglet spleen was surgically removed after anesthesia;lymphocytes from porcine spleen were extracted by density gradient centrifugation.Flow cytometry was used to detect CD3~+,CD4~+and CD8~+T lymphocytes contents in piglets.The jugular venous blood of the test pigs was collected aseptically before the first immunization(day 0)and on the 7th,14th,21st,28th,35th,42nd,and 49th days after immunization;serum was isolated,and the levels of Mycoplasma suis-specific antibody IgG and IgG1,IgG2a,cytokine interleukin-4(IL-4)andγinterferon(IFN-γ)were detected in each group,for the evaluation of the effect of Ad5-M/eno recombinant Adenovirus vaccine on cellular and humoral immune responses in piglets.The results showed that the CD3~+,CD4~+and CD8~+T lymphocytes contents in the Ad5-M/eno recombinant Adenovirus vaccine group were significantly higher than those in the recombinant empty vector Adenovirus group and PBS control group(P<0.01).The CD4~+/CD8~+of the Ad5-M/eno recombinant Adenovirus vaccine group was significantly higher than that in the recombinant empty vector Adenovirus group and the PBS control group(P<0.05).There were no significant differences in CD3~+,CD4~+and CD8~+T lymphocytes contents and CD4~+/CD8~+between the recombinant empty vector Adenovirus group and the PBS control group(P>0.05).On the 28th,35th,42nd and 49th days after immunization,the levels o

关 键 词：猪附红细胞体 eno基因 重组腺病毒疫苗 仔猪 免疫保护 

分 类 号：S852.6[农业科学—基础兽医学]