摘要
制备猪圆环病毒2型(PCV2)多表位靶向性疫苗探讨其免疫应答类型及对仔猪免疫效力。用参考疫苗和PCV2多表位疫苗免疫21日龄PCV2和PRRSV抗原、抗体双阴性仔猪,流式细胞仪检测外周血T、B细胞含量和Th1/Th2比值,荧光定量RT-PCR检测IL-17、TGF-β及IFN-γ基因转录和ELISA测定IL-2、IL-4和IL-10蛋白表达,以期确定免疫应答类型。通过PCV2抗体效价和仔猪免疫攻毒保护试验评价疫苗免疫效力。结果显示,参考疫苗(A组)和PCV2多表位疫苗(B组)T细胞含量于免疫后7,14 d显著升高;Th1细胞在7~28 d显著升高,而Th2细胞含量无明显变化,使得Th1/Th2比值发生明显偏移,其中B组发生偏移程度较A组轻,表明疫苗免疫发生以Th1为主的T细胞免疫应答。A和B组IL-17和IFN-γ转录水平均显著升高,其中B组在7~14 d达到峰值,显著高于A组,显示B组细胞免疫活化和非特异性反应更强。外周血中IL-2蛋白含量A和B组均有所升高,组间差异不明显;IL-4蛋白含量A组在免疫后35 d明显高于B组,说明A组体液免疫强于B组。TGF-β基因转录水平和IL-10蛋白含量在免疫后无明显变化,显示均未导致免疫抑制的发生。A和B组疫苗均能刺激机体产生抗体,保护仔猪抵御PCV2强毒攻击,但B组疫苗免疫效力高于A组。结果表明,PCV2多表位靶向性疫苗主要刺激Th1介导的细胞免疫应答,激活IL-17和IFN-γ基因转录和IL-2、IL-4蛋白表达,有效保护免疫仔猪抵御PCV2强毒的攻击,具有良好的免疫保护效力。该研究为PCV2新型疫苗的研制和推广应用提供了技术和数据支撑。
The aim of this study was to determine the immune response type of PCV2 multi-epitope targeted vaccine and its immune efficacy to piglets.The 21 d piglets of the double negative PCV2 and PRRSV antigen and antibody were immunized with the vaccine in the study.The peripheral blood T,B lymphocyte content and Th1/Th2 content and ratio were measured by flow cytometry,and IL-17,TGF-βand IFN-γgene transcription levels were determined by fluorescence quantitative RT-PCR,and the protein expression of IL-2,IL-4 and IL-10 was determined by ELISA in the study.The immune efficacy of the vaccine was detected by the evaluation of PCV2 antibody titer and the immune protection test of piglets.The results showed that the content of T lymphocytes in the reference vaccine group(A group)and the PCV2 multi epitope vaccine group(B group)increased significantly at the 7 th and 14 th day.The results also showed that the Th1 cells increased significantly at day 7 and 28,but the content of Th2 cells did not change significantly,which resulted in a significant deviation of Th1/Th2 ratio.Compared with group A,the deviation of group B was less than that of group A,indicating that vaccine-induced T cell proliferation was stimulated by vaccine,which suggested that T cell immune response based on Th1 occurs in vaccine immunization.The results showed that the levels of IL-17 and IFN-γgene transcription increased significantly in both group A and B,and reached the peak at the 7 th day after immunization,which was significantly higher than that in group A,indicating that the ability of stimulating cell activation of IL-17 and IFN-γgene transcription in group B was stronger than that in group A.The content of IL-2 protein in peripheral blood was increased in group A and group B,but there was no significant difference between the two groups.The content of IL-4 protein in group A was significantly higher than that in group B at 35 days after immunization,indicating that humoral immunity in group A was stronger than that in group B.The transcriptio
作者
王艳萍
王金良
谢金文
徐倩倩
苗立中
董林
沈志强
WANG Yan-ping;WANG Jin-liang;XIE Jin-wen;XU Qian-qian;MIAO Li-zhong;DONG Lin;SHEN Zhi-qiang(Binzhou Animal Science and Veterinary Medicine Institute,Binzhou,Shandong 256600,China;Shandong Lvdu Bio-Sciences&Technology Co.Ltd,Binzhou,Shandong 256600,China;Shandong Binzhou Animal Propolis Vaccine Research and Development Promotion Center,Binzhou,Shandong 256600,China)
出处
《中国兽医学报》
CAS
CSCD
北大核心
2020年第6期1080-1086,1095,共8页
Chinese Journal of Veterinary Science
基金
山东省自然科学基金资助项目(ZR2017MC056)
山东省泰山产业领军人才工程高效生态农业创新类资助项目(LJNY2015014)。
