非洲猪瘟病毒重组载体疫苗激发/加强免疫策略的初步研究  

Prime/Boost Immunization Strategy of Vectored Vaccines against African Swine Fever Virus

作　　者：周晓慧 卢会鹏 张鑫宇[1] 夏晓莉[1] 孙怀昌[1] ZHOU Xiaohui;LU Huipeng;ZHANG Xinyu;XIA Xiaoli;SUN Huaichang(College of Veterinary Medicine,Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses,Yangzhou University,Yangzhou Jiangsu 225009,China;the Key Laboratory for High-Tech Research and Development of Veterinary Biopharmaceuticals,Jiangsu Agri-animal Husbandry Vocational College,Taizhou Jiangsu 225300,China)

机构地区：[1]扬州大学兽医学院,江苏省重要动物疫病与人兽共患病防控协同创新中心,江苏扬州225009 [2]江苏农牧职业技术学院,江苏泰州225300

出　　处：《西北农业学报》2023年第5期657-665,共9页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：国家重点研发计划(N2017YFD0502303,2018YFC0840404-3)。

摘　　要：为了选择合理的非洲猪瘟病毒(ASFV)重组载体的激发/加强免疫策略,构建表达ASFV CD2v-p30-p54融合抗原的重组伪狂犬病毒rPRV-F1,将其与表达相同融合抗原的重组腺病毒rAd-F1分为rAd-F1/rAd-F1、rAd-F1/rPRV-F1、rPRV-F1/rPRV-F1、rPRV-F1/rAd-F14个组合进行猪免疫试验,分别于免疫后不同时间采血并分离血清进行ELISA抗体检测,分离的外周血单个核细胞(PBMC)经ASFV抗原刺激后进行IFN-γ检测。免疫荧光与免疫转印试验结果显示,构建的rPRV-F1能在感染细胞中正确表达F1融合蛋白。所有免疫猪均能检测到抗原特异性抗体,加强免疫后的抗体水平显著升高。在测试的4个免疫方案中,尽管rAd-F1/rAd-F1组合初次免疫后的抗原特异性抗体水平和IFN-γ表达水平最高,但加强免疫后的抗体水平和IFN-γ表达水平最低。与此相反,尽管rPRV-F1/rAd-F1组合初次免疫后的抗体水平和IFN-γ表达水平最低,但加强免疫后的抗体水平和IFN-γ表达水平最高。这些研究结果表明rPRV/rAd是较好的ASFV重组载体疫苗的激发/加强免疫策略。To select a rational prime/boost strategy for immunization of vectored vaccine against African swine fever virus(ASFV),the recombinant pseudorabies virus which expressed ASFV CD2v-p30-p54 fusion antigen,namely rPRV-F1,was generated.Together with rAd-F1 expressing the same fusion antigen,four different prime-boost schemes rAd-F1/rAd-F1,rAd-F1/rPRV-F1,rPRV-F1/rPRV-F1,rPRV-F1/rAd-F1 used to immunize pigs and their serum samples were collected on different days for the antigen-specific antibody detection.Peripheral blood mononuclear cells(PBMCs)were isolated and stimulated with the three ASFV antigens for IFN-γdetection.Both immunofluorescence and Western blotting showed that the fusion antigen was correctly expressed in rPRV-F1-transduced cells.The antigen-specific antibodies were detected in all immunized pigs,which were significantly boosted by secondary immunization.Among the four immunization groups,the overall antibody level of rAd-F1/rAd-F1 immunization group was markedly increased after primary immunization,but it was the lowest after secondary immunization.In contrast,the overall antibody level of rPRV-F1/rAd-F1 immunization group was lower after primary immunization,but it was the highest after secondary immunization.Similarly,the overall antigen-specific IFN-γresponse of rAd-F1/rAd-F1 immunization group was the strongest after primary immunization,but it was the weakest after secondary immunization.In contrast,the overall IFN-γresponse of rPRV-F1/rAd-F1 immunization group was weaker after primary immunization,but it was the strongest after secondary immunization.These experimental data suggest that rPRV/rAd is the best prime/boost strategy for immunization of ASFV vectored vaccine.

关 键 词：非洲猪瘟病毒 重组载体疫苗 免疫策略 

分 类 号：S852.65[农业科学—基础兽医学]