| 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 |
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| 引用本文：周晓慧，卢会鹏，张鑫宇，夏晓莉，孙怀昌.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[J].西北农业学报,2023,(5):657~665 |
| DOI:10.7606/j.issn.1004-1389.2023.05.001 |
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| 基金项目:国家重点研发计划（N2017YFD0502303，2018YFC0840404-3）。 |
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| 中文摘要: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-F1 4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。 |
| 中文关键词:非洲猪瘟病毒 重组载体疫苗 免疫策略 |
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| Prime/Boost Immunization Strategy of Vectored Vaccines against African Swine Fever Virus |
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| Abstract: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-γ detection. 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-γ response 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-γ response 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. |
| keywords:African swine fever virus Vectored vaccine Prime/boost immunization |
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