福建农业学报2025，Vol.40Issue(7)：669-677,9.DOI:10.19303/j.issn.1008-0384.2025.07.004
猪sPD-1蛋白体外增强猪圆环病毒Ⅱ型感染外周血单个核细胞的免疫效应
In Vitro Immunity Enhancement of Porcine Soluble PD-1 against PCV2 Infecting PBMCs in Pigs
摘要
Abstract
[Objective]Roles of the programmed cell death protein 1(PD-1)and its ligands(PD-Ls)in suppressing the immune response and the porcine soluble PD-1(sPD-1)protein in inhibiting the porcine circovirus type II(PCV2)infection on pigs were investigated in search for a new approach to control the disease in animals.[Method]The recombinant host,pET32a-PD1/Rosetta(DE),constructed in the laboratory was used for the extensive induction and purification to determine the sPD-1 expression conditions.Under which,a large amount of sPD-1 was purified to react with the peripheral blood mononuclear cells(PBMCs)infected by PCV2.The effects of PD-1 on the cells were measured by the indirect immunofluorescence and RT-qPCR,on the PBMC proliferation by CCK-8 detection and flow cytometry,and on the transcriptions and secretions of IL-2,IL-12,IL-21,IL-17A,and IFN-γ by RT-qPCR and ELISA.[Results]Highly purified recombinant sPD-1 showing high activity was successfully obtained.The viral load in the PBMCs induced with sPD-1 at 10 μg·mL-1 were significantly lower than that in those infected by PCV2 in the absence of sPD-1 to below 1 000 copies·μL-1.Meanwhile,the PBMCs proliferation index shown by CCK-8 significantly increased(P＜0.05).The flow cytometric measurement on fluorescence intensity of(68.6±10.14)%on the PBMCs was significantly higher with the presence of sPD-1 than(28.7±3.18)%on the counterparts without the soluble protein(P＜0.05).The RT-qPCR and ELISA also showed the cytokine transcriptions and secretions of IL-2,IL-12,and IFN-γ to be significantly higher under the same condition(P＜0.05).[Conclusion]The presentation of porcine sPD-1 significantly inhibited PCV2 infection on PBMCs through the related metabolic pathways.At the same time,it promoted the PBMC cell proliferation,cytokine transcriptions,and secretion.Therefore,the protein could potentially be used in mitigating PCV2 infection on animals.关键词
猪sPD-1蛋白/猪圆环病毒/外周血单个核细胞/体外/免疫反应Key words
soluble porcine PD-1 protein/PCV2/PBMCs/in vitro/immunoregulation分类
农业科技引用本文复制引用
何勇,张心杰,朱艳平,岳锋,姚倩文,吴玉苹,周娟娟..猪sPD-1蛋白体外增强猪圆环病毒Ⅱ型感染外周血单个核细胞的免疫效应[J].福建农业学报,2025,40(7):669-677,9.基金项目
河南省高等学校重点科研项目(22A230017) （22A230017）
河南省重点研发专项(241111112900) （241111112900）
新乡市软科学资助计划项目(RKX2024012) （RKX2024012）
河南省大学生创新创业训练计划项目(20211071019、202411071012) （20211071019、202411071012）
