目的:猪γ干扰素(poIFN-γ)和猪粒细胞-巨噬细胞集落刺激因子(poGM-CSF)是多功能细胞因子,对猪繁殖与呼吸综合征病毒(PRRSV)具有强大的抗病毒活性。在这项研究中,评价重组poIFN-γ-poGM-CSF融合蛋白在仔猪PRRSV灭活疫苗中的免疫佐剂作用。
方法:28只4周龄无特定病原体仔猪。
方法:实验仔猪分为对照组,高度病理性PRRSV,PRRSV灭病毒疫苗(KV),poIFN-γ-poGM-CSF,KV+1.0mgpoIFN-γ-poGM-CSF,KV+2.0mgpoIFN-γ-poGM-CSF,和KV+4.0mgpoIFN-γ-poGM-CSF组。通过重叠延伸PCR通过剪接构建重组poIFN-γ-接头-poGM-CSF融合基因,并使用大肠杆菌表达系统制备。之后评估其在PRRSVKV给药情况下的佐剂活性。
结果:该分析揭示了通过重叠延伸PCR通过剪接成功构建了poIFN-γ-接头-poGM-CSF融合基因,用重组poIFN-γ-接头-poGM-CSF在大肠杆菌中成功制备了用于表达具有肠激酶位点的硫氧还蛋白融合蛋白的质粒载体。重要的是,POIFN-γ-接头-POGM-CSF和PRRSVKV的共同给药显着增加中和抗体滴度,加速病毒清除,减少临床症状,预防高致病性PRRSV感染。
结论:重组poIFN-γ-poGM-CSF融合蛋白是用于猪免疫和病毒攻击的有希望的候选佐剂。
OBJECTIVE: Porcine interferon-γ (poIFN-γ) and porcine granulocyte-macrophage colony-stimulating factor (poGM-CSF) are multifunctional cytokines that exhibit robust antiviral activity against porcine reproductive and respiratory syndrome virus (PRRSV). In this study, the immunoadjuvant effects of recombinant poIFN-γ-poGM-CSF fusion protein in inactivated PRRSV vaccine administered to piglets were assessed.
METHODS: Twenty-eight 4-week-old specific pathogen-free piglets.
METHODS: The experimental piglets were divided into control, highly pathologic PRRSV, PRRSV killed virus vaccine (KV), poIFN-γ-poGM-CSF, KV + 1.0 mg poIFN-γ-poGM-CSF, KV + 2.0 mg poIFN-γ-poGM-CSF, and KV + 4.0 mg poIFN-γ-poGM-CSF groups. A recombinant poIFN-γ-linker-poGM-CSF fusion gene was constructed via splicing by overlap extension PCR and prepared using an Escherichia coli expression system, after which its adjuvant activity in the context of PRRSV KV administration was assessed.
RESULTS: This analysis revealed the successful construction of the poIFN-γ-linker-poGM-CSF fusion gene via splicing by overlap extension PCR, with recombinant poIFN-γ-linker-poGM-CSF successfully being prepared in E coli with a plasmid vector for expressing thioredoxin fusion proteins with an enterokinase site. Importantly, the coadministration of poIFN-γ-linker-poGM-CSF and PRRSV KV significantly increased neutralizing antibody titers, accelerated viral clearance, reduced clinical symptoms, and prevented highly pathogenic PRRSV infection.
CONCLUSIONS: The recombinant poIFN-γ-poGM-CSF fusion protein is a promising candidate adjuvant for use in the context of swine immunization and viral challenge.