College of Veterinary Medicine, Agricultural University of Hebei, Baoding, People's Republic of China.
Hebei Veterinary Biotechnology Innovation Center, Baoding, People's Republic of China.
Am J Vet Res. 2024 Jun 17;85(8). doi: 10.2460/ajvr.24.02.0025. Print 2024 Aug 1.
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.
Twenty-eight 4-week-old specific pathogen-free piglets.
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.
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.
The recombinant poIFN-γ-poGM-CSF fusion protein is a promising candidate adjuvant for use in the context of swine immunization and viral challenge.
猪干扰素-γ(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。本研究评估了重组猪干扰素-γ-猪粒细胞-巨噬细胞集落刺激因子融合蛋白在仔猪灭活 PRRSV 疫苗中的免疫佐剂作用。
28 头 4 周龄无特定病原体猪。
将实验仔猪分为对照组、高致病性 PRRSV 组、PRRSV 灭活病毒疫苗(KV)组、poIFN-γ-poGM-CSF 组、KV+1.0mg poIFN-γ-poGM-CSF 组、KV+2.0mg poIFN-γ-poGM-CSF 组和 KV+4.0mg poIFN-γ-poGM-CSF 组。通过重叠延伸 PCR 拼接构建了重组 poIFN-γ-接头-猪 GM-CSF 融合基因,并使用大肠杆菌表达系统进行了制备,然后评估了其在 PRRSV KV 给药情况下的佐剂活性。
本分析通过重叠延伸 PCR 拼接成功构建了 poIFN-γ-接头-猪 GM-CSF 融合基因,成功地在大肠杆菌中制备了带有肠激酶位点的硫氧还蛋白融合蛋白的质粒载体表达的重组 poIFN-γ-接头-猪 GM-CSF。重要的是,poIFN-γ-接头-猪 GM-CSF 与 PRRSV KV 的共同给药显著提高了中和抗体滴度,加速了病毒清除,减轻了临床症状,并预防了高致病性 PRRSV 感染。
重组 poIFN-γ-猪 GM-CSF 融合蛋白是一种有前途的候选佐剂,可用于猪免疫和病毒挑战。