Noor Aziz Ullah, Huipeng Lu, Du Zhanyu, Chengyi Song, Xiaohui Zhou, Xiaoming Liu, Khan Suliman, Sun Huaichang, Bellou Abdelouahab
Institute of Sciences in Emergency Medicine Department of Emergency Medicine, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, 106 Zhongshan Er Road, Guangzhou, 510080, Guangdong Province, China.
The College of Veterinary Medicine, Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Diseases and Zoonoses, Yangzhou University, Yangzhou, 225009, China.
Heliyon. 2024 Jul 19;10(14):e34725. doi: 10.1016/j.heliyon.2024.e34725. eCollection 2024 Jul 30.
Interferons play a major role in innate immunity and disease resistance. Porcine interferon alpha has 17 subtypes, and their gene sequences, tissue expression profiles, and antiviral activities have been primarily studied in domestic pigs but not in minipigs. Bama minipigs are genetically stable disease-resistant and making them as laboratory animal models for bioscience studies. To define the potential mechanism for disease resistance, in this study, we cloned 17 subtypes of Porcine interferon alpha genes in Bama minipigs using high fidelity polymerase chain reaction and subsequent sequencing. Sequence alignment showed that the 17 porcine interferon alpha subtypes were 98%-100 % homologous in those of domestic pigs. However, significantly different tissue expression profiles of PoIFN-α subtypes were found in the two pig species using real-time quantitative RT-PCR. Among the 10 different Bama minipig tissues tested, significant expression of multi-subtype porcine interferon alpha was detected in the lymph nodes and spleen, whereas no or low expression of fewer subtypes was detected in the heart, lung, brain, and small intestine. Sequence analysis revealed that the porcine interferon alpha promoters were almost similar between the two pig species. A cytopathic effect inhibition assay showed that the recombinant 17 porcine interferon alpha subtypes purified from mammalian cells had significantly different antiviral profile against vesicular stomatitis virus, porcine pseudorabies virus and porcine reproductive and respiratory syndrome virus compared with those in domestic pigs. Our findings provide evidence that porcine interferon alpha subtypes are highly conserved between Bama minipigs and domestic pigs but show varied tissue expression pattern and antiviral capabilities, which may contribute to their differences in disease resistance.
干扰素在先天免疫和抗病性中发挥着重要作用。猪α干扰素具有17个亚型,其基因序列、组织表达谱和抗病毒活性主要在家猪中进行了研究,但小型猪中尚未研究。巴马小型猪具有遗传稳定的抗病性,使其成为生物科学研究的实验动物模型。为了确定抗病性的潜在机制,在本研究中,我们使用高保真聚合酶链反应和后续测序克隆了巴马小型猪中的17种猪α干扰素基因亚型。序列比对显示,17种猪α干扰素亚型与家猪的同源性为98%-100%。然而,使用实时定量RT-PCR在两种猪中发现了猪干扰素-α亚型显著不同的组织表达谱。在测试的10种不同的巴马小型猪组织中,在淋巴结和脾脏中检测到多种亚型猪α干扰素的显著表达,而在心脏、肺、脑和小肠中未检测到或仅检测到较少亚型的低表达。序列分析表明,两种猪之间猪α干扰素启动子几乎相似。细胞病变效应抑制试验表明,从哺乳动物细胞中纯化的重组17种猪α干扰素亚型与家猪相比,对水泡性口炎病毒、猪伪狂犬病病毒和猪繁殖与呼吸综合征病毒具有显著不同的抗病毒谱。我们的研究结果提供了证据,表明猪α干扰素亚型在巴马小型猪和家猪之间高度保守,但显示出不同的组织表达模式和抗病毒能力,这可能导致它们在抗病性上的差异。