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Cloning, expression and antiviral activity of mink alpha-interferons

机译:貂α-干扰素的克隆,表达及抗病毒活性

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Background As a key link between innate and adaptive immune responses, the interferon (IFN) system is the first line of defense against viral infection. IFN, and in particular, IFN-α, has been used clinically as an effective therapeutic agent for viral infections. However, different subtypes of IFN-α demonstrate distinct antiviral activity. Therefore, it is important to identify IFN-α subtypes with high antiviral activity for the development of genetically engineered antiviral drugs. Results In this study, we cloned the genes for 13 IFN-α subtypes from peripheral blood lymphocytes of the mink. The homologies of the 13 mink IFN-α genes were 93.6–99.3% and 88.8–98.4% at the nucleotide and amino acid sequence levels, respectively. In contrast to human and canine IFN-α subtypes, most mink IFN-α subtypes contained two N-glycosylation sites. We expressed and purified 13 mink IFN-α subtypes in Escherichia coli . The cytopathic effect inhibition assay showed that all the 13 recombinant mink IFN-α subtypes inhibited the propagation of vesicular stomatitis virus in WISH cells, with IFN-α2 and IFN-α12 demonstrating the highest activities. Furthermore, recombinant mink IFN-α2 and IFN-α12 significantly suppressed the propagation of canine distemper virus in Vero cells, with IFN-α2 demonstrating the highest activity. Conclusions We identified the mink IFN-α2 subtype as a promising candidate for the development of effective antiviral drugs.
机译:背景技术作为先天性和适应性免疫反应之间的关键环节,干扰素(IFN)系统是抵抗病毒感染的第一道防线。 IFN,尤其是IFN-α,已经在临床上用作病毒感染的有效治疗剂。但是,IFN-α的不同亚型表现出独特的抗病毒活性。因此,鉴定具有高抗病毒活性的IFN-α亚型对于基因工程抗病毒药物的开发很重要。结果在这项研究中,我们从水貂的外周血淋巴细胞中克隆了13种IFN-α亚型的基因。 13个水貂IFN-α基因在核苷酸和氨基酸序列水平上的同源性分别为93.6–99.3%和88.8–98.4%。与人和犬的IFN-α亚型不同,大多数貂IFN-α亚型都含有两个N-糖基化位点。我们在大肠杆菌中表达和纯化了13种水貂IFN-α亚型。细胞病作用抑制试验表明,所有13种重组水貂IFN-α亚型均抑制WISH细胞中水泡性口炎病毒的传播,其中IFN-α2和IFN-α12表现出最高的活性。此外,重组水貂IFN-α2和IFN-α12显着抑制了犬瘟热病毒在Vero细胞中的繁殖,其中IFN-α2的活性最高。结论我们确定了水貂IFN-α2亚型是开发有效抗病毒药物的有希望的候选者。

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