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目的制备人干扰素κ(hIFNκ)并初步研究其生物学活性。方法克隆hIFNκ的cDNA并根据大肠埃希菌的密码子偏好性人工合成了hIFNκ基因,在大肠埃希菌DH5α中高效表达、纯化后测定了rhIFNκ的多种生物学活性,包括抗病毒活性、抗细胞增殖活性、种属特异性以及NK细胞调节活性。结果成功地获得了高纯度的rhIFNκ,纯度在90%以上。利用WISHVSV系统检测rhIFNκ的抗病毒活力为2.0×106IU/mg。与rhIFNα2b的比较发现,rhIFNκ无论在抗病毒活性、细胞生长抑制,还是在促NK细胞活性方面均弱于前者。结论rhIFNκ在不同种属来源细胞上的抗病毒活性因种属来源而异,不同的病毒对rhIFNκ的敏感性是不同的。
Objective To prepare human interferon-κ (hIFNκ) and to study its biological activity. Methods The hIFNκ cDNA was cloned and the hIFNκ gene was synthesized based on the codon usage of Escherichia coli. The hIFNκ gene was highly expressed in Escherichia coli DH5α. After purification, various biological activities of rhIFNκ were assayed, including antiviral activity, anti- Cell proliferative activity, species-specific and NK cell regulatory activity. The results successfully obtained high purity rhIFNκ, purity above 90%. The antiviral activity of rhIFNκ detected by WISHVSV system was 2.0 × 106IU / mg. Comparisons with rhIFNα2b showed that rhIFNκ was weaker than the former both in antiviral activity, cell growth inhibition, and in promoting NK cell activity. Conclusion The antiviral activity of rhIFNκ on different species of cell originates from different genus sources, and the sensitivity of different viruses to rhIFNκ is different.