Yuan Wu-Mei, Zhang Rui, Zhang Qian, Ma Fen-Lian, Wang Chao, Wang Ying-Zi, Zeng Yan, Zheng Li-Shu
Key Laboratory of Xinjiang Endemic and Ethnic Diseases, Department of Biochemistry, School of Medicine, Shihezi University, Shihezi, Xinjiang, China.
Key Laboratory for Medical Virology, National Health Commission, National Institute for Viral Disease Control and Prevention, China CDC, Beijing, China.
Protein Eng Des Sel. 2018 Sep 1;31(9):355-360. doi: 10.1093/protein/gzy029.
The previously generated recombinant human (rh) interferon (IFN)-λ1 protein has a short half-life, and this feature makes it challenging to conduct studies on potential clinical applications for rhIFN-λ1. In an attempt to overcome this difficulty, we constructed a 'long-life' version of rhIFN-λ1. This modified rhIFN-λ1, named rhIFN-λ1-CTPON, has a human chorionic gonadotropin β subunit carboxyl-terminal peptide (CTP) and an N-glycosylation sequence linked to its C-terminus. We confirmed the sequence of rhIFN-λ1-CTPON by mass spectrometry and then measured its biological activities. The results show that rhIFN-λ1-CTPON had antiviral activity and anti-proliferation activity in vitro that were similar to those of rhIFN-λ1 and that it similarly promoted natural killer cell cytotoxicity. Notably, the in vivo half-life of rhIFN-λ1-CTPON was determined to be 3-fold higher than that of rhIFN-λ1. We also assessed the anti-hepatitis B virus activity of rhIFN-λ1-CTPON; it was able to inhibit the production of the antigens HBs-Ag and HBe-Ag and induce antiviral gene expression. In conclusion, rhIFN-λ1-CTPON has a longer half-life than rhIFN-λ1 and has similar biological activities, so rhIFN-λ1-CTPON is an appropriate substitute for rhIFN-λ1 in the further study of potential clinical applications for rhIFN- λ1.
先前制备的重组人（rh）干扰素（IFN）-λ1蛋白半衰期较短，这一特性使得对rhIFN-λ1的潜在临床应用进行研究具有挑战性。为了克服这一困难，我们构建了一个“长寿命”版本的rhIFN-λ1。这种经过修饰的rhIFN-λ1，命名为rhIFN-λ1-CTPON，在其C末端连接了人绒毛膜促性腺激素β亚基羧基末端肽（CTP）和一个N-糖基化序列。我们通过质谱法确认了rhIFN-λ1-CTPON的序列，然后测定了其生物学活性。结果表明，rhIFN-λ1-CTPON在体外具有与rhIFN-λ1相似的抗病毒活性和抗增殖活性，并且同样促进自然杀伤细胞的细胞毒性。值得注意的是，rhIFN-λ1-CTPON的体内半衰期比rhIFN-λ1高3倍。我们还评估了rhIFN-λ1-CTPON的抗乙型肝炎病毒活性；它能够抑制抗原HBs-Ag和HBe-Ag的产生并诱导抗病毒基因表达。总之，rhIFN-λ1-CTPON比rhIFN-λ1具有更长的半衰期且具有相似的生物学活性，因此在对rhIFN-λ1潜在临床应用的进一步研究中，rhIFN-λ1-CTPON是rhIFN-λ1的合适替代品。
