Recombinant human interleukin 28B: anti-HCV potency, receptor usage and restricted cell-type responsiveness
Recombinant human interleukin 28B: anti-HCV potency, receptor usage and restricted cell-type responsiveness
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重组人白细胞介素 28B：抗 HCV 效力、受体使用和受限细胞类型反应性
DOI：
10.1093/jac/dks015
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发表时间：
2012-05-01
影响因子：
5.2
通讯作者：
Yang, Wei
中科院分区：
文献类型：
--
作者：
Cheng, Min;Si, Youhui;Yang, Wei
白细胞介素28B （IL28B）遗传变异最近被报道为丙型肝炎病毒（HCV）对干扰素（IFN）治疗反应的有效预测因子。本研究的目的是在酵母中制备重组人il - 28b (rhIL28B)，并探讨其作为新型抗hcv药物的作用机制。建立了一种在毕赤酵母甲基营养酵母中生产rhIL28B的简单高效的方法。研究了rhIL28B的抗hcv活性、ifn刺激基因（ISGs）的诱导、受体使用和细胞反应性。通过一步离子交换纯化，可获得纯度约为95的可溶性rhIL28B。rhIL28B抑制HCV在Huh7.5.1细胞中的增殖，IC50为0.1510(3)mg/L。用rhIL28B处理肝癌细胞可在1 h内磷酸化STAT1并表达ISGs。rhIL28B的HCV抑制作用被受体IL10R2和IL28R1的抗体中和所拮抗。rhIL28B和利巴韦林联合使用可协同抑制细胞培养中HCV的产生。重要的是，与IFN-的广谱活性相比，我们证明了rhIL28B在肝、肺和前列腺细胞中的限制性细胞型反应性。本研究建立了一种简单、高效的方法，制备出具有较强的体外抗病毒活性和限制性细胞趋向性的rhIL28B，从而为改善hcv感染患者的治疗提供了一种新的抗病毒候选药物。
Interleukin 28B (IL28B) genetic variation has been recently reported as a potent predictor of hepatitis C virus (HCV) response to interferon (IFN) therapy. The aim of this study was to produce recombinant human IL28B (rhIL28B) in yeast and explore the action mechanisms of rhIL28B as a novel anti-HCV agent.A simple and efficient protocol for producing rhIL28B in the methylotrophic yeast Pichia pastoris was developed. The anti-HCV activity, induction of IFN-stimulated genes (ISGs), receptor usage and cellular responsiveness of rhIL28B were characterized.The yield of secreted rhIL28B was optimized to 200 mg/L, and soluble rhIL28B that was approximately 95 pure was achieved using a one-step ion-exchange purification procedure. rhIL28B inhibited HCV propagation in Huh7.5.1 cells with an IC50 of 0.1510(3) mg/L. Treatment of hepatoma cells with rhIL28B resulted in the phosphorylation of STAT1 within 1 h and expression of ISGs. The HCV inhibitory effects of rhIL28B were antagonized by the antibody neutralization of receptors IL10R2 and IL28R1. The combination of rhIL28B and ribavirin synergistically inhibited HCV production in cell culture. Importantly, compared with the broad-spectrum activity of IFN-, we demonstrated restricted cell-type responsiveness of rhIL28B in liver, lung and prostate cells.This study established an easy and highly efficient approach for the production of rhIL28B with potent in vitro antiviral activity and restricted cell tropism, and thus provides a novel antiviral candidate for improving the treatment of HCV-infected patients.