Targeting Mechanism of a Novel Liver-targeting Interferon IFN-CSP Involves Liver Heparan Sulfate Proteoglycan
Targeting Mechanism of a Novel Liver-targeting Interferon IFN-CSP Involves Liver Heparan Sulfate Proteoglycan
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DOI：
10.2174/1567201812666150827123602
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发表时间：
2016-01-01
影响因子：
2.4
通讯作者：
Zhu, Jiayong
中科院分区：
文献类型：
--
作者：
Lu, Xuemei;Wang, Jie;Zhu, Jiayong
背景：在我们前期的研究中，利用大肠杆菌表达系统成功地设计并制备了一种新型的肝靶向干扰素(IFN-CSP)，该干扰素将干扰素α2b与疟原虫区I+肽结合在一起。纯化的干扰素-CSP具有抗乙肝病毒活性和肝靶向潜能。本研究旨在探讨干扰素-环磷酰胺在肝脏靶向性的分子机制。方法：采用免疫荧光染色方法检测干扰素-环磷酰胺在肝细胞中的结合部位。用激光共聚焦扫描显微镜观察肝组织中HSPG的分布与干扰素-CSP结合模式的对应关系。以肝细胞和肝组织为模型，用流式细胞仪和荧光显微镜观察了酶和可溶性糖胺聚糖对干扰素-CSP结合的影响。结果：肝细胞的研究表明，干扰素-CSP在肝细胞中的定位是质膜。对肝组织切片的研究表明，干扰素-CSP与肝组织结合的模式类似于硫酸乙酰肝素蛋白多糖(HSPG)免疫反应性的分布。肝素酶处理肝细胞和肝切片可降低干扰素-CSP与HepG2.2.15细胞和肝切片的结合。结论：干扰素-CSP靶向的分子机制可能与肝细胞和肝脏的HSPG结合有关。
Background: In our previous study, a novel liver-targeting interferon (IFN-CSP) combining IFN alpha 2b with plasmodium region I-plus peptide was successfully designed and prepared with Escherichia coli expression systems. The purified IFN-CSP showed anti-HBV activity and liver-targeting potentiality. The present investigation was designed to investigate the molecular mechanisms responsible for liver-targeting of IFN-CSP.Methods: The binding site of IFN-CSP in hepatocytes was assayed by immunofluorescent staining. The correspondence of HSPG distribution and the pattern of IFN-CSP binding in liver tissue were determined using a confocal laser scanning microscope. Both the hepatocytes and liver tissue were using as model to investigate the effect of enzyme and soluble glycosaminoglycan on IFN-CSP binding using flow cytometry and fluorescence microscope.Results: Studies of hepatocytes demonstrated that the localization of IFN-CSP in hepatocytes was the plasma membrane. Studies of liver tissue slices showed that IFN-CSP bound to liver tissue in a pattern similar to the distribution of heparan sulfate proteoglycan (HSPG) immunoreactivity. Pretreatment of hepatocytes and liver slices with heparinase reduced the binding of IFN-CSP to HepG2.2.15 cells and liver slices. Coincubation of IFN-CSP with heparin markedly inhibited IFN-CSP binding to HepG2.2.15 cells and liver slices.Conclusion: These results indicate that the molecular mechanisms responsible for IFN-CSP targeting involve binding to HSPG of hepatocytes and liver.