Targeting Mechanism of a Novel Liver-targeting Interferon IFN-CSP Involves Liver Heparan Sulfate Proteoglycan

流式细胞术 蛋白多糖 干扰素 分子生物学 肝细胞 糖胺聚糖 化学 硫酸乙酰肝素 生物化学 生物 细胞外基质 免疫学 体外
作者
Xuemei Lu,Jie Wang,Xiaobao Jin,Yanting Huang,Wenting Zeng,Jiayong Zhu
出处
期刊:Current Drug Delivery [Bentham Science Publishers]
卷期号:13 (4): 528-533 被引量:4
标识
DOI:10.2174/1567201812666150827123602
摘要

In our previous study, a novel liver-targeting interferon (IFN-CSP) combining IFN α2b 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.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.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 IFNCSP binding to HepG2.2.15 cells and liver slices.These results indicate that the molecular mechanisms responsible for IFN-CSP targeting involve binding to HSPG of hepatocytes and liver.
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