Tandem repeats of lactoferrin-derived anti-hepatitis C virus peptide enhance antiviral activity in cultured human hepatocytes
Tandem repeats of lactoferrin-derived anti-hepatitis C virus peptide enhance antiviral activity in cultured human hepatocytes
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DOI：
10.1111/j.1348-0421.2007.tb03882.x
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发表时间：
2007-01-01
影响因子：
2.6
通讯作者：
Kato, Nobuyuki
中科院分区：
文献类型：
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作者：
Abe, Ken-ichi;Nozaki, Akito;Kato, Nobuyuki
以前，我们发现牛乳铁蛋白（LF）和人乳铁蛋白（LF）在培养的非肿瘤性人肝细胞来源的PH 5CH 8细胞中特异性抑制丙型肝炎病毒（HCV）感染，并且我们鉴定了33个氨基酸残基（称为C-s3-33;氨基酸600-632）主要负责HCV E2包膜蛋白的结合活性和对HCV的抑制活性感染由于C-s3-33的抗HCV活性弱于人LF，我们推测E2蛋白结合活性的增加可能有助于抗HCV活性的增强。为了测试这种可能性，我们制备了C-s3-33的两个重复序列[（C-0-33）（2）]和三个重复序列[（C-s3 -33）（3）]并对其进行了表征。Far-Western印迹分析显示，（C-s3-33）和（C-ws 3 -33）（3）的E2蛋白结合活性比C-0-33的强，并且（C-s3-33）（3）的结合活性比（C-s3 - 33）（3）的强。使用PH 5CH 8细胞中的HCV感染系统，我们证明（C-s3-33）2和（C-s3-33）（3）的抗HCV活性比C-0-33更强。此外，使用最近开发的具有携带绿色荧光蛋白基因和天然E1和E2基因的VSV假型的感染系统，我们证明了（C-s3-33）（2）和（C-s3-33）（3）的抗病毒活性比C-s3-33强。这些结果表明，LF衍生的抗HCV肽的串联重复序列可用作抗HCV试剂。
Previously, we found that bovine and human lactoferrin (LF) specifically inhibited hepatitis C virus (HCV) infection in cultured non-neoplastic human hepatocyte-derived PH5CH8 cells, and we identified 33 amino acid residues (termed C-s3-33; amino acid 600-632) from human LF that were primarily responsible for the binding activity to the HCV E2 envelope protein and for the inhibiting activity against HCV infection. Since the anti-HCV activity of C-s3-33 was weaker than that of human LF, we speculated that an increase of E2 protein-binding activity might contribute to the enhancement of anti-HCV activity. To test this possibility, we made two repeats [(C-0-33)(2)] and three repeats [(C-s3-33)(3)] of C-s3-33 and characterized them. Far-Western blot analysis revealed that the E2 protein-binding activities of (C-s3-33), and (C-ws3-33)(3) became stronger than that of the C-0-33, and that the binding activity of (C-s3-33)(3) was stronger than that of (C-s3-33)(3). Using an HCV infection system in PH5CH8 cells, we demonstrated that the anti-HCV activities of (C-s3-33)2 and (C-s3-33)(3) became stronger than that of the C-0-33. Furthermore, using a recently developed infection system with a VSV pseudotype harboring the green fluorescent protein gene and the native E1 and E2 genes, we demonstrated that the antiviral activities of (C-s3-33)(2) and (C-s3-33)(3) were stronger than that of C-s3-33. These results suggest that tandem repeats of LF-derived anti-HCV peptide are useful as anti-HCV reagents.