Development of antiviral drugs inhibiting fusion between viral envelope and cellular membrane

开发抑制病毒包膜与细胞膜融合的抗病毒药物

基本信息

批准号：
18590453
负责人：
OKAZAKI Katsunori
金额：
$ 2.53万
依托单位：
Health Sciences University of Hokkaido
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

项目摘要

We found that each subtype (H1-H16) of influenza virus hemagglutinin (HA) contained the consensus sequence in a-helix region of HA2 subunit of the glycoprotein. Virus replication in MDCK cells was reduced by 10% when synthetic peptides with this sequence were added in the medium. Since the peptides seemed to interfere with proper folding of HA in the cells, the peptides were transfected into the cells using Chariot, which is capable of efficiently delivering peptide into cultured cells independently of the endosomal pathway. No inhibition was found by delivering the peptides before or after virus inoculation. These data may indicate that the endosomal pathway for delivering the peptides is much effective in the interference of the folding of HA, which is carried out in the intracellular vesicle system.To prepare for the future pandemic of influenza, we attempted to produce monoclonal antibodies against H5 and H2 subtypes of HA. Seven amino acid residues common to each virus strain tested were found in the globular head of HA of both HA subtypes and introduced into the frame component bound to mouse I -A^b MHC class II molecule. The splenocytes from C57BL/6 mice immunized with the synthetic peptides containing the seven residues were used to prepare hybridoma cells. One cell line was found to produce neutralizing antibodies against A/Singapore/1/57 (H2N2) influenza virus. Although cross-reactivity with H5 virus of the antibodies is not yet studied, it is expected that the antibodies would provide therapeutic means for future pandemic of influenza.

我们发现流感病毒血凝素（HA）的每个亚型（H1-H16）在糖蛋白HA2亚基的α螺旋区域都含有共有序列。当在培养基中添加具有该序列的合成肽时，MDCK 细胞中的病毒复制减少了 10%。由于这些肽似乎会干扰细胞中 HA 的正确折叠，因此使用 Chariot 将这些肽转染到细胞中，Chariot 能够独立于内体途径将肽有效地递送到培养细胞中。在病毒接种之前或之后递送肽没有发现抑制作用。这些数据可能表明，递送肽的内体途径对于干扰HA折叠非常有效，HA折叠是在细胞内囊泡系统中进行的。为了为未来的流感大流行做好准备，我们尝试生产针对HA的单克隆抗体。 HA 的 H5 和 H2 亚型。在两种HA亚型的HA的球状头部中发现了每个测试病毒株共有的7个氨基酸残基，并将其引入到与小鼠I-A^b MHC II类分子结合的框架组件中。用含有七个残基的合成肽免疫的C57BL/6小鼠的脾细胞用于制备杂交瘤细胞。发现一种细胞系可产生针对 A/Singapore/1/57 (H2N2) 流感病毒的中和抗体。虽然该抗体与H5病毒的交叉反应性尚未研究，但预计该抗体将为未来流感大流行提供治疗手段。