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）都包含糖蛋白HA2亚基A螺旋区域中的共有序列。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％。由于肽似乎干扰了细胞中HA的适当折叠，因此使用战车将肽转染到细胞中，这能够有效地将肽递送到培养的细胞中，独立于内体途径。通过在病毒接种之前或之后输送肽来发现未发现抑制作用。这些数据可能表明，用于输送肽的内体途径在干扰HA的折叠方面非常有效，HA的折叠是在细胞内囊泡系统中进行的。为了为流感的未来流行病做准备，我们试图产生针对HA的H5和H2子类的单克隆抗体。在HA亚型HA的球头中发现了每种经过测试的每个病毒菌株的七个氨基酸残基，并引入与小鼠I -A -A^B MHC II类分子结合的框架成分。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。发现一条细胞系可产生针对A/新加坡/1/57（H2N2）流感病毒的中和抗体。尽管尚未研究与H5病毒的交叉反应性，但预计抗体将为未来流感的大流行提供治疗手段。