Analysis of molecular bases determining tropism of negative strand viruses

决定负链病毒向性的分子碱基分析

基本信息

批准号：
15590411
负责人：
TAKEUCHI Kaoru
金额：
$ 2.3万
依托单位：
University of Tsukuba
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2004
项目状态：
已结题

项目摘要

The P gene of measles virus (MV) encodes the P protein and three accessory proteins (C,V and R). However, the role of these accessory proteins in the natural course of MV infection remains unclear. Here, we generated a recombinant wild-type MV lacking the C protein, wtMV(C-), using a reverse genetics system (Takeda et al., J.Virol.74:6643-6647). When 293 cells expressing the MV receptor SLAM (293/hSLAM) were infected with wtMV(C-) or parental wtMV, growth of wtMV(C-) was restricted, particularly in late stages. Consistently, EGFP-expressing wtMV(C-) induced late-stage cell rounding and cell death in the presence of the fusion-inhibiting peptide, suggesting that the C protein can prevent cell death and is required for long-term MV infection. Neutralizing antibodies against interferon-α/β did not restore the growth restriction of wtMV(C-) in the 293/hSLAM cells. When cynomolgus monkeys were infected with wtMV(C-) or wtMV, the number of MV-infected cells in the thymus was more than 1,000-fold lower for wtMV(C-) than wtMV. Immunohistochemical analyses showed strong expression of a MV antigen in the spleen, lymph nodes, tonsils and larynx of cynomolgus monkey infected with wtMV, but dramatically reduced expression in those tissues in cynomolgus monkey infected with wtMV(C-). These data indicate that the MV C protein is necessary for efficient MV replication both in vitro and in cynomolgus monkeys.

麻疹病毒（MV）的 P 基因编码 P 蛋白和三种辅助蛋白（C、V 和 R），然而，这些辅助蛋白在 MV 感染的自然过程中的作用仍不清楚。当 293 细胞表达 MV 时，使用反向遗传学系统（Takeda 等人，J.Virol.74：6643-6647）。受体 SLAM (293/hSLAM) 感染 wtMV(C-) 或亲本 wtMV，wtMV(C-) 的生长受到限制，特别是在晚期阶段，表达 EGFP 的 wtMV(C-) 诱导晚期细胞变圆。融合抑制肽存在时细胞死亡，表明 C 蛋白可以防止细胞死亡，并且是长期中和 MV 感染抗体所必需的。干扰素-α/β并不能恢复293/hSLAM细胞中wtMV(C-)的生长限制。当食蟹猴感染wtMV(C-)或wtMV时，胸腺中MV感染的细胞数量较多。 wtMV(C-) 比 wtMV 低 1,000 倍，免疫组织化学分析显示 MV 抗原在脾脏中强烈表达，感染wtMV的食蟹猴的淋巴结、扁桃体和喉部的表达显着降低，但感染wtMV(C-)的食蟹猴的这些组织中的表达显着降低。这些数据表明MV C蛋白对于MV在体外和体内的有效复制是必需的。食蟹猴。

项目成果

期刊论文数量（35）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Stringent requirement of the C protein of wild-type measles virus for growth both in vitro and in macaque

野生型麻疹病毒 C 蛋白在体外和猕猴体内生长的严格要求

DOI：
发表时间：
2005
期刊：
J Virol (In press)
影响因子：
0
作者：
Kadota S;Kanayama T;Miyajima N;Takeuchi K;Nagata K.;Kawaguchi A et al.;Takeuchi K et al.
通讯作者：
Takeuchi K et al.

Involvement of influenza virus PA subunit in assembly of functional RNA polymerase complexes