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

• 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
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15590411/
• 关键词: measles virus; reverse genetics; tropism; envelope protein; receptor; accessory protein; インターフェロン; 感染防御; シグナル伝達; 蛋白質リン酸化

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）P基因编码P蛋白和3个辅助蛋白（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。在此，我们使用反向遗传学系统产生了缺乏C蛋白的重组野生型MV，wtMV（C-）（Takeda等人，74：6643-6647）。当表达MV受体SLAM的293细胞（293/hSLAM）被wtMV（C-）或亲本wtMV感染时，wtMV（C-）的生长受到限制，特别是在后期。一致地，EGFP表达的wtMV（C-）诱导晚期细胞变圆和细胞死亡的融合抑制肽的存在下，这表明C蛋白可以防止细胞死亡，并需要长期MV感染。针对干扰素-α/β的中和抗体不能恢复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感染细胞的数量wtMV（C-）比wtMV低1，000倍以上。免疫组织化学分析显示MV抗原在感染wtMV的食蟹猴的脾、淋巴结、扁桃体和喉中强烈表达，但在感染wtMV的食蟹猴中这些组织中的表达显著降低（C-）。这些数据表明MV C蛋白对于体外和食蟹猴中的有效MV复制是必需的。

项目成果

Cell tropism of wild-type measles virus is affected by amino acid substitutions in the P.V and M proteins, or by a truncation in the C protein

野生型麻疹病毒的细胞向性受到 P.V 和 M 蛋白中氨基酸取代或 C 蛋白截短的影响

• 发表时间: 2004
• 期刊: Journal of General Virology 85(10)
• 作者: Miyajima;N. et al.
• 通讯作者: N. et al.

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

• 发表时间: 2005
• 期刊: J Virol (In press)
• 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

• DOI: 10.1128/jvi.79.2.732-744.2005
• 发表时间: 2005-01-01
• 期刊: JOURNAL OF VIROLOGY
• 影响因子: 5.4
• 作者: Kawaguchi, A;Naito, T;Nagata, K
• 通讯作者: Nagata, K

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

野生型麻疹病毒在体外和猕猴体内生长对 C 蛋白的严格要求

• 发表时间: 2005
• 期刊: J Virol (In press)
• 作者: Takeuchi K et al.

Wild-type measles virus induces large syncytium formation in primary human small airway epithelial cells by a SLAM (CD150)-independent mechanism

野生型麻疹病毒通过 SLAM (CD150) 独立机制诱导原代人小气道上皮细胞形成大合胞体

• 发表时间: 2003
• 期刊: Virus Res. 94
• 作者: Takeuchi K;Miyajima N;Nagata N;Takeda M;Tashiro M
• 通讯作者: Tashiro M