Genetic Analysis and molecur Epidemiology of Parainfluenza virus type 4

4型副流感病毒的遗传分析及分子流行病学

• 批准号: 09670633
• 负责人: KOMADA Hiroshi
• 金额: $ 2.3万
• 依托单位: Suzuka University of Medical Science
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09670633/
• 关键词: Parainfluenza virus type 4; HN protein; Sequence; Incomplete replication; PCR; MK2 cell; L929 cell; typsin; パラインフルエンザ4型ウイルス; HNタンパク質; 点変異導入; フローサイトメトリー; 抗原性

cDNAs encoding human parainfluenza virus type 4B (hPIV-4B) haemagglutinin-neuraminidase (HN) protein were cloned and the nucleotide sequences were determined. A high degree of identity (81.4%) was observed between the nucleotide sequences of hPIV -4A and 4B HN proteins, and 87.3% identity was found between the deduced amino acid sequences. This degree of identity is considered to be greater than the immunological similarity between hPIV -4A and -4B HN proteins determined using mAbs. To elucidate the causes of the antigenic difference between HN proteins of hPIV-4A and -4B, we constructed three cDNAs of hPIV -4B HN whose potential N-glycosylation sites were partially or completely the same as in hPIV -4A HN cDNA. We compared the antigenicity of the expressed wild type and mutant proteins, and found that the antigenicities of the mutant hPIV -4B HN proteins were more similar to the hPIV -4A HN protein than to the non-mutant hPIN -4B HN protein. This study indicated that the antigenic div … More ersity between hPIV -4A and -4B was partly caused by deletion or creation of glycosylation sites, showing that the point mutations resulting in deletion or creation of glycosylation sites in one of the initial steps leading to the division of virus into subtypes.Human parainfluemza virus type 4A (hPIV -4A) and type 4B (hPIV -4B) were tested for their ability to replicate in the monkey kidney LLC-MK2 cell line (MK2 cells) and the murine L929 cell line (L929 cells). Both MK2 cells and L929 cell line(L929 cells). Both MK2 cells and L929 cells were non-permissive for replication of hPIV -4. Acetylated trypsin was effective for the virus replication in MK2 cells, but less effective in L929 cells. Interferon (IFN) produced endogeneously played no role in their replication in L929 cells. Synthesis of virus specific polypeptides was suppressed in L929 cells HN -mTNA was not detected. These results indicate that hPIV -4 can infect both MK2 cells and L929 cells. In MK2 cells, when protease exists in the extracellular medium, hPIV -4 exhibits multistep growth. In L929 cells, however, the cause of incomplete replication might be lack of other unknown factors. Less

克隆了编码人副流感病毒4 B型（hPIV-4 B）血凝素-神经氨酸酶（HN）蛋白的cDNA，并测定了其核苷酸序列。hPIV-4A和4 B HN蛋白核苷酸序列同源性较高（81.4%），推导的氨基酸序列同源性为87.3%。认为该同一性程度大于使用mAb测定的hPIV-4A和-4B HN蛋白之间的免疫学相似性。为了阐明hPIV-4A和-4B HN蛋白抗原性差异的原因，我们构建了3个与hPIV-4A HN cDNA具有部分或完全相同的N-糖基化位点的hPIV-4 B HN cDNA。比较了表达的野生型和突变型hPIV-4 B HN蛋白的抗原性，发现突变型hPIV-4 B HN蛋白的抗原性与hPIV-4A HN蛋白的相似性高于非突变型hPIN-4 B HN蛋白。这项研究表明，抗原区 ...更多信息 hPIV-4A和-4B之间的差异部分是由于糖基化位点的缺失或产生，显示在导致病毒分成亚型的初始步骤之一中导致糖基化位点缺失或产生的点突变。检测了hPIV-4A和4 B型（hPIV-4 B）在猴肾LLC-MK2细胞系（MK2细胞）和鼠L929细胞系（L929细胞）中复制的能力。MK2细胞和L929细胞系（L929细胞）。MK2细胞和L929细胞均不允许hPIV-4复制。乙酰化胰蛋白酶在MK 2细胞中对病毒复制有效，但在L929细胞中效果较差。内源性产生的干扰素（IFN）对它们在L929细胞中的复制没有作用。在L929细胞中病毒特异性多肽的合成受到抑制，HN-mTNA未检测到。这些结果表明，hPIV-4可以感染MK 2细胞和L929细胞。在MK 2细胞中，当细胞外培养基中存在蛋白酶时，hPIV-4表现出多步生长。而在L929细胞中，不完全复制的原因可能是缺乏其他未知因素。少

项目成果

Hiroshi Komada, Hisae Inoue, Chihiro Yamabayashi, Morihiro Ito, Mitsuo Kawano, Machiko Nishio, Masato Tsurudome, Yuji Kozuka, Naoya Noda, Kazuoshi Namba, Myles O'Brien and Yasuhiko Ito: "Incomplete replication of human parainfluenza virus type 4 in LLC-MK

Hiroshi Komada、Hisae Inoue、Chihiro Yamabayashi、Morihiro Ito、Mitsuo Kawano、Machiko Nishio、Masato Tsurudome、Yuji Kozuka、Naoya Noda、Kazuoshi Namba、Myles OBrien 和 Yasuhiko Ito：“人类副流感病毒 4 型在 LLC 中的不完全复制-

N.Tabata: "Protein tyrosine kinase activation provides an early and obligatory signalin anti-FRP-1/CD98/4F2 monoclonal antiboby induced cell fusion mediated byHIV gp160" Med.Microbiol.Immunol.186. 115-123 (1997)

N.Tabata：“蛋白酪氨酸激酶激活在抗 FRP-1/CD98/4F2 单克隆抗体诱导的 HIV gp160 介导的细胞融合中提供了早期和必需的信号”Med.Microbiol.Immunol.186。

Machiko Nishio, Masato Tsurudome, Morihiro Ito, Mitsuo Kawano, Shigeru Kusagawa, Hiroshi Komada and Yasuhiko Ito: "Mapping of domains on the human parainfluenza virus type 2 nucleocapsid protein (NP) required for NP-phosphoprotein or NP-NP interaction"Jou

Machiko Nishio、Masato Tsurudome、Morihiro Ito、Mitsuo Kawano、Shigeru Kusakawa、Hiroshi Komada 和 Yasuhiko Ito：“NP-磷蛋白或 NP-NP 相互作用所需的人类副流感病毒 2 型核衣壳蛋白 (NP) 上的结构域映射”Jou

Masato Tsurudome, Morihiro Ito, Shin-ichiro Takebayashi, Katsuzumi Okumura, Machiko Nishio, Mitsuo Kawano, Shigeru Kusagawa, Hiroshi Komada and Yasuhiko Ito: "Primary structure of the light chain of fusion reguratory protein-1/CD98/4F2 predicts a protein

Masato Tsurudome、Morihiro Ito、Shin-ichiro Takebayashi、Katsuzumi Okumura、Machiko Nishio、Mitsuo Kawano、Shigeru Kusakawa、Hiroshi Komada 和 Yasuhiko Ito：“融合调节蛋白-1/CD98/4F2 轻链的一级结构预测蛋白质

M.Ito: "Role of a single amino acid at the amino terminus of the simian virus 5F2 subunit in syncytium formation" Journal of Virology. 71. 9855-9858 (1997)

M.Ito：“猿猴病毒 5F2 亚基氨基末端单个氨基酸在合胞体形成中的作用”病毒学杂志。