Studies on the behavior of hantavirus envelope glycoprotein in adsorption and invasion to target cells.

汉坦病毒包膜糖蛋白对靶细胞吸附和侵袭行为的研究。

基本信息

批准号：
17590410
负责人：
MORIMATSU Kumiko
金额：
$ 2.3万
依托单位：
HOKKAIDO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2006
项目状态：
已结题

项目摘要

Hantaviruses are causative agents for rodent-borne viral zoonosis, HFRS and HPS. Envelope glycoproteins G1 and G2 induce neutralizing antibodies. Monoclonal antibodies possessing neutralizing activities showed fusion inhibition activities. This means vaccine target epitope for hantavirus is a fusion responsible region in envelope protein. I studied the role of envelope glycoproteins in establishment of infection in cells. At the first, we found highly sensitive cell lines (A549, BHK and VeroE6 cells) and non-permissive cell lines (MDBK) by using pseudotype VSV screening. AlphaVbeta3 integrin was reported as candidate of the cellular receptor for hantavirus entry. Therefore, we tried to induce alphaVbeta3 into MDBK. I could not detect change of infectivity in that cells. However, transfection efficiency of plasmid constract to MDBK cells were not enough to evaluate the effect of molecules. Otherwise, anti-beta3 integrin antibodies were not inhibit cellular entry of hantavirus in permissive cells found in this study. So that this molecules was considered not to be major receptor molecules in these cells. Next, I screened bindings of glycolipids from permissive cell line Vero E6 cells against hantavirus glycoprotein. Any significant binding was not detected between glycolipids and envelope proteins. These results suggested that the cellular receptor of hantavirus must be protein molecules except alphaVbeta3 integrin. Finally, a clone 5 of MDBK cells was established as relative high DNA induction and non-permissive line. And cDNA libraly from permissive cell line A5449 was produced. This study established the basis for analysis of interaction between hantavirus envelope glycoproteins and cellular molecules.

汉坦病毒是啮齿动物传播的病毒性人畜共患病、HFRS 和 HPS 的病原体。包膜糖蛋白 G1 和 G2 诱导中和抗体。具有中和活性的单克隆抗体表现出融合抑制活性。这意味着汉坦病毒的疫苗靶标表位是包膜蛋白中的融合负责区域。我研究了包膜糖蛋白在细胞感染建立中的作用。首先，我们通过假型VSV筛选发现了高度敏感的细胞系（A549、BHK和VeroE6细胞）和非敏感细胞系（MDBK）。据报道，AlphaVbeta3 整合素是汉坦病毒进入细胞受体的候选者。因此，我们尝试将alphaVbeta3诱导进入MDBK。我无法检测到该细胞的感染性变化。然而，质粒构建体对MDBK细胞的转染效率不足以评估分子的效果。除此之外，抗β3整合素抗体不能抑制汉坦病毒进入本研究发现的允许细胞中。因此该分子被认为不是这些细胞中的主要受体分子。接下来，我筛选了来自许可细胞系 Vero E6 细胞的糖脂与汉坦病毒糖蛋白的结合。糖脂和包膜蛋白之间没有检测到任何显着的结合。这些结果表明，汉坦病毒的细胞受体一定是除αVβ3整合素之外的蛋白质分子。最后，建立MDBK细胞克隆5作为相对高DNA诱导和非许可系。并产生了来自许可细胞系A5449的cDNA文库。该研究为分析汉坦病毒包膜糖蛋白与细胞分子之间的相互作用奠定了基础。

项目成果

期刊论文数量（17）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Analysis of the immune response of Hantaan virus nucleocapsid protein-specific CD8+ T cells in mice.

DOI：
10.1016/j.virol.2007.02.039
发表时间：
2007-09
期刊：
Virology
影响因子：
3.7
作者：
Midori Taruishi;K. Yoshimatsu;K. Araki;M. Okumura;Ichiro Nakamura;K. Kajino;J. Arikawa
通讯作者：
Midori Taruishi;K. Yoshimatsu;K. Araki;M. Okumura;Ichiro Nakamura;K. Kajino;J. Arikawa

Arikawa J Development of serological assays for Thottapalayam virus, an insectivore-borne Hantavirus.

Arikawa J 开发了 Thottapalayam 病毒（一种食虫传播的汉坦病毒）的血清学检测方法。

DOI：
发表时间：
2007
期刊：
Clin Vaccine Immunol 14
影响因子：
0
作者：
Okumura M;Yoshimatsu K;Kumperasart S;Nakamura I;Ogino M;Taruishi M;Sungdee A;Pattamadilok S;Ibrahim IN;Erlina S;Agui T;Yanagihara R
通讯作者：
Yanagihara R

Development of serological assays for Thottapalayam virus, an insectivore-borne Hantavirus