Studies on a mechanism of hantavirus persistent infection in central nervous system and immune system

汉坦病毒持续感染中枢神经系统和免疫系统机制研究

基本信息

批准号：
10306019
负责人：
ARIKAWA Jiro
金额：
$ 22.78万
依托单位：
Hokkaido University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A).
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 2000
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10306019/
关键词：
Hemorrhagic fever with renal syndrome HFRS hantavirus zoonosis virus infection pathogenicity genetic reassortant infection enhancement

项目摘要

1. Hantavirus infection was enhanced about 10 times by addition of lectins (DBA and SBA) which specifically bind to N-acetylgalactosamine. This enhancement was considered to caused by the cross linking between receptor to virion by the lectins.2. Two Vero E6 cell subclones, one induce low pH dependent cell fusion after hantavirus infection and the other resistant to cause cell fusion, were established. By using the two cell clones, the possible role for the cellular factor for responsible for induce infected cell fusion was considered.3. After peripheral infection of hantavirus to mice, virulent virus reached to brain 4 to 5 days earlier than that of avirulent virus. This difference was considered as a mechanism which define the virulence of hantavirus in the mouse model.4. Virulent hantavirus showed higher growth rate in the brain micro vascular cells and peritoneal macrophage, suggesting that virulence relates to the growth ability in the target organs.5. Genetic reassortant virus between virulent and avirulent viruses were established. The comparison of the virulence of the reassortant viruses indicated that one amino acid difference at enveloped protein and nucleotide difference in the polymerase gene are related to the virulence.6. Virulent hantavirus infected SCID mice were passively transferred with spleen cells of immunized BALB/c mice 3 weeks after infection. The recipient SCID mice represented apparent weight loss 4 days after the transfer, indicating the immune mediated pathogenicity.7. Hantaan virus S and M genome segment which encoding nucleocapsid protein and enveloped glycoprotein, respectively were cloned and expressed in the mammalian cells.8. Entire L genome segment which encodes polymerase was cloned. The expression of the L clone is under going.

1. 通过添加与N-乙酰半乳糖胺特异性结合的凝集素（DBA和SBA），汉坦病毒感染增强约10倍。这种增强被认为是由凝集素在受体与病毒体之间的交联引起的。2．建立了两种 Vero E6 细胞亚克隆，一种在汉坦病毒感染后诱导低 pH 依赖性细胞融合，另一种抵抗引起细胞融合。利用这两个细胞克隆，探讨了诱导感染细胞融合的细胞因子的可能作用。 3．小鼠外周感染汉坦病毒后，强毒病毒比无毒病毒早4～5天到达脑部。这种差异被认为是定义汉坦病毒在小鼠模型中毒力的机制。 4．强毒汉坦病毒在脑微血管细胞和腹腔巨噬细胞中表现出较高的生长速度，表明毒力与靶器官的生长能力有关。 5．建立了强毒力和无毒力病毒之间的基因重配病毒。重配病毒毒力比较表明，包膜蛋白的1个氨基酸差异和聚合酶基因的核苷酸差异与毒力有关。 6．感染强毒汉坦病毒的 SCID 小鼠在感染后 3 周被动转移免疫 BALB/c 小鼠的脾细胞。移植后4天，SCID受体小鼠体重明显下降，表明免疫介导的致病性。7．克隆了汉滩病毒S和M基因组片段，分别编码核衣壳蛋白和包膜糖蛋白，并在哺乳动物细胞中表达。 8．编码聚合酶的整个L基因组片段被克隆。 L克隆的表达正在进行中。