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Development of new serodiagnostic method for hemorrhagic fever with renal syndrome(HFRS) and vaccine development against HFRS.

肾综合征出血热（HFRS）血清学诊断新方法的开发及HFRS疫苗的开发。

基本信息

批准号：
59880005
负责人：
YAMANISHI Koichi
金额：
$ 6.14万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Developmental Scientific Research
财政年份：
1984
资助国家：
日本
起止时间：
1984 至 1986
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-59880005/
关键词：
Hemorrhagic fever with renal syndrome HI ELISA NT HFRS virus Monoclonal antibody ワクチン HI法マクロファージ

项目摘要

We have been engaged for the last three years in the development of the method for the serodiagnosis of hemorrhagic fever with renal syndrome(HFRS) virus, and also on the preliminary study of inactivated vaccine against HFRS.1)development of new diagnostic method for HFRS; The immunofluorescent (IF) technique has been used to detect antibody against HFRS virus in human and rodent sera. We started to develop new serodiagnostic methods such as neutralizing antibody (NT) test, hemagglutination inhibition (HI) test, and ELISA test. First, peroxidase-antiperoxidase staining was applied for counting foci induced by HFRS virus. This method is more convenient than plaque formation method for NT test. Second, hemagglutinating antigen was prepared from HFRS virus-infected cell fluid, and this antigen was applied for HI test. Strong cross-reactions among various virus strains were demonstrated by HI test. Finally, antibody captured ELISA test was developed, and IF, HI and ELISA tests were compared to detect antibodies. ELISA and IF tests were more sensitive than HI test, but HI test was easier than other two.2)development of vaccine: we have isolated monoclonal antibodies to analyse antigens of HFRS viruses. Two glycoproteins(G1, G2) were indentified on the viral virion, which correlated with NT and HI activities. Next, various HFRS viruses were inoculated to newborn mice or rats, and it could be found that the virulence of HFRS viruses is variable, depending on the species of infected animals as well as the viruses. Finally, inactivated vaccine was prepared from mice brains, and NT antibodies were induced in mice immunized with vaccine. It can be expected to be used as a vaccine for human and rodent after further experiments in animals.

本实验室从事肾综合征出血热（HFRS）病毒血清学诊断方法的建立和灭活疫苗的初步研究。1）HFRS诊断新方法的建立：应用免疫荧光技术检测人和鼠血清中HFRS病毒抗体。我们开始开发新的血清学诊断方法，如中和抗体（NT）试验、血凝抑制（HI）试验和ELISA试验。首先，应用过氧化物酶-抗过氧化物酶染色法对HFRS病毒感染的病灶进行计数。本法比空斑形成法更简便。其次，从HFRS病毒感染细胞液中制备血凝抗原，并应用于HI试验。HI试验表明，各毒株间有较强的交叉反应。最后，建立了抗体捕获ELISA检测方法，并对IF、HI和ELISA检测抗体进行了比较。ELISA和IF试验较HI试验敏感，但HI试验较其他两种试验简便。2）疫苗的研制：我们分离了HFRS病毒抗原的单克隆抗体。病毒颗粒上有两种糖蛋白（G1、G2），与NT和HI活性相关。接着，将各种HFRS病毒接种给新生小鼠或大鼠，可以发现HFRS病毒的毒力是可变的，这取决于感染动物的种类以及病毒。最后，从小鼠脑中制备灭活疫苗，并在疫苗免疫的小鼠中诱导NT抗体。经进一步动物实验，可望作为人和啮齿类动物的疫苗。