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Development of multily attenuated Sendai virus vaccine by means of reverse genetics

利用反向遗传学开发多重减毒仙台病毒疫苗

基本信息

批准号：
10680785
负责人：
ITOH Masae
金额：
$ 0.58万
依托单位：
Osaka Prefectural Institute of Public Health
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10680785/
关键词：
Sendai virus pathogenicity apoptosis necrosis interferon

项目摘要

(1) Identification of Sendai virus genes which determine mouse pathogenicity.Comparing an attenuated mutant virus (MVC11) with a highly pathogenic wild-type virus (M1), we showed the single point mutation from Phe to Ser at the 170th position of the C protein abolished the virulence of Sendai virus against mice. Involvement of the C protein in pathogenicity was confirmed by examining some recombinant viruses recovered by means of reverse genetics which were lacking one or two of the set of the C proteins (C', C, Y1 and Y2).(2) Mechanism of attention of the c protein mutant.1) Interferon sensitivity: When MVC11 was infected to interferon α/β receptor (IFNR)-knock-out mice (A129), mice exhibited only slight decrease of body weight. The result demonstrated that MVC11 was attenuated in A129 to the same degree as in ordinary mice possessing IFNR, suggesting that interferon sensitivity does not relate to attenuation of MVC11.2) Effect of death of infected cells: Attenuated viruses with mutations in the C protein like MVC11 induced necrosis as well as apoptosis. Death of the infected cells caused interruption of progeny virus production afterwards, and resulted in attenuation of viral pathogenicity. On the other hand, pathogenic viruses like M1 did not demonstrate significant cytopathic effect and released progeny virus continuously for a long time after infection. In the presence of caspase inhibitor which suppressed apoptosis but not necrosis, MVC11-infected cells died rapidly. These results suggested that necrosis plays an important role in causing death to attenuated virus-infected cells.(3) Growth of recombinant Sendai virus: Recombinant virus of Sendai virus obtained from cDNA of the attenuated laboratory strain (Z), the C gene of which was substituted by that of MVC11 did not grow efficiently. Based on this observation, we are trying to introduce mutations into the F and V genes of MVC11 with the aim to establish multiply attenuated virus.

(1)仙台病毒对小鼠致病性基因的鉴定：将一株弱毒突变病毒（MVC 11）与一株高致病性野生型病毒（M1）进行比较，发现仙台病毒C蛋白第170位由Phe突变为Ser的单点突变使其对小鼠的致病性丧失。通过检查一些重组病毒证实了C蛋白参与致病性，所述重组病毒通过反向遗传学回收，所述重组病毒缺少一种或两种C蛋白（C '，C，Y1和Y2）。(2)c蛋白突变体的注意机制。1）干扰素敏感性：当MVC 11感染干扰素α/β受体（IFNR）敲除小鼠（A129）时，小鼠仅表现出轻微的体重下降。结果表明，MVC 11在A129中的减毒程度与在具有IFNR的普通小鼠中的减毒程度相同，这表明干扰素敏感性与MVC 11的减毒无关。2）感染细胞死亡的影响：在C蛋白中具有突变的减毒病毒如MVC 11诱导坏死以及凋亡。感染细胞的死亡导致子代病毒生产中断，导致病毒致病性减弱。另一方面，致病性病毒如M1没有表现出明显的细胞病变效应，并且在感染后较长时间内持续释放子代病毒。在存在抑制凋亡但不抑制坏死的半胱天冬酶抑制剂的情况下，MVC 11感染的细胞迅速死亡。这些结果表明，坏死在致弱病毒感染细胞死亡中起重要作用。(3)重组仙台病毒的生长：从减毒实验室株（Z）的cDNA获得的仙台病毒的重组病毒，其C基因被MVC 11的C基因取代，不能有效生长。基于这一观察结果，我们试图将突变引入MVC 11的F和V基因，目的是建立多重减毒病毒。