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Analysis of virus-receptor interaction and its application in veterinary science

病毒-受体相互作用分析及其在兽医学中的应用

基本信息

批准号：
11460148
负责人：
TAGUCHI Fumihiro
金额：
$ 10.11万
依托单位：
National Institute of Neutoscience, NCNP
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2001
项目状态：
已结题

项目摘要

We have investigated the interaction of mouse hepatitis virus (MHV) and its receptor CEACAM1 (MHVR) by using soluble form of MHVR (soMHVR). soMHVR1 derived from MHV-susceptible BALB/c mouse showed high affinity and neutralizing activity to MHV, while soMHVR2 isolated from MHV-resistant SJL mouse was low in affinity and neutralization activity. We have isolated mutant virus resistant to neutralization (srr7) by soMHVR1 from MHV-JHMV. Srr7 infected as efficiently as wild type JHMV to cells expressing MHVR1, but did more than 2 logs less efficiently than wt virus to cells expressing MHVR2.This inefficient infection of srr7 was revealed to be due to its low fusogenicity, namely low capability to enter into these cells. These results suggested that the binding ofMHVR2 to wild type S protein triggered its conformational change, while it failed to do so for srr7 S proteins. We have also found that soMHVR1 activated or potentiated MHV infection to MHVR-deficient cells. This implied that binding of soMHVR converted fusion-negative S protein to fusion-positive phenotype. Using this system, we can further investigate in cell-free system the interaction of MHV and receptor as well as dynamics MHV particles following its interaction with receptor. Env protein ofmurine leukemia virus Fv-4r plays an important role for the resistance of mice to various leukemia viruses. We have conferred the resistance against leukemia virus to otherwise susceptible mice by expressing Env using retrovirus vectors. We have developed a highly pathogenic influenza A virus by passaging an avirulent virus through chickens. This conversion was accompanied with the mutations in HA protein, which resulted in enhanced cleavability of the protein. This suggested that HA cleavability influenced the pathogenicity of influenza virus.

本研究利用可溶性小鼠肝炎病毒（MHV）受体CEACAM 1（MHVR）研究了MHV与其受体的相互作用。来自MHV易感BALB/c小鼠的soMHVR 1对MHV具有高亲和力和中和活性，而来自MHV抗性SJL小鼠的soMHVR 2的亲和力和中和活性较低。我们已经从MHV-JHMV中分离出对soMHVR 1中和（srr 7）有抗性的突变病毒。Srr 7对表达MHVR 1的细胞的感染效率与野生型JHMV一样高，但对表达MHVR 2的细胞的感染效率比野生型病毒低2个对数以上。这些结果表明，MHVR 2与野生型S蛋白的结合引发了其构象变化，而与srr 7 S蛋白的结合则没有引发构象变化。我们还发现，soMHVR 1激活或增强MHV感染MHVR缺陷细胞。这意味着与soMHVR的结合将融合阴性S蛋白转化为融合阳性表型。利用该系统，我们可以在无细胞体系中进一步研究MHV与受体的相互作用以及MHV与受体相互作用后的动力学过程。小鼠白血病病毒Fv-4 r的Env蛋白在小鼠对多种白血病病毒的抵抗中起重要作用。我们已经通过使用逆转录病毒载体表达Env，赋予了对白血病病毒的抗性，否则易感小鼠。我们已经通过鸡传代无毒病毒开发了一种高致病性甲型流感病毒。这种转化伴随着HA蛋白中的突变，这导致蛋白质的可切割性增强。这表明HA的裂解能力影响流感病毒的致病性。