【摘要】 西尼罗病毒(WNV)是一种可以导致人与动物死亡的人兽共患病的虫媒病毒。为构建表达WNV Pr M/E蛋白的重组新城疫病毒(NDV)La Sota疫苗株并对其免疫原性进行评价,本研究利用反向遗传学操作技术将人工合成的WNV pr M/E基因插入至NDV基因组P和M基因之间,构建重组病毒r La-WNV-Pr M/E。以重组病毒分别感染BHK-21细胞和免疫小鼠,利用RT-PCR、间接免疫荧光、ELISA和流式细胞术(FACS)检测Pr M/E蛋白的表达情况及免疫效果。结果显示,WNV-Pr M/E蛋白在BHK-21细胞中获得正确表达,免疫小鼠可以诱导其产生高水平的抗Pr M/E蛋白特异性Ig G,并且诱导产生WNV E蛋白表位特异性CD4+和CD8+T细胞免疫反应。以上结果表明本研究构建的r La-WNV-Pr M/E可以作为一种具有前瞻性和储备性且安全有效的西尼罗热防控候选疫苗,对我国应对该病的潜在威胁具有十分重要的意义。更多还原

【Abstract】 West Nile Fever is an entomophilous zoonosis caused by West Nile virus(WNV), which causes the death of humans and animals. This study generated a recombinant Newcastle disease virus(NDV) of La Sota vaccine strain expressing the codon-optimized envelope protein(E) and premembrane protein(Pr M) of WNV by reverse genetics, designated r La-WNV-Pr M/E,and evaluation of immune in mice which produced humoral and cellular immune responses was to test the immunogenicity of the recombinant virus. RT-PCR, indirect immunofluorescence(IFA), ELISA and Fluorescence Activated Cell Sorting(FACS) were used for measuring the expression of WNV-Pr M/E and immune response. Results showed that the r La-WNV- Pr M/E expressed the Pr M/E protein effectively and induced high levels of specific Ig G against Pr M/E and specific CD4+and CD8+T cells anti-WNV E protein epitope in immune response. These results demonstrated that r La-WNV-Pr M/E was efficient to stimulate high level humoral and cellular immunoresponses, which could be used a potential vaccine candidate against WNV infection.更多还原