表达西尼罗病毒囊膜糖蛋白PrM/E重组新城疫病毒的构建及其免疫原性研究  

Construction of a recombinant Newcastle disease virus expressing West Nile virus Pr M/E and evaluation of its immunogenicity in mice

作　　者：杨洁[1] 温志远[2] 葛金英[2] 王金良[2] 华荣虹[2] 邵钰[2] 步志高[2] 

机构地区：[1]东北农业大学动物医学学院,黑龙江哈尔滨150030 [2]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/农业部兽医公共卫生重点开放实验室,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2015年第8期585-589,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家科技支撑计划"严重威胁的外来动物疫病免疫防控储备技术"(2013BAD12B05)

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。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 LaSota vaccine strain expressing the codon-optimized envelope protein （E） and premembrane protein （PrM） of WNV by reverse genetics, designated rLa-WNV-PrM/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-PrM/E and immune response. Results showed that the rLa-WNV- PrMfE expressed the PrM/E protein effectively and induced high levels of specific IgG against PrM/E and specific CD4＋ and CD8＋ T cells anti-WNV E protein epitope in immune response. These results demonstrated that rLa-WNV-PrM/E was efficient to stimulate high level humoral and cellular immunoresponses, which could be used a potential vaccine candidate against WNV infection.

关 键 词：西尼罗病毒 PrM/E蛋白 重组新城疫病毒 

分 类 号：S852.65[农业科学—基础兽医学]