表达H5亚型AIV血凝素的重组NDV构建及免疫原性  

Construction and Immunological Activity of Recombinant Newcastle Disease Virus Expressing H5 Hemagglutinin Gene of Avian Influenza Virus

作　　者：杨大光[1] 柴丽娜[1] 王新卫[1] 

机构地区：[1]河南农业大学牧医工程学院,郑州450002

出　　处：《现代生物医学进展》2007年第9期1429-1431,1423,共4页Progress in Modern Biomedicine

摘　　要：利用反向遗传技术获得表达H5亚型禽流感病毒(AIV)血凝素(HA)的新城疫病毒(NDV)。克隆NDV clone 30的全长基因,通过在NDV的融合蛋白基因和血凝素-神经氨酸酶(HN)基因之间插入编码高致病性AIV分离株A/chicken/italy/8/98(H5N2)的血凝素基因开放阅读框从而获得两株重组新城疫病毒NDVH5和NDVH5m。NDVH5感染的细胞可以检测到两种HA转录产物。对于重组病毒NDVH5m,NDV位于HA ORF的转录终止信号序列被沉默突变消除,产生2.7个全长HA转录产物的折叠,从而使修饰过的HA得到稳定地高表达。1日龄小鸡的脑内接种证实了两种重组病毒均无致病性。鸡群在NDVH5m诱导产生的NDV和H5亚型AIV HA特异性抗体的免疫力下能够免于致死剂量的NDV与高致病性AIV的感染。血清学研究结果表明NDVH5m免疫鸡群产生的抗体可结合NP蛋白抗体的检测从而用于区分免疫和感染AIV的动物。因此,NDVH5m重组病毒可作为抗NDV和AIV的"二联疫苗",也可成为控制AJ的标记疫苗。Newcastle disease virus （NDV）-expressing avian influenza virus （AIV） hemagglutinin （HA） of subtype H5 was constructed by reverse genetics. A cloned hill-length copy of the genome of the lentogenic NDV strain Clone 30 was used for insertion of the ORF encoding the HA of the highly pathogenic AIV isolate A/chicken/Italy/8/98 （HSN2） in the intergenic region between the NDV fusion and hemagglutinin-neuraminidase （HN） genes. Remarkably, two species of HA transcripts were detected in cells infected with the resultant NDVH5. In a second recombinant （NDVH5m）, a NDV transcription termination signal-like sequence located within the HA ORF was eliminated by silent mutations. Consequently, NDVHSm produced 2.7-fold more full-length HA transcripts, expressed higher levels of HA, and also incorporated more HA protein into its envelope than NDVHS. NDVHSm stably expressed the modified HA gene for 10 egg passages and both recombinants were found innocuous after intracerebral inoculation of 1-day-old chickens. Immunization of chickens with NDVHSm induced NDV and AIVHS-specific antibodies and protected chickens against clinical disease after challenge with a lethal dose of velogenic NDV or highly pathogenic AIV, respectively. Remarkably, shedding of influenza virus was not observed. Furthermore, immunization with NDVHSrn permitted serological discrimination of vaccinated and AIV field virus-infected animals based on antibodies against the nucleoprotein of AIV. Therefore, recombinant NDVH5m is suitable as a bivalent vaccine against NDV and AIV and may be used as marker vaccine for the control of avian influenza.

关 键 词：重组新城疫病毒 疫苗 禽流感 

分 类 号：S852.659[农业科学—基础兽医学]