利用假病毒技术筛选H5N1亚型禽流感病毒中和活性单克隆抗体  

Development of neutralizing monoclonal antibodies against H5N1 avian influenza virus by using pseudotyped HIV/HA virus

作　　者：张卓[1,2] 刘明[2] 郑世民[1] 

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,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2015年第7期545-548,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金(30371066)

摘　　要：为筛选H5N1禽流感病毒(AIV)血凝素蛋白(HA)特异性中和抗体,本研究构建了表达HA蛋白的重组质粒,利用该重组质粒及缺失表达人免疫缺陷病病毒1型(HIV-1)囊膜蛋白基因的骨架质粒,构建了表面整合有H5亚型AIV HA蛋白的HIV假病毒。利用该假病毒系统,筛选得到一株具有中和活性的单克隆抗体(MAb)。经测定,该MAb与纯化的全病毒具有较好的反应性,其对HIV-H5HA假病毒的中和效价为64。中和试验表明该MAb能够有效阻断野生型病毒对鸡胚的感染。本研究结果为开发H5N1 AIV的被动免疫治疗方法奠定了基础,同时对亚单位疫苗的研制也具有一定的意义。In order to prepare neutralizing antibodies against H5N1 avian influenza virus（AIV）, we first constructed a recombinant plasmid expressing HA gene of H5N1 AIV, then pseudotyped virus was packaged with this recobinant plasmid and Env-deficient HIV-1 gemome. By using this pseudotyped virus system, we screened a monoclonal antibody（MAb） with neutralizing activity（named 5E7）. This MAb showed positive reactivity with purified whole virus protein. Pseudotyped virus-based microneutraliztion assay showed that the MAb had a neutralizing titer of 64. In addition, it was shown that the MAb was able to block virus infection in chicken embryo eggs. Results of the study provided the basis for immunotherapy against H5N1 AIV infection, and is also valuable for subunit vaccine design.

关 键 词：H5N1禽流感病毒 假病毒 中和抗体 

分 类 号：S852.65[农业科学—基础兽医学]