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当前位置: 首页 > 文章 > REV Gp90亚单位疫苗联合核酸缓释佐剂对雏鸡的免疫保护作用 中国兽医学报 2017,37 (10) 1856-1861
Position: Home > Articles > Immunoprotection of recombinant Gp90 subunit vaccine combining with nucleic acid sustained releasing adjuvant against REV infection in chickens Chinese Journal of Veterinary Science 2017,37 (10) 1856-1861

REV Gp90亚单位疫苗联合核酸缓释佐剂对雏鸡的免疫保护作用

作  者：
袁霏;王康;郭道磊;马文杰;褚颖;郭慧君
单  位：
山东农业大学动物科技学院山东省生物工程与疫病防治重点实验室
关键词：
禽网状内皮组织增生症病毒;Gp90囊膜蛋白;CpG ODN;Poly(I:C);核酸缓释佐剂;免疫保护
摘  要：
主要探讨禽网状内皮组织增生症病毒(REV)亚单位疫苗联合CpG-ODN/Poly(I:C)缓释佐剂对雏鸡的免疫保护作用。根据已发表的REV囊膜蛋白Gp90的基因序列,设计引物,从感染REV的细胞中扩增出目的基因,构建原核重组表达载体,在Rosseta(E.coli)菌中诱导表达,对表达蛋白进行纯化和Western blot鉴定;制备的重组蛋白与CpG-ODN/Poly(I:C)缓释佐剂联合接种7日龄雏鸡,2次加强免疫后第2周进行REV攻毒,接种后每周检测血清抗体和攻毒后检测REV病毒血症,评价免疫保护效果。结果显示,成功构建重组原核表达质粒,并获得相对分子质量为51 000的重组表达产物,该产物能够与REV单克隆抗体特异性结合;与CpG-ODN/Poly(I:C)缓释佐剂联合接种后能够诱导雏鸡产生较单一免疫Gp90蛋白抗体效价显著增加的血清抗体,经二次加强免疫全部鸡只产生抗体;攻毒后接种雏鸡病毒血症为0%(0/13),而对照组和单一免疫Gp90蛋白组分别为69%(9/13)和15%(2/13)。结果表明,本研究制备的REV Gp90重组蛋白联合CpG-ODN/Poly(I:C)缓释佐剂对雏鸡具有完全的保护力,这为开展REV疫苗防控提供科学数据。
译  名：
Immunoprotection of recombinant Gp90 subunit vaccine combining with nucleic acid sustained releasing adjuvant against REV infection in chickens
作  者：
YUAN Fei;WANG Kang;GUO Dao-lei;MA Wen-jie;CHU Ying;GUO Hui-jun;College of Veterinary Medicine,Research Center for Animal Disease Control Engineering Shandong Province;Station of Veterinary,Animal Husbandry Bureau of Dongping County;
单  位：
College of Veterinary Medicine,Research Center for Animal Disease Control Engineering Shandong Province%Station of Veterinary,Animal Husbandry Bureau of Dongping County
关键词：
REV;;Gp90 protein;;CpG ODN;;Poly(I:C);;nucleic acid sustained releasing adjuvant;;immunoprotection
摘  要：
The aim of this study was to evaluate the immunoprotection of recombinant REV Gp90 subunit vaccine combining with CpG-ODN/Poly(I:C)nucleic acid sustained releasing adjuvant against REV infection in chickens.The gp90 gene of REV strain was amplified by PCR and the recombinant plasmid containing the target gene(gp90)was constructed and expressed in Rosseta(Escherichia coli),the expressed products of Gp90 were purified and then confirmed with Western blot.The purified recombinant proteins combining with CpG ODN/Poly(I:C)sustained releasing adjuvant were inoculated into the chickens,two booster inoculations were conducted at an interval of 3 weeks;at the 2 nd week of the last inoculation,all the chickens were challenged.REV-antibody in serum in the vaccinated chickens was detected weekly,the viremia in plasma of each challenged chicken was detected by indirect immunoinfluorescence assay(IFA)mediated with monoclonal antibody(MAb)of REV.The results showed that the recombinant plasmid containing gp90 gene was successfully constructed and expressed into the recombinant protein with the size of 51 000,which can recognize MAb of REV.Compared with control group and Gp90 group,the antibody titers and the positive ratio of REV antibodies in Gp90+CpG/Poly(I:C)group were significantly increased after first inoculation and further improved by booster inoculation.At the third week post the second inoculation,all the chickens were induced to produce serum REV-antibody with higher titers.After challenged,all the chickens in Gp90+CpG/Poly(I:C)group were negative to REVviremia,but in other two groups,some chickens were positive to REV-viremia.These data suggested the sustained releasing adjuvant containing CpG-ODN and Poly(I:C)can enhance the antibody responses to recombinant Gp90 protein antigen and the protection against REV infection.

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