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| 传染性法氏囊病病毒VP2与C3d串联基因重组表达质粒的构建及其免疫功能研究 | |
| 引用本文： | 马金荣,欧阳伟,王永山,吴晓悠,朱向东,张海彬,范红结,王忠灿,唐雨德.传染性法氏囊病病毒VP2与C3d串联基因重组表达质粒的构建及其免疫功能研究[J].中国预防兽医学报,2012,34(6):471-475. |
| 作者姓名： | 马金荣 欧阳伟 王永山 吴晓悠 朱向东 张海彬 范红结 王忠灿 唐雨德 |
| 作者单位： | 1. 江苏省农业科学院兽医研究所农业部兽用生物制品工程技术重点实验室/国家兽用生物制品工程技术研究中心,江苏南京210014;南京农业大学动物医学院,江苏南京210095 2. 江苏省农业科学院兽医研究所农业部兽用生物制品工程技术重点实验室/国家兽用生物制品工程技术研究中心,江苏南京,210014 3. 南京农业大学动物医学院,江苏南京,210095 4. 南京军区军事医学研究所南京军区疾病预防控制中心,江苏南京,210002 |
| 基金项目： | 江苏省自然科学基金资助项目 |
| 摘 要： | 为提高传染性法氏囊病病毒(IBDV)VP2基因核酸疫苗的免疫效力,本研究根据已发表的鸡源补体C3d序列,设计并合成在5’端添加编码连接肽(Gly4Ser)2序列的C3d基因。用同尾酶BglⅡ和BamHⅠ构建含有3拷贝C3d与VP2基因融合的重组表达质粒pcDNA-VP2-3C3d。用脂质体法转染BHK21细胞,48 h后,westernblot分析表明,表达的重组蛋白为162 ku;间接免疫荧光试验检测转染细胞中具有特异性荧光。用pcDNA-VP2-3C3d与前期构建的pcDNA-VP2分别免疫2周龄SPF鸡,二免14 d后,间接ELISA法检测IBDV抗体效价,pcDNA-VP2-3C3d组抗体水平显著高于pcDNA-VP2组;MTT法检测鸡脾淋巴细胞增殖活性,pcDNA-VP2-3C3d组免疫诱导的特异性淋巴细胞增殖活性显著高于pcDNA-VP2组(p<0.05)。本研究表明C3d可以增强VP2基因免疫诱导的IBDV特异性体液和细胞免疫应答。 |
| 关 键 词： | 传染性法氏囊病病毒 VP2基因 鸡补体C3d 分子免疫佐剂 基因免疫 |
Construction and immune responses of the tandemed gene of infectious bursal disease virus VP2 and chicken complement C3d |
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| MA Jin-rong,OUYANG Wei,WANG Yong-shan,WU Xiao-you,ZHU Xiang-dong,ZHANG Hai-bin,FAN Hong-jie,WANG Zhong-can,TANG Yu-de.Construction and immune responses of the tandemed gene of infectious bursal disease virus VP2 and chicken complement C3d[J].Chinese Journal of Preventive Veterinary Medicine,2012,34(6):471-475. | |
| Authors: | MA Jin-rong OUYANG Wei WANG Yong-shan WU Xiao-you ZHU Xiang-dong ZHANG Hai-bin FAN Hong-jie WANG Zhong-can TANG Yu-de |
| Institution: | 1.National Center for Engineering Research of Veterinary Bio-products,Institute of Veterinary Medicine,Key Laboratory of Veterinary Biological Engineering and Technology,Ministry of Agricultural,Jiangsu Academy of Agricutural Sciences,Nanjing 210014,China; 2.College of Veterinary Medicine,Nanjing Agricultural University,Nanjing 210095,China; 3.Center for Disease Control and Prevention of Nanjing Command,Military Medical Institute of Nanjing Command,Nanjing 210002,China) |
| Abstract: | To enhance the immune efficacy of the DNA vaccine against the challenge of infectious bursal disease virus(IBDV),a three tandemed copies of chicken complement C3d gene(3C3d) with the linker sequence of(Gly4Ser)2 at 5’ end was synthesized and inserting into pUC57 vector.pcDNA-VP2-3C3d was constructed by insertion of the 3C3d tandem gene into pcDNA-VP2 and transfected into BHK21 cells for expression.Western blot result showed that the expressed fusion protein of VP2-3C3d was about 162 ku and reacted positively with IBDV polyclonal antibodies.Expression of VP2-3C3d was also confirmed by the indirect immunofluorescence assay.In addition,pcDNA-VP2-3C3d and pcDNA-VP2 were used to immunize 2-week-old SPF chicken.The antibody titrations and lymphocyte proliferative responses indicated that immunized with pcDNA-VP2-3C3d induced higher titers of IBDV antibodies and lymphocyte proliferation than that of immunized with pcDNA-VP2(p<0.05),indicating that C3d enhanced both the humoral and cellular immune response against IBDV. |
| Keywords: | infectious bursal disease virus VP2 gene chicken complement C3d molecular adjuvant gene vaccination |
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