[1]李国新,郑宝亮,金媛媛,等.分子佐剂C3d增强猪流感病毒HA-DNA疫苗免疫原性的研究[J].中国预防兽医学报,2008,(10):814-819.
 LI Guo-xin,ZHENG Bao-liang,JIN Yuan-yuan,et al.C3d-induced immune enhancement of the swine influenza HA expressed by a DNA vaccine[J].Chinese journal of preventive veterinary medicine,2008,(10):814-819.
点击复制

分子佐剂C3d增强猪流感病毒HA-DNA疫苗免疫原性的研究
分享到:

《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2008年10期
页码:
814-819
栏目:
免疫学
出版日期:
2008-10-01

文章信息/Info

Title:
C3d-induced immune enhancement of the swine influenza HA expressed by a DNA vaccine
文章编号:
1008-0589(2008)10-0814-06
作者:
李国新12郑宝亮3金媛媛2田志军2周艳君2童光志4*
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/猪传染病研究室,黑龙江 哈尔滨 150001;
3. 郑州牧业工程高等专科学校 生物工程系,河南 郑州 450011;
4. 中国农业科学院上海兽医研究所,上海 200232
Author(s):
LI Guo-xin12 ZHENG Bao-liang3 JIN Yuan-yuan2 TIAN Zhi-jun2 ZHOU Yan-jun2 TONG Guang-zhi4*
1. College of Animal Medicine, Northeast Agricultural University, Harbin 150030, China;
2. Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China;
3. Zhengzhou College of Animal Husbandry Engineering, Zhengzhou 450011, China;
4. Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200232, China
关键词:
猪流感病毒DNA疫苗分子佐剂C3d
Keywords:
swine influenza virus DNA vaccine molecular adjuvant C3d
分类号:
S852.659;S858.28
文献标志码:
A
摘要:
为了探讨分子佐剂C3d对猪流感病毒HA-DNA疫苗免疫原性的影响,本研究构建了3种表达不同形式HA的重组质粒,分别为:表达分泌型HA的质粒p-sHA、表达全长HA的质粒p-tmHA和融合表达3拷贝鼠C3d与HA的质粒p-sHA/mC3d3。3种质粒与空载体pCAGGS分别免疫BALB/c小鼠后,采用ELISA和HI试验检测抗体水平。结果显示,免疫后第8周,p-sHA组与p-tmHA组的抗体水平没有明显差别,而p-sHA/mC3d3免疫组的抗体水平显著高于这两组,说明分子佐剂C3d增强了抗体应答水平。淋巴细胞增殖试验及细胞因子检测实验结果显示,免疫后第8周,p-tmHA诱导了更强的淋巴细胞增殖反应和更高水平的IFN- ,而p-sHA/mC3d3诱导了更高水平的IL-4,说明HA表达形式的变化影响了免疫反应的类型,C3d与HA融合后通过诱导IL-4的产生而使免疫反应倾向于Th2型。总之,本研究证实3拷贝分子佐剂C3d与分泌型HA融合后显著提高了DNA疫苗诱导的抗体水平。这提示质粒p-sHA/mC3d3免疫可能对接种动物提供高水平的保护,有希望成为抗猪流感病毒的候选疫苗。
Abstract:
To evaluate the effects of C3d on the immunogenicity of the swine influenza HA expressed by a DNA vaccine. Plasmid encoding secretive HA (p-sHA), complete HA (p-tmHA) or secretive HA-C3d fusion prtein (p-sHA/mC3d3) were constructed from H3N2 subtype of swine influenza virus and administered to mice, respectively. The immune response was monitored by ELISA and hemagglutination inhibition (HI) assays. p-sHA/mC3d3 was shown to induce higher level of anti-HA antibodies compared to p-sHA or p-tmHA. Furthermore, C3d fusion increased Th2-biased immune response by inducing IL-4 production. The IL-4 level in splenocytes from mice received p-sHA/mC3d3 was three times higher than those vaccinated with p-sHA or p-tmHA. These results showed that the murine C3d gene fused to the truncated HA gene could significantly enhance antibody responses to hemagglutinin, and the plasmid DNA expressing sHA/mC3d3 fusion proteins may represent an effective candidate vaccine against swine influenza virus.

参考文献/References:

[1] Brown I H. The pig as an intermediate host for influenza A vir- uses between birds and humans[J]. International Congress Series, 2001, 1219: 173-178.
[2] Macklin M D, McCabe D, McGregor M W, et al. Immunization of pigs with a particle-mediated DNA vaccine to influenza A virus protects against challenge with homologous virus [J]. J Virol, 1998, 72(2): 1491-1496.
[3] Larsen D L, Karasin A, Olsen C W. Immunization of pigs against influenza virus infection by DNA vaccine priming followed by killed-virus vaccine boosting [J]. Vaccine, 2001, 19(20-22): 2842-2853.
[4] Carroll M C. The role of complement in B cell activation and tolerance [J]. Adv Immunol, 2000, 74: 61-88.
[5] Dempsey P W, Allison M E, Akkaraju S, et al. C3d of compl- ement as a molecular adjuvant: bridging innate and acquired immunity [J]. Science, 1996, 271: 348-350.
[6] Ross T M, Xu Y, Bright R A, et al. C3d enhancement of anti- bodies to hemagglutinin accelerates protection against influenza virus challenge [J]. Nat Immunol, 2000, 1(2): 127-131.
[7] Green T D, Newton B R, Rota P A, et al. C3d enhancement of neutralizing antibodies to measles hemagglutinin [J]. Vaccine, 2001, 20: 242-248.
[8] Mitchell J A, Green T D, Bright R A, et al. Induction of hetero- subtypic immunity to influenza A virus using a DNA vaccine expressing hemagglutinin-C3d fusion proteins [J]. Vaccine, 2003, 21(9-10): 902-914.
[9] Liu F, Mboudjeka I, Shen S, et al. Independent but not synergistic enhancement to the immunogenicity of DNA vaccine expressing HIV-1 gp120 glycoprotein by codon optimization and C3d fusion in a mouse model [J]. Vaccine, 2004, 22(13-14): 1764-1772.
[10] Tong T Z, Fan HY, Tan Y D, et al. C3d enhanced DNA vacci- nation induced humoral immune response to glycoprotein C of pseudorabies [J]. Biochem Biophys Res Commun, 2006, 347(4):845-851.
[11] Fan H, Tong T, Chen H, Guo A. Immunization of DNA vaccine encoding C3d-VP1 fusion enhanced protective immune response against foot-and-mouth disease virus [J]. Virus Genes, 2007, 35(2): 347-357.
[12] Li G X, Qiu H J, Han C G, et al. Vaccination of plasmid DNA encoding somatostatin gene fused with GP5 gene of porcine reproductive and respiratory syndrome virus induces anti-GP5 antibodies and promotes growth performance in immunized pigs [J]. Agricultural Sciences in China, 2006, 5(3): 234-240.
[13] 陈艳,辛晓光,杨焕良,等. 猪流感抗体间接ELISA检测方法的建立[J]. 中国预防兽医学报,2007,29(4):308-311.
[14] Tian Z J, Zhou G H, Zheng B L, et al. A recombinant pseudo- rabies virus encoding the HA gene from H3N2 subtype swine influenza virus protects mice from virulent challenge [J]. Veteri- nary Immunol Immunopathol, 2006, 111: 211-218.
[15] Bounous D I, Campagnoli R P, Brown J. Comparison of MTT colorimetric assay and tritiated thymidine uptake for lymphocyte proliferation assays using chicken splenocytes [J]. Avian Dis, 1992, 36(4): 1022-1027.
[16] Wesley R D, Tang M, Lager K M. Protection of weaned pigs by vaccination with human adenovirus 5 recombinant viruses expre- ssing the hemagglutinin and the nucleoprotein of H3N2 swine influenza virus [J]. Vaccine, 2004, 22(25-26): 3427-3434.
[17] Endo A, Itamura S, Iinuma H, et al. Homotypic and heterotypic protection against influenza virus infection in mice by recombi- nant vaccinia virus expressing the haemagglutinin or nucleoprotein of influenza virus [J]. J Gen Virol, 1991, 72(Pt3): 699-703.
[18] Tang M, Harp J A, Wesley R D. Recombinant adenovirus enco- ding the HA gene from swine H3N2 influenza virus partially protects mice from challenge with heterologous virus: A/HK/1/68 (H3N2) [J]. Arch Virol, 2002, 147(11): 2125-2141.
[19] Haas K M, Toapanta F R, Oliver J A, et al. Cutting edge: C3d functions as a molecular adjuvant in the absence of CD21/35 expression [J]. J Immunol, 2004, 172(10): 5833-5837.
[20] Suradhat S, Braun R P, Lewis P J, et al. Fusion of C3d molecule with bovine rotavirus VP7 or bovine herpesvirus type 1 glycopro- tein D inhibits immune responses following DNA immunization [J]. Vet Immunol Immunopathol, 2001, 83(1-2): 79-92.
[21] Gor D O, Ding X, Li Q, et al. Genetic fusion of three tandem copies of murine C3d sequences to diphtheria toxin fragment B elicits a decreased fragment B-specific antibody response[J]. Immunol Lett, 2006, 102(1): 38-49.
[22] Mongini P K, Vilensky M A, Highet P F, et al. The affinity threshold for human B cell activation via the antigen receptor complex is reduced upon co-ligation of the antigen receptor with CD21 (CR2) [J]. J Immunol, 1997, 159(8): 3782-3791.
[23] Ross T M, Xu Y, Green T D, et al. Enhanced avidity maturation of antibody to human immunodeficiency virus envelope: DNA vaccination with gp120-C3d fusion proteins [J]. AIDS Research and Human Retroviruses, 2001, 17(9): 829-835.

相似文献/References:

[1]王国俊,熊 杰,李俊平,等.H5亚型禽流感病毒A/Duck/Anhui/1/2006 (Clade 2.3)密码子优化HA基因DNA疫苗的免疫保护效力研究[J].中国预防兽医学报,2010,(02):116.
 WANG Guo-jun,XIONG Jie,LI Jun-ping,et al.Protective efficacy of DNA vaccine encoding codon-optimized HA gene of H5 subtype avian influenza virus A/Duck/Anhui/1/2006(Clade 2.3)[J].Chinese journal of preventive veterinary medicine,2010,(10):116.
[2]郭利敏,乔传玲*,陈 艳,等.利用真核表达的猪流感病毒NP蛋白制备其特异性的单克隆抗体[J].中国预防兽医学报,2010,(06):460.
 GUO Li-min,QIAO Chuan-ling*,CHEN Yan,et al.Preparation of the monoclonal antibodies using swine influenza virus NP protein expressed by eukaryotic expression vector[J].Chinese journal of preventive veterinary medicine,2010,(10):460.
[3]石星明,王 玫,张 晶,等.CpG基序对新城疫病毒 DNA免疫效果的影响[J].中国预防兽医学报,2010,(07):554.
 SHI Xing-ming,WANG Mei,ZHANG Jing,et al.Effects of CpG oligonucleotides on immune enhancement on NDV DNA immunization[J].Chinese journal of preventive veterinary medicine,2010,(10):554.
[4]罗 俊,刘业兵*,陈 坚,等.双重RT-PCR方法检测猪流感病毒和猪繁殖与呼吸综合征病毒[J].中国预防兽医学报,2010,(09):687.
 LUO Jun,LIU Ye-bing*,CHEN Jian,et al.Establishment of a duplex RT-PCR for the detection of swine influenza virus and porcine reproductive and respiratory syndrome virus[J].Chinese journal of preventive veterinary medicine,2010,(10):687.
[5]张鹏超,师小潇,哈 卓,等.猪流感病毒SYBR GreenⅠ定量RT-PCR检测方法的建立及应用[J].中国预防兽医学报,2010,(10):768.[doi:10.3969/j.issn.1008-0589.2010.10.06]
 ZHANG Peng-chao,SHI Xiao-xiao,HA Zhuo,et al.Establishment of SYBR GreenⅠreal time RT-PCR for detecting swine influenza virus[J].Chinese journal of preventive veterinary medicine,2010,(10):768.[doi:10.3969/j.issn.1008-0589.2010.10.06]
[6]侯贤宏,魏永伟,章晓栋,等.IBDV VP2/4/3与ChIL-18融合基因真核表达质粒和基因共表达质粒的免疫原性比较[J].中国预防兽医学报,2010,(10):785.[doi:10.3969/j.issn.1008-0589.2010.10.10]
 HOU Xian-hong,WEI Yong-wei,ZHANG Xiao-dong,et al.The immunogenicity of co-expressing plasmid and fusion genes plasmid containing IBDV VP2/4/3 and ChIL-18 genes[J].Chinese journal of preventive veterinary medicine,2010,(10):785.[doi:10.3969/j.issn.1008-0589.2010.10.10]
[7]韩庆功,乔传玲*,杨焕良,等.猪源H9N2亚型流感病毒HA基因的序列测定及真核表达[J].中国预防兽医学报,2007,(12):916.
 HAN Qing-gong,QIAO Chuan-ling*,YANG Huan-liang,et al.Sequencing and eukaryotic expression of HA gene of H9N2 subtype influenza virus from swine[J].Chinese journal of preventive veterinary medicine,2007,(10):916.
[8]陈 艳,辛晓光,杨焕良,等.猪流感抗体间接ELISA检测方法的建立[J].中国预防兽医学报,2007,(04):308.
 CHEN Yan,XIN Xiao-guang,YANG Huan-liang,et al.Establishment of an indirect ELISA for detection of antibodies against swine influenza virus[J].Chinese journal of preventive veterinary medicine,2007,(10):308.
[9]郑宝亮,田志军,李 若,等.表达H3N2亚型猪流感病毒HA的重组伪狂犬病病毒对仔猪的免疫效力评价[J].中国预防兽医学报,2007,(02):115.
 ZHENG Bao-liang,TIAN Zhi-jun,LI Ruo,et al.Protection of piglets from virulent challenge after immunization with a recombinant pseudorabies virus expressing the HA gene of a H3N2 subtype swine influenza virus[J].Chinese journal of preventive veterinary medicine,2007,(10):115.
[10]宋勤叶,温立斌,杨汉春*,等.猪GM-CSF基因对猪圆环病毒2型DNA疫苗诱导仔猪免疫反应的调节作用[J].中国预防兽医学报,2007,(02):120.
 SONG Qin-ye,WEN Li-bin,YANG Han-chun*,et al.Modulation effects of porcine GM-CSF on immune responses in piglets induced by porcine circovirus type 2 DNA vaccine[J].Chinese journal of preventive veterinary medicine,2007,(10):120.

备注/Memo

备注/Memo:
收稿日期:2007-12-03
基金项目:国家863计划(2006AA10A204)和国家973计划(2005CB523200)
作者简介:李国新(1973~),男,黑龙江兰西人,博士研究生,从事动物病毒分子生物学研究.
*通讯作者:E-mail:gztong@hvri.ac.cn
更新日期/Last Update: 2010-11-24