期刊文献+
任意字段
题名或关键词
题名
关键词
文摘
作者
第一作者
机构
刊名
分类号
参考文献
作者简介
基金资助
栏目信息

表达猪白介素2基因重组腺病毒的构建及其免疫增强作用 被引量：3

Construction and Immunoenhancement of a Recombinant Adenoviruses Encoding Porcine Interleukin 2
在线阅读 下载PDF
导出
摘要 以猪白介素2基因(pIL-2)为研究对象构建表达pIL-2的重组腺病毒,并研究其作为细胞因子佐剂的免疫增强作用。用RT-PCR的方法扩增pIL-2基因,将pIL-2基因克隆到腺病毒穿梭载体AdTrack中,构建出重组腺病毒穿梭质粒AdTrack-pIL-2,然后在大肠杆菌BJ5183内和骨架载体AdEasy同源重组,获得重组腺病毒骨架载体AdEasy-pIL-2,转染HEK293细胞,包装出重组腺病毒Ad-pIL-2,并通过兔体交互试验来检测Ad-pIL-2的免疫增强作用。经PCR、RT-PCR和Western blot检测,成功构建了Ad-pIL-2,病毒滴度可以达到108.25pfu/mL,并通过兔体交互免疫试验证明Ad-pIL-2可以提高兔的抗体水平。用腺病毒构建的Ad-pIL-2在兔体上具有免疫增强作用,为下一步的猪体试验研究和细胞因子佐剂的开发奠定基础。 The recombinant adenovirus carrying porcine interleukin 2（pIL-2） was constructed and the immunologic enhancement of it was studied as a cytokine adjuvant.The pIL-2 gene fragment was amplified by RT-PCR and was inserted into adenovirus shuttle vector AdTrack-CMV to produce the AdTrack-pIL-2.Linearized by Pme I,it was transferred into E.coli BJ5183 cells where AdEasy-pIL-2 was generated.The recombinant adenovirus Ad-pIL-2 was propagated after transfecting AdEasy-pIL-2 into HEK293 cells,and the immunoenhancement of it was evaluated on rabbits then.pAd-pIL-2 could carry and express pIL-2 protein by identification of PCR,RT-PCR and Western-blot.The titer of it was 10^8.25 pfu/mL.It could improve the IgG Level of rabbits compared to the group without being inoculated.Recombinant adenovirus vector Ad-pIL-2 was constructed successfully and the immune enhancing effect of it was evaluated on rabbits,which may lay a foundation for the further study of the effect on pigs as a cytokine adjuvant.
作者 何雷 张彦明 向华 唐青海 徐彦召 杨小云 王静 代晨
机构地区 西北农林科技大学动物医学院
出处 《西北农业学报》 CAS CSCD 北大核心 2010年第11期1-7,共7页 Acta Agriculturae Boreali-occidentalis Sinica
基金 "863"重大项目"家畜重要病毒病基因工程疫苗研究和创制"(2006AA10A204)
关键词 猪白介素2 重组腺病毒 细胞因子 佐剂 Porcine interleukin 2 Recombinant adenovirus Cytokine Adjuvant
分类号 S852.5 [农业科学—基础兽医学]
  • 相关文献

参考文献15

  • 1Malek T R.The main function of IL-2 is to promote the development of T regulatory cells[J].Journal of Leukocyte Biology,2003,74(12):961-965.
  • 2Bukreyev A,Whitehead S S,Prussin C,et al.Effect of coexpression of interleukin-2 by recombinant respiratory syncytial virus on virus replication,immunogenicity,and production of other cytokines[J].Journal of Virology,2000,74 (15):7 151-7 157.
  • 3Hyun S,Lillehoj,Xi C D,et al.Resistance to Intestinal Coccidiosis following DNA immunization with the cloned 3-1E Eimeria gene plus IL-2,IL-15 and IFN-γ[J].Avian Dis,2005,49(1):112-117.
  • 4Kim J J,Simbiri K A,Sin J I,et al.Cytokine molecular adjuvants modulate immune responses induced by DNA vaccine constructs for HIV-1 and SIV[J].Interferon Cytokine Res,1999,19(1):77-84.
  • 5Zhang H Y,Sun S H,Guo Y J,et al.Optimization strategy for plasmid DNAs containing multiple-epitopes of foot-and-mouth disease virus by cis-expression with IL-2[J].Vaccine,2008,26(6):769-777.
  • 6Xin K Q,Hamajima K,Sasaki S,et al.Intranasal administration of human immunodeficiency virus type-1 (HIV-1) DNA vaccine with interleukin-2 expression plasmid enhances cell-mediated immunity against HIV-1[J].Immunology,1998,94(3):438-444.
  • 7Hammond J M,Jansen E S,Morrissy C J,et al.Protection of pigs against ' in contact' challenge with classical swine fever following oral or subcutaneous vaccination with a recombinant porcine adenovirus[J].Virus Res,2003,97 (2):151-157.
  • 8Toledo J R,Sanchez O,Montesino R,et al.Highly protective E2-CSFV vaccine candidate produced in the mammary gland of adenoviral transduced goats[J].Journal of Biotechnology,2008,133(3):370-376.
  • 9Kaufman R J.Overview of vector design for mammalian gene expression[J].Mol Biotechnol,2000,16(2):151-160.
  • 10Tatsis N,Ertl H C.Adenoviruses as vaccine vectors[J].Molecular therapy,2004,10 (4):616-629.

二级参考文献52

  • 1Kim JJ,Yang JS,Manson KH,et al. Modulation of antigen-specific cellular immune responses to DNA vaccine in rhesus macaques through the use of IL-2, IFN-gamma,or IL-4 gene adjuvant. Vaccine,2001,19(17-19) :2496-2505.
  • 2Gherardi MM, Ramirez JC, Esteban M. Towards a new generation of vaccines: the cytokine IL-12 as an adjuvant to enhance cellular immune responses to pathogens during prime-booster vaccination regimens. Histol Histopathol, 2001,16 (2): 655-667.
  • 3Okada E, Sasaki S, Ishii N , et al. Intranasal immunization of a DNA vacine with IL-12 and granulocyte-macrophage colony-stimulating factor (GM-CSF) expressing plasmids in liposomes induces strong mucosal and cell-mediated immune reponses against HIV-1 am
  • 4Dupre L,Kremer L,Wolowczuk I,et al. Immunostimulatory effect of IL-18-encoding plasmid in DNA vaccination against murine Schistosoma mansoni infection. Vaccine, 2001, 19 (11-12): 1373-1380.
  • 5Hanlon L, Argyle D, Bain D, et al. Feline leukemia virus DNA vaccine efficacy is enhanced by coadministration with interleukin-12(IL-12)and IL-18 expression vectors. J Virol,2001,75(18) :8424-8433.
  • 6Gurunthan S, Prussin C, Sacks DL, et al. Vaccine requirements for sustained cellular immunity to an intracellular parasitic infection.Nat Med, 1998,4 (12): 1409-1415.
  • 7Kim JJ, Ayyavoo V, Bagarazzi ML, et al. In vivo engineering of a cellular immune response by coadministration of IL-12 expressin vector with a DNA, immunogen. J Immunol, 1997, 158 (2): 816-826.
  • 8Lee YL, Tao MH, Chow YH, et al. Construction of vectors expressing bioactive heterodimeric and single-chain murine interleukin-12 for gene therapy. Hum Gene Ther, 1998,9 (4): 457-465.
  • 9Hoshino J,Naganurna F,Nagai R. Ventricular fibrillation triggered by a ruptured sinus of Valsalva aneurysm. Heart, 1998,80 (2):203-204.
  • 10Rakhmilevich AL, et al. Cytokine gene therapy of cancer using gene gun technology:superior antitumor activity of interleukin-12. Hum Gene Ther, 1997,8(11): 1303-1311.

共引文献10

同被引文献69

引证文献3

二级引证文献13

西北农业学报
2010年 第11期

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部