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国家自然科学基金(30460050)

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发文基金:国家自然科学基金教育部“新世纪优秀人才支持计划”海南省教育厅科研基金更多>>
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小鼠VCAM-1真核表达质粒的构建及表达被引量:1
2007年
目的:构建小鼠血管细胞粘附分子-1(VCAM-1)基因真核表达质粒,了解该质粒是否能在真核细胞中有效表达。方法:以小鼠胚肝组织总RNA为模板,通过RT-PCR扩增小鼠VCAM-1全段基因cDNA,应用基因工程技术将扩增的cDNA片段插入pcDNA3.1(+)真核表达质粒,构建重组真核表达质粒pcDNA-mVCAM,经限制性内切酶酶切分析及测序鉴定正确后,用脂质体转染技术转染CHO细胞,用Western Blot鉴定该质粒是否能够在真核细胞中表达相应的目的蛋白质。结果:成功扩增了小鼠VCAM-1全长基因cDNA,酶切和测序证明正确构建了重组真核表达质粒pcDNA-mVCAM,West-ern Blot方法证实该质粒能在CHO真核细胞中正确表达目的蛋白。结论:成功构建了小鼠VCAM-1基因的真核表达质粒载体,为进一步研究VCAM-1的新功能和免疫治疗奠定了基础。
谭光宏宿静梅黄风迎王华黄用豪林映莹
关键词:血管细胞粘附分子-1真核表达质粒基因工程
Active immunotherapy of allergic asthma with a recombinant human interleukin-5 protein as vaccine in a murine model被引量:3
2007年
Background Eosinophils are highly related to allergic asthma inflammation. Interleukin (IL)-5 is the major chemokine of eosinophils, inhibition of the activity of IL-5 thus seems to be a potential approach to asthma therapy. The current study was performed to determine whether a recombinant human IL-5 protein as a xenogeneic vaccine has the capability of inducing anti-asthma activities. Methods Recombinant human IL-5 was used as a protein vaccine. Mouse asthma model was established to observe the anti-asthma activities. Lung histology was observed; eosinophils in blood and bronchoalveolar lavage were stained and counted, Airway hyperresponsiveness was determined by whole body plethysmograph. Antibody characters and cytokines were detected with enzyme linked immunosorbent assay (ELISA) and Western blot assay. Results Vaccination with recombinant human IL-5 protein as vaccine significantly reduced airway inflammation and airway hyperresponsiveness, and shifted the cytokine production from Th2 (IL-4) to Thl (INF-γ) in mice allergic-asthma model. Immunization with recombinant human IL-5 protein vaccine bypassed the immunological tolerance and induced production of polyclonal antibodies that were cross-reactive with murine IL-5. Conclusions Active immunization with xenogeneic homologous IL-5 may be a possible therapeutic approach to the treatment of asthma and potentially of other eosinophilic disorders.
TAN Guang—hongWANG Cai-chunHUANG Feng-yingWANG HuaHUANG Yong—haoLIN Ying-ying
关键词:ASTHMAINTERLEUKIN-5EOSINOPHIL
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