期刊文献+

HIV-1 C亚型密码子优化gp120负载人DC疫苗的构建及体外功能

Construction and function research of dendritic cell vaccine loaded with Codon usage-optimized gp120 of HIV-1 subtype C in vitro
下载PDF
导出
摘要 目的构建HIV-1C亚型gp120负载人树突状细胞(dentriti ccell,DC)疫苗,并对其体外功能进行初步检测。方法利用Amaxa细胞核转染技术将pcDNA3.1-gp120质粒转染至人成熟DC,以Western blot检测gp120的表达。通过流式细胞仪检测DC表面共刺激分子的变化、混合淋巴细胞反应、CD8+T细胞表面活化分子CD25的表达及其分泌IFN-γ的变化。结果通过Western blot检测,gp120在DC中得到了正确表达。经流式细胞仪检测,DC表面分子CD80表达率由刺激前的33.34%上升至43.20%,CD86表达率由刺激前的60.08%上升至90.34%;负载gp120DC刺激淋巴细胞增殖率为86.72%;CD8+T细胞表面分子CD25表达率由刺激前的5.27%上升至74.21%,IFN-γ的表达率达37%。结论负载了HIV-1gp120的人树突状细胞能够显著刺激淋巴细胞的增殖、增强CD8+T细胞表面活化分子CD25表达以及促进CD8+T细胞分泌IFN-γ,为下一步DC治疗性疫苗的体内研究奠定基础。 This study is aimed to construct a therapeutic vaccine containing HIV-1 subtype C gp120 and test the function in vitro. Mature DCs were transfected by recombined expression plasmid pcDNA3.1-gp120. Then the expressions of gp120 were identified by Western blot; flow cytometry was applied to analyze the expression levels of costimulatory molecules CD80 and CD86,the mixed lymphocyte reaction,the CD25 production and IFN-γ secretion. We found that gp120 was correctly expressed in DCs; CD80 and CD86 increased from 33.34% to 43.20% and 60.08% to 90.34%,respectively; T cell proliferation rate was 86.72%. The expression rate of CD25 increased from 5.27% to 74.21%. IFN-γ secreted by CD8+ T cells reached a high level of 37%. We conclude that T cell proliferation is promoted by gp120-loaded DCs,and The CD25 production and IFN-γ secretion are significantly up-regulated,which provides a base for studying the function of the vaccine in vivo.
出处 《免疫学杂志》 CAS CSCD 北大核心 2010年第9期741-746,共6页 Immunological Journal
基金 国家自然科学基金(30471605) 中国科学院知识创新工程重要方向(KSCX1-YW-R-15,KSCX2-YW-R-092) 云南省科技基础条件平台计划(2006PT08)
关键词 HIV-1 C亚型 密码子优化 GP120 树突状细胞 HIV-1 Subtype C Codon usage-optimized gp120 Dendritic cell
  • 引文网络
  • 相关文献

参考文献27

二级参考文献70

共引文献21

;
使用帮助 返回顶部