摘要
目的探讨HBV S-ecdCD40L融合基因修饰对树突状细胞(DC)功能的影响。方法分离健康成人外周血单个核细胞,GM-CSF、IL-4诱导培养树突状细胞,培养第5天,以脂质体介导转染,分为pcDNA3.1-S-ecdCD40L转染组、pcDNA3.1-S转染组、pcDNA3.1转染组及PBS对照组。转染48h收集DC及上清,流式细胞仪检测DC表面CD80、CD86、HLA-DR表达水平;CCK-8法检测混合淋巴细胞反应(MLR)中DC刺激同种异体淋巴细胞增殖的能力;酶联免疫吸附试验(ELISA)检测IL-12的分泌水平。结果与对照组比较,pcDNA3.1-S-ecdCD40L转染组DC表达CD80、CD86、HLA-DR水平增加,刺激同种异体淋巴细胞增殖的能力增强,分泌IL-12水平增高。结论 HBV S-ecdCD40L融合基因修饰能促进DC活化,增强DC功能,将CD40L胞外段基因与HBV抗原基因融合可能是增强乙肝疫苗免疫效果的有效方法。
Objective To investigate the effects of HBV S - ecdCD40L fusion gene modification on function of dendritic cells（ DC）. Methods The peripheral blood mononuclear cells （PBMCs） from healthy adults were incubated and induced into DC in presence of gran- ulocyte -macrophage colony stimulating factor （GM -CSF） and interleukin -4 （IL -4）. On the fifth day of incubation, DC were trans- feeted with pcDNA3.1 -S -ecdCD40L, pcDNA3.1 -S, pcDNA3.1 ,PBS respectively. DC and the culture supernatants were harvested for detection after 48 hours. The expression level of CD80,CD86 and HLA - DR on DC were detected by flow cytometry. The level of IL - 12 released by DC was analyzed by enzyme - linked immunosorbent assay（ELISA） and the stimulatory capacity of DC in allogenic mixed leu- kocyte reaction （MLR） was tested by CCK -8. Results Modification of DC with HBV S - ecdCD40L fusion gene resulted in activation of DC with up - regulated expression of immuneologieally important cell surface molecules （ CD80, C[）86 and HLA - DR） and pro - in-flammatory cytokines （ IL - 12） , compared with control groups. logeneic T- cell proliferation in vitro. Conclusion Modified enhance its function. Vectors containing HBV S - ecdCD40L therapy. DC modified with HBV S - ecdCD40L were able to stimulate enhanced al- with HBV S - ecdCD40L fusion gene could promote DC＇s activation and fusion gene may be a promising vaccine candidate for chronic hepatitis
出处
《医学研究杂志》
2013年第12期49-53,共5页
Journal of Medical Research
基金
浙江省自然科学基金资助项目(LY12H03003)
