摘要
目的探讨脂质体介导的基因转染对人未成熟树突状细胞(imDC)表型特征及免疫学功能的影响。方法贴壁法分离人脐血来源的单核细胞,用重组人粒细胞巨噬细胞集落刺激因子和重组人白细胞介素4将其诱导分化为imDC后,进行形态学观察和免疫表型鉴定,并将其分为转染组和对照组。转染组将pEGFP-N1质粒通过脂质体转染imDC；对照组不作特殊处理。流式细胞仪检测此法的转染率和两组细胞表面分子[CD86、CD83、人类自细胞DR抗原(HLA-DR)等]的表达率；混合淋巴细胞反应(MLR)检测imDC在转染前后刺激未致敏T淋巴细胞增殖的能力。结果人脐皿来源的imDC形态结构和表面标志符合文献报道的典型特征。脂质体介导的基因转染法转染率约在10%左右。转染组CD86、CD83和HLA-DR的表达率分别为(12±6)%、(8．6±2．3)%和(71±7)%；对照组分别为(13±6)%、(9．1±3．8)%和(72±8)%,两组比较,差异无统计学意义(P＞0．05)。MLR提示imDC转染后其免疫刺激功能较转染前无明显变化(P＞0．05),刺激指数＜2。结论脂质体介导转染imDC后不影响其成熟特性,但转染率不高。
Objective To investigate the influence of liposome induced gene transfection on the phenotypic characteristics and immune function of human immature dedritic cells （imDC）. Methods Monocytes were isolated from human cord blood, and they were differentiated into imDC by rhGM-CSF and rhlL-4 induction. Then the morphologic observation and immune phenotypic identification were performed in imDCs. imDCs were divided into transfection （T, with liposome transfection of pEGFP vector） and control （ C, without transfection） groups. The transfection rate and expression of cell maturation marker （ CD83, CD86 and HLA-DR） were determined with flow cytometry, and the proliferation of non-sensitized T lymphocyte before and after transfection was determined with allogenetic mixed leukocyte reaction （MLR）. Results imDC derived from human cord blood cells had typical appearance and surface markers consistent to what reported in the literature. The expression rates of CD86,CD83 and LA-DR in T group were （12 ±6）% , （8.6 ± 2.3）% and （71 ±7）% , respectively, which exhibited no difference compared with those in C group （13 ± 6）%,（9.1 ±3.8）% and （72 ±8）%,（ P 〉0.05）. MLR results indicated that there was no obvious change in the immune stimulation function of imDC after transfection （ P 〉 0.05 ） , with stimulation index lower than 2. Conclusion There is no change in maturation of imDC after liposome transfection, but the transfection efficiency needs to be elevated.
出处
《中华烧伤杂志》
CAS
CSCD
北大核心
2006年第3期203-206,共4页
Chinese Journal of Burns
基金
国家自然科学基金(30271341)
关键词
树突细胞
转染
脂质体
免疫表型分型
胎血
Dendritic cells
Transfection
Liposomes
Immunophenotyping
Fetal blood
