摘要
目的:探讨维生素C(VC)对树突状细胞(DC)的影响,并检测DC细胞成熟情况,为快速制备DC疫苗提供一种可行方法及思路。方法:采集健康志愿者外周血50 ml,用淋巴细胞分离液分离外周血PBMC,贴壁后获得单核细胞。用不同浓度VC对DC进行孵育(24 h),后经PBS洗涤,并设立空白对照组(V0)。流式细胞仪检测DC表面共刺激分子CD80/86及HLA-DR,CD40表达情况。通过设置VC浓度梯度及时间梯度,考察VC对DC刺激的最佳浓度及最佳成熟时间,同时分析VC促进DC成熟的原因。结果:VC可显著刺激DC细胞成熟,其CD80/86表达较未添加VC组有明显提升。且本研究显示VC刺激DC的最佳浓度为1 mmol/L,当DC培养至第三天时,VC组CD80/86表达率为(78.6±4.6)%,而空白对照组V0仅为(34.1±5.7)%;DC表面HLA-DR表达:VC(56.8±4.4)%,空白对照组V0为(25.4±4.7)%,两组差异有统计学意义,P<0.05;进而我们考察了VC对DC细胞CD40、CD40L表达情况,结果显示,VC 2.5 mmol/L组CD40表达率最高可达(59.3±3.7)%,V0组仅为(11.1±2.4)%,说明VC可显著调节DC表面CD40的表达。而CD40L表达调节并未体现。荧光显微镜结果显示VC-DC抗原捕获能力显著提升。结论:VC可显著调控DC成熟,并可能通过上调CD40进而促进CD80/86及HLA-DR的表达,当VC浓度为1 mmol/L时,对DC调控作用最强。
Objective：To study the influence of vitamin C （ VC） on dendritic cells （ DC） ,and detect DC maturation,to provide a feasible method and thought for quickly preparating DC vaccines.Methods：Collected the peripheral blood （about 50 ml） from healthy volunteers,and isolated peripheral blood mononuclear cells with lymphocyte separation medium and obtain DC.With stimulating with different concentrations of VC for （24 h）,then washed with PBS,and set up blank control group （V0）.The expression of DC surface co-stimulating molecules CD80/86 and HLA-DR, CD40 was detected by flow cytometry.By setting the concentration gradient and time gradient, exciting optimal concentration and stimulating time of VC on DC, and analyzed the reasons of VC promoting DC maturation.Results：VC could effectively stimulate DC,CD80/86 expression had significantly increased contrast to the blank control group （V0）.And the experiments show that VC’s best stimulating concentration was 1 mmol/L,and on the third day,the CD80/86 expression rate of VC group was （78.6±4.6） %,and blank control group V0 was （34.1±5.7） %.DC surface HLA-DR expression：VC （56.8± 4.4） %,blank control group V0 （25.4 ±4.7） %,the difference between two groups was statistically significant,P＆lt;0.05.CD40 and CD40L expression and results show that VC 2.5 mmol/L group of CD40 expression rate up to （59.3±3.7） %,while V0 group was only （11.1 ±2.4） %,that illustrate VC could significantly regulate CD40 expression on DC surface,but CD40L not reflect.Fluorescence mi-croscope results showed that DC’ s antigen catching ability was also significantly promoted.Conclusion：VC can significantly regulate DC maturity,and may up regulate CD40,thus promoting the express of CD80/86 and HLA-DR.When the concentration is 1 mmol/L,VC expresses the strongest regulation function on DC.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2015年第10期1324-1328,共5页
Chinese Journal of Immunology
基金
山西省生物治疗示范平台项目资助(No.2014091105-0101)
