中国免疫学杂志
中國免疫學雜誌
중국면역학잡지
Chinese Journal of Immunology
2015年
10期
1324-1328
,共5页
贾原%李文丽%李芳%赵晴%张俊萍
賈原%李文麗%李芳%趙晴%張俊萍
가원%리문려%리방%조청%장준평
维生素C%树突状细胞%表面共刺激分子%CD40
維生素C%樹突狀細胞%錶麵共刺激分子%CD40
유생소C%수돌상세포%표면공자격분자%CD40
Vitamin C%Dendritic cell ( DC)%Co-stimulating molecular%CD40
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。
目的：探討維生素C（VC）對樹突狀細胞（DC）的影響，併檢測DC 細胞成熟情況，為快速製備DC 疫苗提供一種可行方法及思路。方法：採集健康誌願者外週血50 ml，用淋巴細胞分離液分離外週血PBMC，貼壁後穫得單覈細胞。用不同濃度VC 對DC 進行孵育（24 h），後經PBS 洗滌，併設立空白對照組（V0）。流式細胞儀檢測DC 錶麵共刺激分子CD80／86及HLA－DR，CD40錶達情況。通過設置VC 濃度梯度及時間梯度，攷察VC 對DC 刺激的最佳濃度及最佳成熟時間，同時分析VC 促進DC 成熟的原因。結果：VC 可顯著刺激DC 細胞成熟，其CD80／86錶達較未添加VC 組有明顯提升。且本研究顯示VC 刺激DC 的最佳濃度為1 mmol／L，噹DC 培養至第三天時，VC 組CD80／86錶達率為（78．6±4．6）％，而空白對照組V0僅為（34．1±5．7）％；DC 錶麵HLA－DR 錶達：VC（56．8±4．4）％，空白對照組V0為（25．4±4．7）％，兩組差異有統計學意義，P＜0．05；進而我們攷察瞭VC 對DC 細胞CD40、CD40L 錶達情況，結果顯示，VC 2．5 mmol／L 組CD40錶達率最高可達（59．3±3．7）％，V0組僅為（11．1±2．4）％，說明VC 可顯著調節DC 錶麵CD40的錶達。而CD40L 錶達調節併未體現。熒光顯微鏡結果顯示VC－DC 抗原捕穫能力顯著提升。結論：VC 可顯著調控DC 成熟，併可能通過上調CD40進而促進CD80／86及HLA－DR 的錶達，噹VC 濃度為1 mmol／L 時，對DC調控作用最彊。
목적：탐토유생소C（VC）대수돌상세포（DC）적영향，병검측DC 세포성숙정황，위쾌속제비DC 역묘제공일충가행방법급사로。방법：채집건강지원자외주혈50 ml，용림파세포분리액분리외주혈PBMC，첩벽후획득단핵세포。용불동농도VC 대DC 진행부육（24 h），후경PBS 세조，병설립공백대조조（V0）。류식세포의검측DC 표면공자격분자CD80／86급HLA－DR，CD40표체정황。통과설치VC 농도제도급시간제도，고찰VC 대DC 자격적최가농도급최가성숙시간，동시분석VC 촉진DC 성숙적원인。결과：VC 가현저자격DC 세포성숙，기CD80／86표체교미첨가VC 조유명현제승。차본연구현시VC 자격DC 적최가농도위1 mmol／L，당DC 배양지제삼천시，VC 조CD80／86표체솔위（78．6±4．6）％，이공백대조조V0부위（34．1±5．7）％；DC 표면HLA－DR 표체：VC（56．8±4．4）％，공백대조조V0위（25．4±4．7）％，량조차이유통계학의의，P＜0．05；진이아문고찰료VC 대DC 세포CD40、CD40L 표체정황，결과현시，VC 2．5 mmol／L 조CD40표체솔최고가체（59．3±3．7）％，V0조부위（11．1±2．4）％，설명VC 가현저조절DC 표면CD40적표체。이CD40L 표체조절병미체현。형광현미경결과현시VC－DC 항원포획능력현저제승。결론：VC 가현저조공DC 성숙，병가능통과상조CD40진이촉진CD80／86급HLA－DR 적표체，당VC 농도위1 mmol／L 시，대DC조공작용최강。
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<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.
