摘要
目的通过研究VC处理过的DC细胞在过继免疫T细胞培养中是否发挥促进作用,进一步探讨维生素C对于免疫调控的作用机制,同时为制备非特异性CTL提供方法及思路。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VC对DC进行孵育(24 h)后经PBS洗涤,培养5 d后与添加anti-CD3+单抗的T细胞混合培养至第12天,同时设立空白对照组(none DC组、0 mmol/L VC-DC组)。流式细胞仪检测T细胞亚群及细胞因子分泌情况。LDH法检测VC对DC-T细胞非特异性杀伤情况。结果 VC处理的DC细胞可显著刺激T细胞成熟,其CD8+CD28+及CD40L表型有显著升高(61.23%±6.78%,68.87%±4.37%),同时发现Treg细胞明显降低(4.51%±1.22%)。细胞因子分泌情况分析结果显示,vc DC可促进T细胞因子分泌,IFN-γ、IL-2有显著升高(32.29%±3.26%,19.66%±4.37%),IL-4有显著降低(3.36%±1.29%)。杀伤结果显示对不同肿瘤细胞非特异性杀伤有明显增强。结论 VC通过致敏DC进而可激活T细胞向非特异性CTL(CD8+CD28+)分化,提高T细胞因子分泌能力。
The purpose of this study was to illustrate whether vitamin C（VC）- treated DC play promoting rolein T cell culture or not, to further explore the immunoregulation mechanism of VC, and providing methods and ideasfor the nonspecific CTL production. Peripheral blood（about 50 ml） was collected from healthy volunteers, andperipheral blood mononuclear cells were solated with lymphocyte separation medium for DC. 24 hours afterincubation with VC, the DC were washed with PBS and cultured for 5 days, then mixed with anti-CD3＋T cells andcultivated for 12 days. Meanwhile, blank control group（none DC group, 0 mmol/L VC-DC group） was set up. T cellsubgroup and cytokine secretion were detected by flow cytometry; the nonspecific killing of vc DC-T cells to cancerlines was analyzed with LDH method. Data showed that VC-activated DC could significantly stimulate T cellsmaturation, improve the levels of CD8＋CD28＋and CD40 L phenotypes significantly（61.23% ± 6.78%, 68.87% ±4.37%）, and also reduce Treg cells significantly（4.51% ± 1.22%）. The cytokines secreting analysis showed thatVC-DC increased the levels of IFN-γ（32.29%±3.26%） and IL-2（19.66%±4.37%） significantly, but reduced IL-4level（3.36% ± 1.29%） significantly. All result illustrated that vc DC could obvious promote nonspecific CTL（nonspecific CTL） differentiation and T cell factor secretion ability.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2016年第1期19-24,共6页
Immunological Journal
基金
山西省生物治疗示范平台项目(2014091105-0101)
