摘要
目的体外诱导CD8+T细胞,以期诱导分化出一种IL-10分泌性CD8+调节性T细胞,为用细胞疗法进行自身免疫性疾病的预防及治疗奠定基础。方法以tiger转基因小鼠(以其GFP+细胞代表IL-10+细胞)为实验动物,流式细胞仪分选幼稚CD8+T细胞,然后在细胞培养体系中加入ConA或抗CD3以及IL-2,IL-4加或不加,培养48 h对细胞进行诱导分化,继而,细胞继续扩大培养,至96 h时,加入佛波酯和离子霉素对细胞进行再刺激6 h,流式检测CD8+T细胞GFP的产生情况。同时,为验证IL-4对IL-10分泌性CD8+T细胞的诱导分化作用,在培养体系中加入抗IL-4,流式检测CD8+T细胞IL-10的表达情况。结果经流式细胞仪检测幼稚CD8+T在未经诱导分化时不产生IL-10;而在ConA或抗CD3的刺激及IL-4的诱导下,幼稚CD8+T可分化为IL-10分泌性细胞,且其比例分别占CD8+T细胞的31.16%(ConA组)和32.20%(抗CD3组);培养体系中加入抗IL-4后,IL-10分泌性CD8+T细胞的分化被抑制,其比例降为11.2%。结论 IL-10分泌性CD8+T细胞可由CD8+T诱导分化而来,IL-4是诱导IL-10产生的关键细胞因子。
Objective To study a way to induce nave CD8^＋ T cell differentiation into IL-10-secreting CD8^＋ T cells in vitro,in order to find a new subset of CD8^＋ regulatory T cell（Treg）,so as to provide the basis for using cell therapy for disease of self-immunity. Methods Nave CD8^＋T cells（CD62L^hiCD44^lo） from tiger mice（in which green fluorescent protein positive（GFP^＋） cells are used to indicate cells expressing IL-10） were sorted,mixed with DCs and stimulated with Con A ＋ IL-2 or activated with soluble anti-CD3^＋ IL-2 in the absence or presence of IL-4 for 48 h and then expanded for 48 h. Cells were then restimulated for 6 h with phorbol-12-myristate-13-acetate（PMA） and ionomycin（ION） and stained.The GFP^＋CD8^＋T cells were detected by flow cytometry. In the parallel experiment,anti-IL-4 was added to the medium to define the role of IL-4. Results There was no IL-10 expression in nave CD8^＋T cells. But after stimulation of PMA and ionomycin,IL-4 induced a significantly higher number of GFP^＋T cells under Con A and antiCD3 conditions（the ratios of CD8^＋GFP^＋T cells were 31. 16% and 32. 2% respectively）;while after addition of anti-IL-4,the differentiation of CD8^＋GFP^＋T cells was suppressed,the ratio of CD8^＋GFP^＋T cells was only 11. 2%. Conclusion IL-4 plays a critical role in inducing IL-10-secreting CD8^＋T cells.
出处
《解放军预防医学杂志》
CAS
2014年第4期296-298,共3页
Journal of Preventive Medicine of Chinese People's Liberation Army
基金
天津市自然科学基金面上项目(No.14JCYBJC24500)
