临床儿科杂志 ›› 2015, Vol. 33 ›› Issue (12): 1057-.doi: 10.3969 j.issn.1000-3606.2015.12.014

• 实验研究 • 上一篇    下一篇

1,25- 二羟维生素D3 对脐血单核细胞来源树突状细胞刺激T 细胞增殖能力的影响

李海园,唐征,王翼,施婴婴,刘志伟   

  1. 上海交通大学医学院附属国际和平妇幼保健院新生儿科( 上海 200030)
  • 收稿日期:2015-12-15 出版日期:2015-12-15 发布日期:2015-12-15
  • 通讯作者: 刘志伟 E-mail:liuzhiwei@hotmail.com

Effects of 1,25-dihydroxyvitamin D3 on the stimulating ability of cord blood monocytes-derived dendritic cells to T cells proliferation

LI Haiyuan, TANG Zheng, WANG Yi, SHI Yingying, LIU Zhiwei   

  1. Department of Neonatology, International Peace Maternity and Child Health Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200030, China
  • Received:2015-12-15 Online:2015-12-15 Published:2015-12-15

摘要: 目的 探讨1,25- 二羟维生素D3 [1,25(OH)2D3] 对脐血单个核细胞来源树突状细胞(DCs) 刺激T 细胞增殖能力的影响。方法 无菌条件下采集脐血,体外分离获得脐带血单个核细胞,经重组人粒细胞巨噬细胞集落刺激因子(rhGM-CSF)、重组人白介素-4(rhIL-4)和重组人肿瘤坏死因子-α(rhTNF-α)培养8 d 诱导分化为DCs,予1,25(OH)2D3(10 nmol/L) 干预。流式细胞术检测DCs 表面分子的表达情况,经CD3 磁珠阳性分选后获得T 细胞,并以羧基荧光素二醋酸盐琥珀酰亚胺酯(CFSE)标记,用流式细胞术检测DCs 刺激自体T 细胞增殖的能力和T 淋巴细胞各亚群的增殖情况;ELISA 法检测混合培养上清中细胞因子水平。结果 1,25(OH)2D3 干预组DCs 刺激同种同体CD3+T 细胞增殖率低于对照组(61.6±8.23 对75.21±6.21),且CD3+CD4+ 和CD3+CD8+T 细胞亚群的增殖率均降低(59.24±10.22 对74.54±6.89,47.06±12.10 对59.97±9.11),差异均有统计学意义(P 均<0.05)。DCs 与同种同体T 淋巴细胞混合培养后收集上清液检测细胞因子,1,25(OH)2D3 干预组IL-12 和IFN-γ 浓度均低于对照组(78.84±11.8 对99.06±9.07,9.76±2.75对22.45±2.6),而IL-4 则高于对照组(49.45±2.84 对35.75±1.89),差异均有统计学意义(P 均<0.05)。结论 1,25(OH)2D3可抑制脐血单个核细胞来源DCs 刺激自体CD3+CD4+ 和CD3+CD8+T 淋巴细胞增殖反应,且诱导T 细胞免疫应答呈Th2倾向。

Abstract:  Objective To explore the effects of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) on the stimulating ability of cord blood monocytes-derived dendritic cells (DCs) to T cells proliferation. Methods Umbilical cord blood was collected in aseptic condition. Cord blood monocytes were isolated and cultured for 8 days in the presence of rhGM-CSF, rhIL-4 and rhTNF-α for inducing to differentiate DCs, then intervened by 1,25(OH)2D3 (10 nmol/L). The expressions of differentiation and maturation markers on dendritic cells were determined by flow cytometry. Mixed lymphocyte reaction (MLR) was used and T cells were stained with CFSE to observe the stimulating ability of DCs to T cells proliferation by flow cytometry. Cytokines in the culture medium of MLR were measured by ELISA. Results 1,25(OH)2D3 treated-DCs stimulated less CD3+ T cells to proliferate than control group (61.6±8.23 vs. 75.21±6.21, P<0.01). Moreover, lower proliferation of both CD3+CD4+ and CD3+CD8+ T cells was observed in intervention group (59.24±10.22 vs. 74.54±6.89, P<0.05; 47.06±12.10 vs 59.97±9.11, P<0.05) . The IL-12 and IFN-γ levels in the culture medium of MLR in intervention group were significantly lower than control group (78.84±11.8 vs .99.06±9.07, P<0.05; 9.76±2.75 vs. 22.45±2.6, P<0.05), however, the IL-4 level in intervention group were significantly higher than control group (49.45±2.84 vs. 35.75±1.89, P<0.01). Conclusions 1,25(OH)2D3 inhibits the capability of DCs to activate CD3+CD4+ and CD3+CD8+ T cells, and instructs DCs to induce Th2 bias.