Human Umbilical Cord Blood-derived Stromal Cells Suppress Xenogeneic Immune Cell Response In Vitro
Human Umbilical Cord Blood-derived Stromal Cells Suppress Xenogeneic Immune Cell Response In Vitro
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DOI：
10.3325/cmj.2009.50.351
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发表时间：
2009-08-01
影响因子：
1.9
通讯作者：
Wang, Qing-yu
中科院分区：
文献类型：
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作者：
Hao, Lei;Zhang, Cheng;Wang, Qing-yu
目的探讨人脐血源基质细胞(HUCBDSC)的免疫学特性及其对异种免疫细胞的影响。方法采用流式细胞术检测新鲜分离和冷冻保存的hUCBDSC的免疫表型。用植物血凝素A(PHA)或树突状细胞刺激异种脾T细胞，不加或不加hUCBDSC。孵育7d后，用细胞计数试剂盒8检测T细胞增殖情况。用流式细胞术检测在无或有hUCBDSCs存在的情况下，经PHA激活的T细胞中的凋亡细胞和CD4(+)CD25(+)Foxp3(+)调节性T细胞(Tregs)的比例。结果新鲜分离的人脐血树突状细胞的免疫分子表达水平分别为：人白细胞抗原-I(84.1+/-2.9%)、人类白细胞抗原-II(1.6+/-0.3%)、CD80(0.8+/-0.1%)、CD86(0.8+/-0.1%)、CD40(0.6+/-0.1%)、CD40(0.5+/-0.1%)、CD80(0.8+/-0.1%)、CD40(0.6+/-0.1%)、CD40(0.5+/-0.1%)、CD40(0.5+/-0.1%)、CD80(0.8+/-0.1%)、CD80(0.8+/-0.1%)。不受冷冻保存的影响。在hUCBDSC存在下，T细胞的增殖明显低于阳性对照。共培养导致10倍的增长(从1.2+/-0.3%增加到12.1+/-1.4%，P<；0.001)CD_4~+、CD_(25)~+、FOX_3~(+)调节性T细胞和成熟树突状细胞逆转，主要组织相容性复合体II分子表达下调(49.3%vs 25.9%，P=0.001)，CD_(80)(47.2%vs 23.3%，P=0.001)，CD_(86)(40.6%vs 25.1%，P=0.002)。结论hUCBDSCs具有较低的免疫原性和免疫调节作用。HUCBDSCs对异种T细胞增殖的抑制作用可能与成熟树突状细胞逆转和Treg比例增加有关，而与细胞凋亡无关。
Aim To explore immunological properties of human umbilical cord blood-derived stromal cells (hUCBDSC) and their effect on xenogeneic immune cells in vitro.Methods Immunological phenotype of freshly isolated and cryopreserved hUCBDSCs was evaluated by flow cytometry. Xenogeneic splenic T-cells were stimulated by phytohemaglutinin A (PHA) or dendritic cells in the absence or presence of hUCBDSCs. T-cell proliferation was measured by cell counting kit-8 after 7-day incubation. The proportion of apoptotic cells and CD4(+)CD25(+)Foxp3(+) regulatory T-cells (Tregs) was determined in T-cells activated by PHA in the absence or presence of hUCBDSCs by flow cytometry. Phenotype of dendritic cells, cultured alone or with hUCBDSCs, was analyzed by flow cytometry.Results Levels of immune molecule expression on freshly isolated hUCBDSCs were as follows: human leukocyte antigen-I (HLA-I) (84.1 +/- 2.9%), HLA-II (1.6 +/- 0.3%), CD80 (0.8 +/- 0.1%), CD86 (0.8 +/- 0.1%), CD40 (0.6 +/- 0.1%), and CD40L (0.5 +/- 0.1%), which was not influenced by cryopreservation. T-cell proliferation in the presence of hUCBDSCs was significantly lower than that of positive control. The coculture led to a 10-fold increase (from 1.2 +/- 0.3% to 12.1 +/- 1.4%, P < 0.001) in the proportion of CD4(+)CD25(+)Foxp3(+) regulatory T-cells (Tregs) and a reversion of mature dendritic cells, as indicated by the down-regulation of major histocompatibility complex (MHC)-II molecule (49.3% vs 25.9%, P = 0.001), CD80 (47.2% vs 23.3%, P = 0.001), and CD86 (40.6% vs 25.1%, P = 0.002). When subjected to annexin V binding and propidium iodide uptake assay, the hUCBDSCs did not show the ability to induce apoptosis of xenogeneic T-cells.Conclusion These results demonstrate low immunogenicity and immunomodulation effect of the hUCBDSCs. Reversion of mature dendritic cells and increase in Treg proportion, but not cell apoptosis, can possibly contribute to the suppression of xenogeneic T-cell proliferation by the hUCBDSCs.