异基因骨密质来源间充质干细胞对小鼠急性GVHD的防治作用及其机制研究

发布时间：2018-07-06 14:51

本文选题：间充质干细胞 + 造血干细胞移植　；参考：《中国免疫学杂志》2016年11期

【摘要】：目的:明确异基因骨密质来源的间充质干细胞(CB-MSCs)是否能有效防治急性GVHD,并进一步探讨其可能的治疗机制。方法:采用骨密质碎片法从C57BL/6(H-2b)小鼠分离和扩增MSCs。通过构建小鼠allo-HSCT后GVHD模型:雌性供鼠C57BL/6(H-2b)→雄性受鼠BALB/c(H-2d),然后观察CB-MSCs移植对GVHD的影响。本实验小鼠共分为4组:1单纯照射组;2GVHD组;3GVHD~+MSCs组;4正常对照组。在GVHD诱导后不同时间点观察各组小鼠生存率、体重变化和临床GVHD积分;应用ELISA方法检测各组小鼠外周血细胞因子(TNF-α、IFN-γ和IL-4)和趋化因子(CCL5、CXCL9和CXCL10)浓度;应用流式细胞术检测各组小鼠外周血CD4~+CD25~+Foxp3~+调节性T细胞(Treg细胞)百分比和CD3~+T细胞表面趋化因子受体CXCR3、CCR5和CCR7的表达。此外,我们还应用实时定量逆转录PCR(Real-time RT-PCR)方法检测了各组小鼠骨髓中T-bet和GATA-3的mRNA表达水平。结果:CB-MSCs能显著提高GVHD小鼠生存率、减少其体重缺失,并显著降低临床GVHD积分;CB-MSCs对GVHD的防治作用可能与降低外周血TNF-α、IFN-γ、CCL5、CXCL9和CXCL10浓度,增加Treg细胞百分比,下调T细胞表面CXCR3、CCR5的表达和上调CCR7的表达有关。此外,诱导骨髓Th1/Th2平衡向抗炎的Th2极倾斜可能也是CB-MSCs对GVHD发挥治疗作用的机制之一。结论:异基因CB-MSCs显著增加了GVHD小鼠生存率。其治疗机制可能与CB-MSCs能够减少炎性细胞因子和趋化因子的释放,诱导Treg的生成,调控T细胞表面趋化因子受体的表达,以及纠正Th1/Th2的失衡有关。
[Abstract]:Aim: to determine whether allogeneic bone dense mesenchymal stem cells (CB-MSCs) can effectively prevent and treat acute GVHD and explore its possible therapeutic mechanism. Methods: MSCs were isolated and amplified from C57BL / 6 (H-2b) mice by bone mass fragment method. The GVHD model after allo-HSCT in mice was established: female donor C57BL / 6 (H-2b) male recipient BALB / c (H-2d), and then the effect of CB-MSCs transplantation on GVHD was observed. The mice were divided into 4 groups. The survival rate, weight change and clinical GVHD score were observed at different time points after GVHD induction, and the concentrations of peripheral blood cytokines (TNF- 伪, IFN- 纬 and IL-4) and chemokines (CCL5, CXCL9 and CXCL10) were detected by Elisa. The percentage of CD4 ~ CD25 ~ Foxp3- regulatory T cells (Treg cells) and the expression of chemokine receptors CXCR3CCR5 and CCR7 on CD3T cells were detected by flow cytometry. In addition, the expression of T-bet and GATA-3 mRNA in bone marrow of mice in each group was detected by real-time reverse transcription-polymerase chain reaction (Real-time RT-PCR). Results the survival rate of GVHD mice was significantly increased and the weight loss was reduced. The preventive and therapeutic effects of CB-MSCs on GVHD might be related to the decrease of the concentration of CCL5, CXCL9 and CXCL10 in peripheral blood, and the increase of the percentage of Treg cells in GVHD mice, and the effect of CB-MSCs on GVHD might be related to the decrease of the concentration of CCL5, CXCL9 and CXCL10 in peripheral blood. The down-regulation of the expression of CXCR3 and CCR5 on T cells is related to the up-regulation of CCR7. In addition, inducing bone marrow Th1 / Th2 balance to the anti-inflammatory Th2 pole tilt may also be one of the mechanisms of CB-MSCs to play a role in the treatment of GVHD. Conclusion: allogeneic CB-MSCs can significantly increase the survival rate of GVHD mice. The therapeutic mechanism of CB-MSCs may be related to its ability to reduce the release of inflammatory cytokines and chemokines, to induce Treg production, to regulate the expression of chemokine receptors on T cells, and to correct the imbalance of Th1 / Th2.
【作者单位】：河北医科大学第二医院血液科;
【分类号】：R457.7

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