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  

Allogeneic compact bone-derived mesenchymal stem cells attenuates severity of GVHD in a murine bone marrow transplantation model

作　　者：乔淑凯[1] 郭晓楠[1] 张雅丽[1] 任金海[1] 王颖[1] 张静楠[1] 李杨[1] 

机构地区：[1]河北医科大学第二医院血液科,石家庄050000

出　　处：《中国免疫学杂志》2016年第11期1672-1677,共6页Chinese Journal of Immunology

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。Objective： To determine whether allogeneic murine compact bone derived-mesenchymal stem cells（ CB-MSCs）could prevent the development of GVHD and investigate the potential mechanism. Methods： MSCs were isolated and amplification from C57 BL / 6（ H-2b） mice by bone fragments method. The effects of CB-MSCs on GVHD were tested by using an established murine model of C57 BL / 6（ H-2b） →BALB / c（ H-2d）. The mice were divided into 4 groups： 1simple irradiation group; 2GVHD group; 3GVHD ＋MSCs group; 4the normal control group. Survival rates,body weight change and clinical GVHD scores were assessed after GVHD induction. Mechanistically,concentrations of cytokines（ TNF-α,IFN-γ and IL-4） and chemokines（ CCL5,CXCL9 and CXCL10） from peripheral blood in the recipient mice were measured at different time point post-transplantation. CD4＋CD25＋Foxp3＋regulatory T cell（ Treg） percentage,CCR5,CXCR3 and CCR7 expression on CD3＋T cells,and T-bet and GATA-3 mRNA levels were also evaluated at different time point post-transplantation. Results： CB-MSCs significantly attenuated the severity of GVHD and increased survival rate of mice. A higher Treg percentage,reduction of TNF-α,IFN-γ,CCL5,CXCL9 and CXCL10 levels,down-regulation of CXCR3 and CCR5,as well as up-regulation of CCR7,were observed in MSCs infused mice. Also,the prophylactic effect of CB-MSCs was associated with a shift of Th1 / Th2 balance toward anti-inflammatory Th2 polarization. Conclusion： Allogeneic CB-MSCs may offer a novel prophylactic approach for GVHD after allo-HSCT.

关 键 词：间充质干细胞 造血干细胞移植 移植物抗宿主病 调节性T细胞 趋化因子受体 

分 类 号：R392.4[医药卫生—免疫学]