骨髓间充质干细胞的输注时机对小鼠骨髓移植后急性移植物抗宿主病的影响  

Effect of transplantation opportunity of bone marrow mesenchymal stem cells on acute graft versus host disease after allogeneic bone marrow transplantation

作　　者：薛智文[1] 王存邦[1] 白海[1] 路继红[1] 司志刚[1] 杨小亮[1] 

机构地区：[1]兰州军区兰州总医院血液科全军血液病中心,730050

出　　处：《中华器官移植杂志》2008年第1期38-41,共4页Chinese Journal of Organ Transplantation

基　　金：全军“十一五”杰出人才基金（06J005）

摘　　要：目的探讨骨髓间充质干细胞（MSCs）输注时机对异基因骨髓移植（allo-BMT）后急性移植物抗宿主病（aGVHD）的影响及其机制。方法以Babl／c小鼠为供者，于无菌条件下获取其骨髓，制成MSCs悬液及骨髓细胞悬液。C57BL／6小鼠为受者，经电子直线加速器行全身照射后，分为5组进行allo-BMT。骨髓移植组受者经尾静脉输注骨髓细胞悬液和RPMI 1640培养液各0．2ml；联合移植组受者经尾静脉输注骨髓细胞悬液和MSCs悬液各0．2ml；延迟输注Ⅰ组受者经尾静脉输注骨髓细胞悬液0．2ml，3d后再输注MSCs悬液0．2ml；延迟输注Ⅱ组受者经尾静脉输注骨髓细胞悬液0．2ml，7d后再输注MSCs悬液0．2ml；对照组受者经尾静脉输注RPMI 1640培养液0．4ml。记录受者存活时间及aGVHD的发生情况，检测外周血白细胞、CD4^＋和CD8^＋淋巴细胞数，测定血清γ干扰素（IFN-γ）和白细胞介素4（IL-4）水平，观察死亡小鼠的肝、脾、小肠及皮肤等组织病理学变化。结果对照组受者均于移植后14d内死亡。骨髓移植组于移植14d以后出现aGVHD的表现，且均于23d内死亡。联合移植组aGVHD的发生率为30％，延迟输注Ⅰ组aGVHD的发生率为60％，两组受者的存活时间均长于骨髓移植组。延迟输注Ⅱ组与骨髓移植组间aCVHD发生率和死亡时间的差异无统计学意义。受者的白细胞数均于全身照射后3d降至最低，各移植组白细胞数皆于移植后回升，但骨髓移植组和延迟输注Ⅱ组的白细胞数未能恢复至正常水平，而联合移植组和延迟输注Ⅰ组的白细胞数均于移植后28d恢复正常。输入MSCs者，CD4淋^＋巴细胞数量明显升高，而CD8^＋淋巴细胞数量下降，以联合移植组最为明显。联合移植组的IFN-γ水平明显低于骨髓移植组，而IL-4水平高于骨髓移植组。各组发生aGVHD的小鼠的肝、脾、小肠及皮肤病理改变基本一致。结论MSCs与骨�Objective To investigate the effect and the mechanism of bone marrow mesenchymal stem cells （MSCS） injected in different time points on acute graft versus host disease （aGVHD） after allogeneic bone marrow transplantation （alIo-BMT）. Methods Both bone marrow cells and cultured MSCS obtained from donor mice Babl/c were transplanted into the recipient mice C57BL/6 after irradiation by electron linear accelerator. In the alIo-BMT group, the mice were transfused with bone marrow cells alone through tail vein, those in the co-transplantation group with bone marrow cells and MSCS simultaneously, those in 3 days interval group and 7 interval group with MSCS at 3rd and 7th day after bone marrow cells transfusion. The survival time after alIo-BMT was recorded, and the general manifestation and pathological changes of aGVHD and hematopoietic reconstitution in recipient mice were observed. The effects of MSCS on the quantity of CD4^＋ and CD8^＋ T cells in vivo after allo- BMT were evaluated by flow cytometry. The serum levels of both IFN-γ and IL-4 were determined by ELISA. Results All mice in the control group died at 14th day. Allo-BMT mice suffered aGVHD at 14th day and died at 23rd day after the transplantation. In co-transplantation group and 3 days interval group 30% and 60% mice suffered aGVHD respectively, and both group mice had longer survival time than allo-BMT group. In 7 days interval group the mice in aGVHD incidence and survival time were as same as allo-BMT mice. White blood cells were lowest at 3rd day after irradiation, and the cells in cotransplantation group and 3 days interval group got normal at 28th day, but in allo-BMT and 7 days interval groups the cells get no recovery. In co-transplantation group the mice had significantly higher CD4^＋ cells and lower CD8^＋ cells, and lower IFN-γ and higher IL-4 than in allo-BMT group. Conclusion Co-transplantation of hone marrow cells with MSCS from the same donor mice can alleviate aGVHD and the effect is most satisfactory if both BM and MSCS

关 键 词：间质干细胞 骨髓移植 移植物抗宿主病 时间 

分 类 号：R457.7[医药卫生—治疗学] R329.2[医药卫生—临床医学]