不同途径的供者淋巴细胞输注对移植物抗宿主病的影响

Influence of intra-bone marrow injection of donor lymphocytes upon the development of graft-versus-host disease

目的:观察髓腔内供者淋巴细胞输注(IBM-DLI)对异基因小鼠外周造血干细胞移植(allo-PBSCT)后移植物抗宿主病(GVHD)的影响。方法:雌性C57BL/6小鼠为受鼠,接受全身照射(TBI)预处理后,输注雄性BALB/c小鼠来源的经rhG-CSF动员后的外周造血干细胞,分别经尾静脉(IV)和髓腔内进行DLI,建立异基因GVHD模型,观察移植后小鼠的生存状态和GVHD发生情况,应用流式细胞仪检测受鼠体内嵌合体形成和CD4+CD25+调节性T细胞(Tregs)比例,酶联免疫吸附实验(ELISA)检测白细胞介素4(IL-4)、γ干扰素(IFNγ-)水平。结果:IBM-DLI组的受鼠GVHD发生比例和严重程度较IV-DLI组明显降低(P<0.01);移植后第7天各组受鼠骨髓中供鼠来源的细胞比例均在95%以上;与IV-DLI组比较,脾细胞中Tregs比例在IBM-DLI组明显升高(P<0.01),IBM-DLI组IL-4分泌增多,IFNγ-分泌减少(P<0.01)。结论:与IV-DLI相比,IBM-DLI有利于减轻GVHD的发生,其机制可能与受鼠体内Tregs细胞比例增高以及Th细胞向Th2细胞分化有关。目的:观察髓腔内供者淋巴细胞输注(IBM-DLI)对异基因小鼠外周造血干细胞移植(allo-PBSCT)后移植物抗宿主病(GVHD)的影响。方法:雌性C57BL/6小鼠为受鼠,接受全身照射(TBI)预处理后,输注雄性BALB/c小鼠来源的经rhG-CSF动员后的外周造血干细胞,分别经尾静脉(IV)和髓腔内进行DLI,建立异基因GVHD模型,观察移植后小鼠的生存状态和GVHD发生情况,应用流式细胞仪检测受鼠体内嵌合体形成和CD4+CD25+调节性T细胞(Tregs)比例,酶联免疫吸附实验(ELISA)检测白细胞介素4(IL-4)、γ干扰素(IFN-γ)水平。结果:IBM-DLI组的受鼠GVHD发生比例和严重程度较IV-DLI组明显降低(P<0.01);移植后第7天各组受鼠骨髓中供鼠来源的细胞比例均在95%以上;与IV-DLI组比较,脾细胞中Tregs比例在IBM-DLI组明显升高(P<0.01),IBM-DLI组IL-4分泌增多,IFN-γ分泌减少(P<0.01)。结论:与IV-DLI相比,IBM-DLI有利于减轻GVHD的发生,其机制可能与受鼠体内Tregs细胞比例增高以及Th细胞向Th2细胞分化有关。

Objective：To investigate the effect of donor lymphocyte infusion （DLI） by intra-bone marrow （IBM） or intravenous （IV） on the incidence of graft-versus-host disease（GVHD） after allogeneic peripheral hematopoietic stem cell transplantation （allo-PBSCT）. Methods：Female C57BL/6 mice as recipients received total body irradiation （TBI） on day 0, followed by injection of peripheral hematopoietic stem cells from mobilized male BALB/c with granulocyte-colony stimulating factor （rhG-CSF） ,and DLI was performed via IV or IBM.The extent of GVHD was compared in recipients received allogeneic IBM-DLI with those received IV-DLI. The percentage of donor-derived cells and CD4＋ CD25＋ regulatory T cells （Tregs） was detected by flow cytometry. 14 days after DLI,the levels of IL-4 and interferon （IFN）-γ were tested by ELISA.Results：It was found that the frequency and severity of GVHD were reduced in IBM-DLI compared with that of IV-DLI （P 〈 0.01）. 7 days after transplantation,the percentage of H-2a-positive cells was over 95% in all surviving transplanted mice;and 14 days after transplantation, the percentage of Tregs detected as CD4＋ CD25＋ was significantly higher in recipients treated with IBM-DLI than those treated with IV- DLI（ P 〈 0.01 ）. Compared with that of the recipients in IBM-DLI group, the level of IL-4 was significantly decreased, while the level of IFN-γ were elevated in group IV-DLI （ P 〈 0.01）. Conclusion： IBM-DLI could induce the proliferation of Tregs and the Th polarizing to Th2, resulting in decreasing the incidence and alleviating severity of GVHD after allo-PBSCT.