供体TReg细胞对小鼠异基因骨髓移植后GVHD及造血重建影响.docVIP

供体TReg细胞对小鼠异基因骨髓移植后GVHD及造血重建影响 　 作者：杨凯，刘启发，范志平，张钰 【摘要】 　　本研究探讨供体CD4+CD25+调控T细胞(regulatory T cell, TReg)输注对异基因骨髓移植(alloBMT)后小鼠移植物抗宿主病(GVHD)和造血重建的影响。建立C57BL/6→BALB/c小鼠alloBMT模型，体外磁珠分离纯化供鼠外周血CD4+CD25+TReg细胞，接受移植的小鼠随机分为3组，在移植骨髓后6-8小时分别予尾静脉输注CD4+CD25+T细胞、CD4+CD25-T细胞和RPMI 1640培养液（空白对照）。以移植后GVHD表现，肝、脾、小肠组织病理形态、生存期及白细胞(WBC)、血小板(Plt)的重建为观察指标并进行组间比较。结果显示： TReg阳性细胞移植组，TReg阴性细胞移植组和空白对照组小鼠WBCgt;1.0×109/L的时间分别为(8.14±3.26)天、(17.62±5.71)天、(19.81±6.77)天； Pltgt;20.0×109/L的时间分别为(5.29±1.34)天、(8.97±3.44)天、(9.52±3.92)天，TReg阳性细胞移植组WBC和Plt恢复时间比TReg阴性组与空白对照组快(Plt;0.05)。3组小鼠在移植后15天GVHD评分为(1.33±0.58)、(1.80±0.27)、(1.93±0.45)，TReg阳性细胞移植组小鼠与TReg阴性细胞移植组、空白对照组相比差异有统计学意义(Plt;0.05)。移植后(25-30天)的TReg阳性细胞移植组小鼠肝、脾、小肠GVHD病理损害较同期TReg阴性细胞移植组与空白对照组小鼠有一定程度的减轻。3组小鼠移植后平均存活时间分别为(41.45±17.88)天、(18.75±14.39)天和(25.67±16.84)天，TReg阳性细胞移植组小鼠平均存活时间较TReg阴性细胞、空白对照组小鼠延长(Plt;0.05)。结论：小鼠alloBMT后输注供体TReg细胞可以减轻移植后GVHD程度，延长小鼠生存期，促进造血重建。 【关键词】 异基因骨髓移植 CD4+CD25+调控T细胞 移植物抗宿主病 造血重建 　　Influence of Donor TReg Cells on GVHD and Hematopoietic Reconstitution after Allogeneic Bone Marrow Transplantation in Mice Abstract In order to explore the influence of purified donor regulatory T cells (TReg cells) infused after allogeneic bone marrow transplantation (alloBMT) on GVHD and hematopoietic reconstitution of mice, an alloBMT model of C57BL/6→BALB/c mice was established . CD4+CD25+TReg cells were purified through beadmagnetic activated cells seperated from donor mice peripheral blood. The recipient mice were randomly divided into three groups: CD4+CD25+ T cells, CD4+CD25- T cells and RPMI 1640 culture medium. These cells and RPMI 1640 were infused into recipient mice by caudal veins at about 6 to 8 hours after alloBMT respectively. Incidence of GVHD, pathological lesion of liver, spleen, small intestine, survival time and hematopoietic reconstitution in the recipients were observed after alloBMT. The results showed that the time for WBCgt;1.0×109/L was (8.14±3.26)days, (17.62±5.71)days, (19.81±6.77)days and the time for Pltgt;