摘要
目的初步探讨成人脂肪源间充质干细胞(adultadiposetissue-derivedmesenchymalstemcells,AMSC)治疗急性移植物抗宿主病(acutegraft-versus-hostdisease,aGVHD)的分子机制。方法3例行异基因造血干细胞移植术后发生aGVHD患者,以每公斤体质量2×106个细胞剂量静脉输注AMSC;应用RT-PCR扩增患者AMSC使用前后外周血单个核细胞的TCRVβ24个亚家族的CDR3,了解患者TCRVβT细胞的分布情况;应用尼龙毛柱分离外周血T淋巴细胞再经CD8磁珠分选出CD8+T淋巴细胞,应用流式细胞术检测发生aGVHD患者使用AMSC前后外周血CD8+T细胞亚群的变化。结果患者发生aGVHD时,有TCRVβ3及其他亚家族基因表达,输注AMSC后,GVHD得以有效控制,Vβ不表达;当患者GVHD复发时,Vβ3基因又表达,治疗后不表达;与输注AMSC前相比,输注AMSC后,CD8+T细胞中的CD8+CD28-亚群显著上调(P<0.05),同时,患者的aGVHD得以有效控制。结论AMSC治疗aGVHD的分子机制可能与其抑制TCRVβ亚家族基因表达有关,TCRVβ3可能是AMSC作用的靶基因;同时,AMSC治疗aGVHD的作用机制可能与其上调CD8+CD28-T细胞亚群有关,CD8+T细胞可能是AMSC作用的靶细胞。
Objective To study the molecular mechanism of adult adipose tissue-derived mesenchymal stem cells （AMSC） in treatment of patients with acute graft-versus-host disease （aGVHD）. Methods A dose of 2 × 10^6 AMSC/kg wt was given intravenously to the patients with aGVHD. The distribution of TCR Vβ T repertoire was determined with amplification of the CDR3 of TCR Vβ 24 subfamily genes by using RT-PCR in 3 cases with aGVHD following allogeneic hematopoietic stem cell transplantation. In addition ,T cells were harvested by using nylon column and CD8^＋ T cells sorted by magnetic beads and the proportion of CD8^＋CD28^- T cells was assayed by fluorescence-activated cell sorter（FACS）. Results It was found that there were expressions of TCR Vβ3 gene and other subfamily genes when aGVHD was obvious,but they were not detected after AMSC treatment with remission of GVHD. While during relapse of the GVHD ,expressions of Vβ3 gene reappeared,but disappeared after treatment. The data of FACS indicated that in the CD8^＋ T cells,CD8^＋CD28^- T cells were up-regulated significantly after AMSC transplantation （P 〈 0.05 ）. At the same time, a complete remission of aGVHD disease was observed. Conclusion The molecular mechanism for the treatment with AMSC in aGVHD might be correlated with the inhibition of TCR Vβ subfamily gene expressions. The TCR Vβ3 gene might be the target gene for AMSC and also might be correlated with the up-regulation of CD8^＋CD28^- T cells in vivo.
出处
《生物医学工程与临床》
CAS
2008年第4期290-294,共5页
Biomedical Engineering and Clinical Medicine
基金
河南省杰出青年基金(0612000900)
河南省医学科技创新人才工程项目(200590)
