目的 研究异基因造血干细胞移植(allo-HSCT)后各系细胞嵌合状态与移植物植入、急性移植物抗宿主病(aGVHD)、移植物被排斥和疾病复发的关系.方法 对65例患者进行allo-HSCT,在移植后定期采集所有患者的外周血和骨髓.用流式细胞术分选了65例患者的CD3+T淋巴细胞,52例分选了CD3-CD56+CD16+NK细胞,32例分选了CD15+粒细胞,20例分选了CD19+B淋巴细胞.进行PCR扩增短串联重复序列检测各系细胞嵌合状态.结果 移植后NK细胞早期植入比例(55.5%)最高,T细胞最晚(+21 d)达到完全供者嵌合状态.+7 d T淋巴细胞供者嵌合比例(DC)≥70%和+14 d T淋巴细胞DC≥95%属aGVHD发生的高危患者.除急性淋巴细胞白血病外,出现移植物被排斥的分子生物学征象者和疾病复发者,都以T淋巴细胞供者嵌合状态下降为主.在过继免疫治疗中,动态检测嵌合状态可以判断疗效和指导进一步的治疗.结论 allo-HSCT后T淋巴细胞嵌合状态动态检测可以早期预测发生aGVHD的高危患者、判断移植物植入、发现移植物被排斥和疾病复发并指导免疫调节治疗的时机和疗效.
Objective To evaluate the relationship of chimerism status of cell subsets with engraftment, occurrence of acute graft versus host disease (aGVHD), graft rejection and disease relapse after aliogenieic hematopoietic stem cell transplantation (allo-HSCT). Methods Chimerism status in peripheral blood (PB) and bone marrow (BM) of 65 patients received allo-HSCT were monitored at regular intervals posttransplant. Fluorescence-activated cell sorter (FACS) was used to sort CD3+ T lymphocytes in 65 cases, CD3- CD56+ CD16+ NK cells in 52 cases, CD15+ granulocytes in 32 cases and CD19+ B lymphocytes in 20 cases post transplants. The chimerism status of different lineage cells was analyzed by polymerase chain reaction amplification of short tandem repeats (PCR-STR). Results On day + 7, NK-cells donor chimerism (DC 55.5%)was higher than other cell subsets. T lymphocyte was the latest one to reach complete donor chimerism(CDC) with a median on day + 21. Patients whose T lymphocytes donor chimerism was more than 70% on day +7 and more than 95% on day + 14 had a high risk for acute aGVHD. In all cases except those with ALL, the decreased DC of T lymphocytes were observed before molecular or hematological relapse occurred. Conclusion Serial and quantitative T cell chimerism analysis provides a reliable and rapid screening method for the early detection of engraftment, graft rejection, disease relapse and occurrence of aGVHD, therefore, is a prognostic tool to identify patients at high risk of aGVHD and disease relapse following alloHSCT.