异基因造血干细胞移植后细胞嵌合状态动态监测的临床意义

Clinical significance of dynamic monitoring of cell chimerism following allogeneic hematopoietic stem cell transplantation

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人、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，存移植后定期采集所有患者的外周m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（＋21d）达到完全供者嵌合状态。＋7dT淋巴细胞供者嵌合比例（DC）≥70％和＋14dT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渊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 allogenieic hematopoietic stein 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 lymphoeytes 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 ehilnerism 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 chimerisin（CDC） with a median on day ＋ 21. Patients whose T lymphoeytes donor chimerism was more than 70% on day ＋7 anti 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 lymphoeytes were observed hefore molecular or hematological relapse occurred. Conclusion Serial anti 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 allo- HSCT.