荧光原位杂交技术在判断异基因造血干细胞移植预后中的应用

Application of fluorescence in situ hybridization in the prognostic estimation of allogeneic hematopietic stem cell transplantation

目的 研究荧光原位杂交（FISH）技术在判断异基因造血干细胞移植（allo-HSCT）预后中的意义.方法 收集异基因外周血干细胞移植、骨髓移植、脐血移植及多种干细胞联合移植标本63例,收集条件为融合基因（BCR/ABL、AML1/ETO）阳性表达,供受者性别不同或同时满足以上2项条件者,分为同性别间干细胞移植和不同性别间干细胞移植两组：同性别间干细胞移植者20例,应用相应的基因探针进行检测;不同性别间干细胞移植者应用相应的基因探针及XY染色体探针联合检测.结果 43例异性allo-HSCT的受者中,25例供、受者骨髓嵌合率在99%以上;13例早期嵌合率偏低（82%~98%）,在随访过程中逐渐上升至99%以上,微量残留病灶（MRD）阴性,移植后原病均未复发;3例嵌合率由99%~100%逐渐下降,MRD比例上升,出现血液学复发.20例供、受者性别相同的allo-HSCT受者中,15例移植后未检测到原来的异常基因;2例检测到微量残留病灶,经免疫抑制剂减量、供者淋巴细胞输注等治疗后降为1%,目前仍在完全缓解中;3例在移植后1~4个月时出现原来的基因异常,复查骨髓为原病复发.结论 FISH技术简便、快速、准确性高、特异性强,对异基因干细胞移植预后判断具有重要意义.

Objective To investigate the significance of fluorescence in situ hybridization（FISH） in prognostic estimation of allogeneic hematopietic stem cell transplantation（allo-HSCT）. Methods 63 cases of samples were collected from patients with peripheral blood stem cell transplantation, bone marrow transplantation, umbilicus stem cell transplantation or combined transplantation of multiple stem cell. All enrolled patients were positive with fusion genes （BCR/ABL, AML1/ETO）, with gender difference with cor responding donors,or be fit for both of the two criteria,and were divided into two groups of same sex and different sex allo-HSCT, according to the sex of donors and patients. 20 cases were enrolled in group of same sex allo-HSCT and were detected by using corresponding gene probes,and 43 cases in group of different sex allo HSCT were detected by using corresponding gene probe and combined XY chromosome probes. Results Among the 4,3 cases of the opposite sex allo-HSCT, 25 cases of chimerism ratio of bone marrow of donors and recipients were higher than 99% , chimerism ratio of 13 cases, with low early chimerism ratio （82%- 98% ）, gradually increased to more than 99% during follow-up, without minimal residual disease（MRD） and relapsed original disease after transplantation, and ehimerism rate of 3 cases decreased from 99 %- 100 G, with hematological relapse and increased MRD positive rate. Among 20 cases of same sex allo-HSCT, 15 patients were without the abnormal gene, MRD were detected in 2 cases,and after decrement of immunosuppressive agents and donor lymphocyte infusion,MDR positive rate decreased to 1% and patients were still in complete remission,3 cases were with original genetic abnormality during 1 to 4 months after transplantation and relapsed origi nal diseases. Conclusion FISH detection can be simple,rapid,accurate and specific,and be great significant on the prognostic esti marion of allo-HSCT.