三染色-流式细胞术检测急性B淋巴细胞白血病微量残留病的研究

Application of triple-color flow cytometry for minimal residual disease detection in acute B-lymphoblastic leukemia

目的 建立三染色 流式细胞术检测微量残留白血病细胞。方法 利用三染色 流式细胞仪检测急性B淋巴细胞白血病患儿骨髓中的CD1 0+ CD2 2+ CD45-或dim表现型白血病细胞。结果 在 2 3例急性B淋巴细胞白血病患儿骨髓中均可检测到CD1 0 + CD2 2 + CD45 -或dim 表现型细胞 ,检测敏感度约为10 - 4 。正常人骨髓、化疗后再生骨髓和长期完全缓解的白血病患儿的骨髓中未检测到CD1 0+ CD2 2+CD45-或dim 表现型细胞。该法可在化疗后 5 0d内监测到白血病细胞的变化 ,并且在骨髓细胞形态学确认复发之前即可检测到白血病细胞逐渐增加。结论 利用三染色 流式细胞仪检测骨髓中的CD1 0 +CD2 2+ CD45-或dim表现型白血病细胞可以敏感地监测到急性B淋巴细胞白血病患儿微量残留病的变化。

Objective To set up a sensitive triple color flow cytometric assay (FCM) for detection of residual leukemia cells. Methods Application of triple color flow cytometry to the patients for detecting CD 10 +CD 22 +CD 45 -or dim leukemic cells in acute B lymphoblastic leukemia (B ALL). Results Leukemic cells in B ALL patients' bone marrow (BM) consistently showed the CD 10 +CD 22 +CD 45 - or dim phenotype. FCM analysis could detect leukemic cells at the level of 1 per 10 4 cells. The CD 10 +CD 22 +CD 45 - or dim cell population was absent in normal BM, regenerating BM, and BM from leukemia patients in continuous complete remission. This assay could identify residual leukemic cells in 50 days after starting induction chemotherapy. Furthermore, a gradual increase of leukemic cell population could be monitored by this assay before morphological confirmation of BM relapse. Conclusion Triple color FCM was a sensitive method for detecting BM CD 10 +CD 22 +CD 45 - or dim leukemic cells and could be an additional useful tool to monitor MRD in B ALL patients.