BIOLOGICAL STUDIES OF PCR POSITIVITY IN CML

接受干扰素或骨髓移植治疗的CML患者可以进入 长期的细胞遗传学缓解。令人惊讶的是，其中只有一部分 患者最终会复发。因为慢性粒细胞白血病患者患有癌症- 特异性标记--导致bcr-abl基因异常的t(9；22)和 蛋白质--检测到极低水平的微小残留病是 现在，随着FISH和基于PCR的技术的使用，这一点成为可能。然而， 微小残留病的临床相关性尚不清楚，因为 这些患者预后的异质性。例如，骨髓 移植受者和接受干扰素治疗的患者可能会表现出 残留的bcr-abl阳性细胞或细胞遗传学阳性前夕 临床复发。相反，我们也观察到了一例PCR阴性的慢性粒细胞白血病。 患者在几个月内复发。因此，我们建议 对慢性粒细胞白血病患者细胞遗传学缓解情况进行系统分析 移植、干扰素或化疗方案治疗后 (在本P01中概述)以确定(I)是否有低水平的残留病 通过聚合酶链式反应或FISH在骨髓中检测到 Bcr-abl信息在能够分裂和形成克隆的细胞中的表达；(Ii) 如果对PCR阳性菌落百分比的研究增加了与 PCR检测可以帮助确定哪些患者的残留最小 疾病会复发；以及(Iii)如果bcr-abl的差异表达 MESSAGE和p210/bcr-abl在不同时期造血细胞中的表达 成熟(通过细胞分离技术和特定阶段获得 菌落分析)的存在。这些实验将在不同的环境中进行 PCR阳性的患者类别，如同种异体移植后的患者 或者鱼呈阳性和复发，而不是那些不复发的人。我们的 结果将与实验结果进行比较。 间期VS中期FISH，试图了解临床 微小残留疾病患者的不同亚组和TO 阐明少量Ph+细胞保留的机制 没有复发的发生。

CML patients treated with interferon or bone marrow transplant can enter long-term cytogenetic remissions. Surprisingly, only a portion of these patients will eventually relapse. Because CML patients carry a cancer- specific marker--the t(9;22) resulting in an aberrant BCR-ABL gene and protein--the detection of very low levels of minimal residual disease is now possible with the use of FISH and PCR-based technologies. However, the clinical relevance of minimal residual disease is unclear because of the heterogeneity of outcome in these patients. For instance, bone marrow transplant recipients and interferon-treated patients may show evidence of residual BCR-ABL- positive cells or eve of cytogenetic positivity without clinical relapse. Conversely, we have also observed a PCR-negative CML patient who relapsed within several months. We therefore propose to undertake a systematic analysis of CML patients in cytogenetic remission after treatment with the transplant, interferon, or chemotherapy programs (outlined in this P01) to determine (i) if low levels of residual disease detected by PCR or FISH in bone marrow are accounted for by the presence of BCR-ABL message in cells capable of dividing and forming colonies; (ii) if the study of the percentage of PCR-positive colonies adds relevance to the PCR assay and can help determine which patients with minimal residual disease will relapse; and (iii) if differential expression of the BCR-ABL message and p210/BCR-ABL in hematopoietic cells at different stages of maturation (obtained by cell separation techniques and by stage-specific colony assays) exists. These experiments will be performed in various categories of patients such as those after allograft who are PCR-positive or FISH-positive and relapse as opposed to those who do not relapse. Our results will be compared to those obtained from experiments with interphase vs metaphase FISH in an attempt to understand the clinically different subsets of patients with minimal residual disease and to elucidate the mechanisms by which small numbers of Ph+ cells remain without the occurrence of relapse.