Hematopathology Service, Department of Pathology and Laboratory Medicine, Memorial Sloan Kettering Cancer Center, New York, New York.
Curr Protoc. 2022 Aug;2(8):e503. doi: 10.1002/cpz1.503.
CLL/SLL is the most common leukemia in the western world. The disease is indolent; however, most patients require treatment at some point of the disease course. Outside of allogeneic transplants, the treatment is rarely curative but often controls CLL/SLL manifestations for many years. Several lines of therapy may be used sequentially to prolong clinical remission. Because of the prolonged disease course, CLL/SLL monitoring represents a sizable portion of the workload in a typical flow cytometry laboratory involved in the diagnosis and monitoring of hematopoietic neoplasms. Minimal/measurable disease monitoring of CLL/SLL has emerged as a key component in treatment monitoring and sequencing. In the face of effective therapies, clinical laboratories are tasked with monitoring ever smaller proportions of MRD with high precision and accuracy. With the recent addition of surface antigen-targeting biologics such as antibodies and CAR-T cells, the task has become more complex due to the unavailability of commonly analyzed antigens for flow cytometric analysis. This article details a flow cytometric test developed at Memorial Sloan Kettering Cancer Center that has proven to consistently achieve high sensitivity (<0.01% of nucleated cells) in the bone marrow and peripheral blood, even when CD19 is lost or unavailable for analysis. Moreover, the test helps distinguish between CLL and other CD5-positive B cell neoplasms. The Basic Protocol provides a detailed operational procedure for processing, staining, and cytometric acquisition of samples. The Support Protocol provides typical steps and caveats for MRD data analysis in CLL/SLL and in distinguishing CLL/SLL from other B cell neoplasms and normal CD5-positive B cells. © 2022 Wiley Periodicals LLC. Basic Protocol: Processing, staining, and cytometric analysis of bone marrow or peripheral blood cells for MRD analysis of CLL/SLL Support Protocol: Analysis and interpretation of CLL MRD assay.
CLL/SLL 是西方世界最常见的白血病。该疾病进展缓慢；然而，大多数患者在疾病过程的某个阶段需要治疗。除了异基因移植外，该治疗很少能治愈，但通常可以控制 CLL/SLL 多年的表现。几种治疗方案可能会依次使用以延长临床缓解期。由于疾病过程较长，CLL/SLL 监测在涉及造血肿瘤诊断和监测的典型流式细胞术实验室的工作量中占很大一部分。CLL/SLL 的最小/可测量疾病监测已成为治疗监测和排序的关键组成部分。在有效治疗的情况下，临床实验室的任务是监测越来越小比例的 MRD，同时保持高精度和准确性。随着最近表面抗原靶向生物制剂（如抗体和 CAR-T 细胞）的加入，由于流式细胞术分析中无法获得常用的分析抗原，任务变得更加复杂。本文详细介绍了 Memorial Sloan Kettering 癌症中心开发的一种流式细胞术测试，该测试已被证明在骨髓和外周血中始终具有很高的灵敏度（<0.01%有核细胞），即使 CD19 丢失或无法进行分析时也是如此。此外，该测试有助于区分 CLL 和其他 CD5 阳性 B 细胞肿瘤。基本方案提供了处理、染色和样本流式细胞术采集的详细操作程序。支持方案提供了 CLL/SLL 中 MRD 数据分析以及区分 CLL/SLL 与其他 B 细胞肿瘤和正常 CD5 阳性 B 细胞的典型步骤和注意事项。©2022Wiley Periodicals LLC.基本方案：CLL/SLL 微小残留病分析中骨髓或外周血细胞的处理、染色和流式细胞术分析支持方案：CLL MRD 检测分析和解释。
