Rawstron A C, Owen R G, Davies F E, Johnson R J, Jones R A, Richards S J, Evans P A, Child J A, Smith G M, Jack A S, Morgan G J
Department of Haematology, The General Infirmary at Leeds.
Br J Haematol. 1997 Apr;97(1):46-55. doi: 10.1046/j.1365-2141.1997.72653.x.
The aim of this study was to develop a flow cytometric test to quantitate low levels of circulating myeloma plasma cells, and to determine the relationship of these cells with disease stage. Cells were characterized using five-parameter flow cytometric analysis with a panel of antibodies, and results were evaluated by comparison with fluorescent consensus-primer IgH-PCR. Bone marrow myeloma plasma cells, defined by high CD38 and Syndecan-1 expression, did not express CD10, 23, 30, 34 or 45RO, and demonstrated weak expression of CD37 and CD45. 65% of patients had CD19- 56+ plasma cells, 30% CD19- 56(low), and 5% CD19+ 56+, and these two antigens discriminated myeloma from normal plasma cells, which were all CD19+ 56(low). Peripheral blood myeloma plasma cells had the same composite phenotype, but expressed significantly lower levels of CD56 and Syndecan-1, and were detected in 75% (38/51) of patients at presentation, 92% (11/12) of patients in relapse, and 40% (4/10) of stem cell harvests. Circulating plasma cells were not detectable in patients in CR (n = 9) or normals (n = 10), at a sensitivity of up to 1 in 10,000 cells. There was good correlation between the flow cytometric test and IgH-PCR results: myeloma plasma cells were detectable by flow cytometry in all PCR positive samples, and samples with no detectable myeloma plasma cells were PCR negative. Absolute numbers decreased in patients responding to treatment, remained elevated in patients with refractory disease, and increased in patients undergoing relapse. We conclude that flow cytometry can provide an effective aternative to IgH-PCR that will allow quantitative assessment of low levels of residual disease.
本研究的目的是开发一种流式细胞术检测方法，以定量循环中低水平的骨髓瘤浆细胞，并确定这些细胞与疾病分期的关系。使用一组抗体通过五参数流式细胞术分析对细胞进行表征，并通过与荧光一致性引物IgH-PCR比较来评估结果。由高CD38和Syndecan-1表达定义的骨髓骨髓瘤浆细胞不表达CD10、23、30、34或45RO，并表现出CD37和CD45的弱表达。65%的患者有CD19 - 56+浆细胞，30%为CD19 - 56（低），5%为CD19+ 56+，这两种抗原可将骨髓瘤与正常浆细胞区分开来，正常浆细胞均为CD19+ 56（低）。外周血骨髓瘤浆细胞具有相同的复合表型，但CD56和Syndecan-1表达水平明显较低，在初诊患者中75%（38/51）可检测到，复发患者中92%（11/12）可检测到，干细胞采集样本中40%（4/10）可检测到。在CR患者（n = 9）或正常人群（n = 10）中未检测到循环浆细胞，检测灵敏度高达1/10000细胞。流式细胞术检测结果与IgH-PCR结果具有良好的相关性：在所有PCR阳性样本中，通过流式细胞术均可检测到骨髓瘤浆细胞，而未检测到骨髓瘤浆细胞的样本PCR为阴性。治疗有反应的患者绝对数量减少，难治性疾病患者数量仍居高不下，复发患者数量增加。我们得出结论，流式细胞术可为IgH-PCR提供一种有效的替代方法，用于定量评估低水平的残留疾病。
