Tousková M, Maisnar V, Krejsek J, Kopecký O
Ustav klinické imunologie a alergologie Fakultní nemocnice LF UK, Hradec Králové.
Vnitr Lek. 1999 Dec;45(12):708-12.
The advent of new therapeutic approaches to multiple myeloma made necessary the introduction of novel methods for detection of minimal residual disease. Among others approaches residual disease can be detected by the immunofluorescence using flow cytometry. We have examined the co-expression of CD19, CD38, CD45, CD54, CD56, and CD138 molecules in cells of peripheral blood and bone marrow aspirates in patients with multiple myeloma by 3-color flow cytometry. For the detection and characterization of multiple myeloma cells, combinations of following antibodies were used: anti-CD19 FITC, anti-CD38 FITC, anti-CD38 PE, anti-CD54 FITC, anti-CD56 PE-Cy5, anti-CD45 PE, anti-CD45 PE-Cy5 (Immunotech) and anti CD138 PE (Serotec). The samples were analyzed using EPICS XL (Coulter) flow cytometer, and the analysis was based on at least 10,000 events. Samples from 17 patients were analyzed. The percentage of multiple myeloma cells ranged between 0.3% and 54.2% in bone marrow aspirates and between 0.0 and 11.8% in periferal blood. The expression of CD138, CD38, CD54 and CD56 molecules was found in 100%, 100%, 85% and 68% of examined cases, respectively. In our opinion, multiple myeloma cells are best characterized by following combinations of antibodies: CD38 FITC/CD138 PE/CD45 PE-Cy5, CD54 FITC/CD138 PE/CD56 PE-Cy5 or CD54 FITC/CD38 PE/CD 56 PE-Cy5. The identification of a malignant clone is the first and the most important step in the characterization of the disease, determination of its prognosis and the detection of residual disease after treatment. Three-color flow cytometry represents a method which can meet these goals.
针对多发性骨髓瘤的新治疗方法的出现，使得引入检测微小残留病的新方法成为必要。在其他方法中，残留病可通过流式细胞术的免疫荧光法检测。我们通过三色流式细胞术检测了多发性骨髓瘤患者外周血和骨髓穿刺液细胞中CD19、CD38、CD45、CD54、CD56和CD138分子的共表达情况。为了检测和鉴定多发性骨髓瘤细胞，使用了以下抗体组合：抗CD19 FITC、抗CD38 FITC、抗CD38 PE、抗CD54 FITC、抗CD56 PE-Cy5、抗CD45 PE、抗CD45 PE-Cy5（Immunotech）和抗CD138 PE（Serotec）。使用EPICS XL（库尔特）流式细胞仪对样本进行分析，分析基于至少10000个事件。对17例患者的样本进行了分析。骨髓穿刺液中多发性骨髓瘤细胞的百分比在0.3%至54.2%之间，外周血中在0.0%至11.8%之间。在所检测的病例中，分别有100%、100%、85%和68%的病例发现了CD138、CD38、CD54和CD56分子的表达。我们认为，多发性骨髓瘤细胞最适合用以下抗体组合来鉴定：CD38 FITC/CD138 PE/CD45 PE-Cy5、CD54 FITC/CD138 PE/CD56 PE-Cy5或CD54 FITC/CD38 PE/CD 56 PE-Cy5。识别恶性克隆是疾病特征化、确定其预后以及治疗后检测残留病的首要且最重要的步骤。三色流式细胞术是一种能够实现这些目标的方法。
