Quantification of adult T-cell leukemia/lymphoma cells using simple four-color flow cytometry
Quantification of adult T-cell leukemia/lymphoma cells using simple four-color flow cytometry
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DOI：
10.1515/cclm-2014-0183
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发表时间：
2015-01-01
影响因子：
6.8
通讯作者：
Watanabe, Nobukazu
中科院分区：
文献类型：
--
作者：
Ishigaki, Tomohiro;Zaike, Yuji;Watanabe, Nobukazu
背景资料：外周血中成人T细胞白血病/淋巴瘤（ATL）细胞的绝对数量是评估疾病状态的重要指标。然而，显微镜下计数ATL细胞的形态学的基础上需要的经验，往往是不准确的，由于ATL.Methods的稀有性：基于我们的研究表明，急性型ATL细胞是专门富集在CD 4 +/CD 7-（CD 7 N）的馏分，一种新的分析方法，以准确地量化ATL细胞建立使用内部珠标准和简单的四色流式细胞术。结果：流式细胞术检测的CD 7 N细胞数与有经验的技术人员在显微镜下检测的异常淋巴细胞数有很强的相关性[Pearson 'sR，0.963;斯皮尔曼rho，0.921;互相关系数0.962]。线性回归系数接近1（β =1.013）。我们的方法可以检测1个细胞/μ L，定量限为2.9和9.8个细胞/μ L之间。CD 4+细胞中CD 7 N细胞的比例在化疗期间发生变化，反映了化疗敏感和耐药病例之间的差异。Kaplan-Meier分析与对数秩检验表明，化疗后CD 7 N比例下降的患者有显着较长的疾病特异性生存期（p=0.003）。结论：我们新建立的方法定量肿瘤细胞急性型ATL患者。该方法可用于评价化疗的疗效，而CD 7 N比例的变化对判断预后有重要意义。
Background: The absolute number of adult T-cell leukemia/lymphoma (ATL) cells in peripheral blood is an essential indicator to evaluate disease status. However, microscopically counting ATL cells based on morphology requires experience and tends to be inaccurate due to the rarity of ATL.Methods: Based on our research showing that acute-type ATL cells are specifically enriched in the CD4+/CD7- (CD7N) fraction, a new analytical method to accurately quantify ATL cells was established using an internal bead standard and simple four-color flow cytometry. This method was verified by comparison with microscopic examination of 49 peripheral blood samples and used to follow up patients.Results: A strong correlation was observed between the number of CD7N cells measured by flow cytometry and the number of abnormal lymphocytes measured microscopically by experienced technicians [Pearson's R, 0.963; Spearman's rho, 0.921; intercorrelation coefficient, 0.962]. The linear regression coefficient was close to 1 (beta=1.013). Our method could detect 1 cell/mu L, and the limit of quantitation was between 2.9 and 9.8 cells/mu L. The frequency of CD7N cells among CD4+ cells changed during chemotherapy, which reflected differences between chemosensitive and chemoresistant cases. Kaplan-Meier analysis with a log-rank test showed that patients with decreased CD7N proportion after chemotherapy had significantly longer disease-specific survival (p=0.003).Conclusions: Our newly established method quantified tumor cells in patients with acute-type ATL. Furthermore, this method was useful for assessing the efficacy of chemo-therapy, and the change of the CD7N proportion could be more important to predict prognosis.