Babusíková O, Glasová M, Stasáková J, Kusenda J, Koníková E
Cancer Research Institute, Slovak Academy of Sciences, Bratislava.
Neoplasma. 1997;44(6):348-55.
In our study we used for definition of leukemia/lymphoma cells a new parameter which allows the enumeration of mean fluorescence intensity expressed by the number of antigen molecules per cell. Quantitative immunofluorescence using calibration microbeads was performed in 36 patients with different acute and chronic lymphoid and myeloid leukemia and in 19 healthy volunteers. We showed that quantitative immunophenotyping allowed the definition of aberrant marker densities on neoplastic cells. We demonstrated under- and overexpression of CD8 marker in CD3/CD4/CD8 complex in T acute lymphatic leukemia and T non-Hodgkin's lymphoma and T leukemia of large granular lymphocytes as compared to normal counterparts. We pointed out that certain antigens (e. g. CD10, CD4, CD24) were expressed at different levels on different cell subsets (CD10 in early B-acute lymphatic leukemia and coexpressed in T-acute lymphatic leukemia, CD4 on T cells and monocytes, CD24 on B cells and granulocytes in chronic myeloid leukemia). We showed that quantitative immune fluorescence could provide new data contributing to a more precise definition of cell differentiation. We documented the significant difference between antigen density of early and late markers in B-cell and myeloid malignancies. Further, we demonstrated that quantitative immune phenotyping could help in determination of exact definition of pathologic clone in morphologically immature leukemia population and showed that parameters of this method are also convenient for cytoplasmic marker evaluation. In our study we were able to demonstrate that CD45 quantitative expression appeared to be a more informative parameter than its percentage of antigen-positive cells as a measure of antigen expression only and we pointed out that low and high CD45 densities enabled to differentiate between pathological clone and residual healthy population in examined sample. We showed that quantitative immune phenotyping could be another important parameter for definition of leukemia phenotype suitable for detection of minimal residual disease.
在我们的研究中，我们使用了一个新参数来定义白血病/淋巴瘤细胞，该参数能够通过每个细胞上抗原分子的数量来计算平均荧光强度。我们对36例不同类型的急性和慢性淋巴细胞及髓细胞白血病患者以及19名健康志愿者进行了使用校准微珠的定量免疫荧光检测。我们发现定量免疫表型分析能够确定肿瘤细胞上异常标志物的密度。与正常对照相比，我们证实了在T急性淋巴细胞白血病、T非霍奇金淋巴瘤以及大颗粒淋巴细胞性T白血病中，CD3/CD4/CD8复合物中CD8标志物存在表达不足和过度表达的情况。我们指出某些抗原（如CD10、CD4、CD24）在不同细胞亚群上的表达水平不同（早期B急性淋巴细胞白血病中的CD10以及T急性淋巴细胞白血病中共表达的CD10；T细胞和单核细胞上的CD4；慢性髓细胞白血病中B细胞和粒细胞上的CD24）。我们表明定量免疫荧光能够提供有助于更精确地定义细胞分化的新数据。我们记录了B细胞和髓系恶性肿瘤中早期和晚期标志物抗原密度之间的显著差异。此外，我们证明定量免疫表型分析有助于确定形态学上不成熟的白血病群体中病理克隆的确切定义，并且表明该方法的参数也便于评估细胞质标志物。在我们的研究中，我们能够证明CD45的定量表达似乎比仅作为抗原表达量度指标的抗原阳性细胞百分比更具信息性，并且我们指出低和高CD45密度能够区分检测样本中的病理克隆和残余健康群体。我们表明定量免疫表型分析可能是定义白血病表型的另一个重要参数，适用于检测微小残留病。
