应用流式细胞术分析骨髓增生异常综合征患者原始细胞免疫表型及叶酸受体的表达

Flow cytometry analysis of immunophenotype features and folate receptor expression of blasts in myelodysplastic syndromes

目的研究骨髓增生异常综合征（MDS）患者骨髓原始细胞免疫表型及叶酸受体（FR）的表达。方法应用四色流式细胞术对常规及二次设门选取的MDS患者骨髓原始细胞、CD34^＋原始细胞进行免疫表型分析及叶酸受体的表达检测，并以急性髓系白血病M2型（AML-M2）作阳性对照。结果随着MDS疾病的进展（RA／RAS→RAEB→RAEB-T），应用常规设门选取的原始细胞中CD矗原始细胞比例逐渐升高，各亚型间差异有统计学意义（P〈0．05）；HLA-DR及髓系抗原CD117、CD13、CD33表达逐渐增高，CD15，表达逐渐降低，差异有统计学意义（P〈0．05）。而应用二次设门取CD矗原始细胞行抗原表达分析，同样异常表达HLA_DR及髓系抗原CD117、CD13、CD33且表达率高于常规分析法（P〈0．05），但各相关抗原表达率在各亚型间差异无统计学意义（P〉0．05）。用常规方法直接提取的MDS各亚型的原始细胞及二次设门提取的CD玉原始细胞上均未得到FR表达阳性的细胞。结论随着MDS疾病的进展，原始细胞表面抗原向髓系更加不成熟的表型转变，但髓系抗原的异常表达只反映了优势恶性克隆数量上的增加而不能真正反映疾病进展的本质。单纯应用流式细胞技术尚不能检测出MDS原始细胞上是否存在叶酸受体的表达。

Objective To explore the immunophenotype features and folate receptor expression of blasts in patients with myelodysplastic syndromes（MDS）. Methods Four-color flow cytometry using conventional and secondary gating strategies was used into analysis the immunophenotype features and folate receptor（FR） expression of blasts and CD34^＋ cells in bone marrow nucleated cells with MDS. The patients with acute myeloid leukemia-M2 （AML-M2） were as positive control. Results With progression of MDS from RA/RAS, RAEB to RAEB-T, using conventional gating strategy, the proportion of CD34^＋ cells were gradually increased（P 〈 0.05 ）. Moreover, the expression of HLA-DR, CD117, CD13, CD33 were also gradually increased and the expression of CD15 was gradually decreased （ P 〈 0.05）. Using secondary gating strategy, the expression of HLA-DR, CD117, CD13, CD33 on blasts were higher than those by conventional gating（ P 〈 0.05）. However, there was no significant difference（P〉 0.05） in the expression of above mentioned antigens on CD34^＋ cells among different MDS subtypes. On the other hand, there were no expression of FR on blasts and CD34^＋ cells with different MDS subtypes. Conclusion With progression of M DS, the antigens of blasts surface change into more immature immunophenotype of medullary system. But these antigens abnormal expression only illustrates the increase of ascendant malignant clone quantity, it can not reflect the nature of the disease. Using flow cytometry technique can not detect whether or not FR expression on the blasts with MDS.