四色流式细胞术分析骨髓增生异常综合征原始细胞免疫表型

为了研究骨髓增生异常综合征（MDS）患者骨髓原始细胞免疫表型特征，应用四色流式细胞术对常规设门及二次设门策略选取的29例MDS患者的骨髓细胞悬液中，CD34^＋细胞进行免疫表型分析。结果表明：①随着MDS的进展（RA／RAS→RAEB→RAEB—T），根据CD45／SSC图中选取的原始细胞群中，CD34^＋细胞比例逐渐增高．分别为8．0％、46．4％、76．8％，各亚型之间差别有显著意义（P〈0．05）。②在CD45 vs SSC散点图上，设门所取的原始细胞抗原表达随着RA／RAS→RAEB→RAEBT的进展，HLA—DR、CD13、CD33、CD117表达逐渐增高，CD15表达逐渐减低，差别均有显著意义（P〈0.05）。③在CD45VSCD34散点图上二次设门取CD34^＋原始细胞行抗原表达的分析，同样异常表达HLA—DR及髓系抗原CD13、CD33和CD117，比例高于常规分析法（P〈0．05），CD15比例减低，在RA／RAS中CD13、CD33、CD117和HLA—DR的表达较常规分析的原始细胞比例明显增高（P〈0.05），相关抗原的表达率在各亚型间差别无显著意义（P〉0．05）。结论：常规设门法能反映MDS疾病的进展，而二次设门策略则更能反映MDS原始细胞的生物学特征，CD34的异常表达代表原始细胞的不成熟性和异质性，在临床高度怀疑但形态学和细胞遗传学不能诊断时，原始细胞免疫分型对MDS的诊断更具意义。

Four-color Flow Cytometric Immunophenotypic Features of Blasts in Myelodysplastic Syndromes

This study was aimed at exploring the immunophenotypic features of blasts in patients with myelodysplastic syndromes (MDS). Four-color flow cytometry using conventional and secondary gating strategies was used to immunophenotype blasts and the CD34 positive cells in bone marrow nucleated cells of 29 patients with MDS. The results showed: (1) with progression of MDS from RA/RAS, RAEB to RAEB-T, the proportion of CD34~+ cells were gradually increased from 8.0%, 46.4% to 76.8% (P<0.05); (2) using CD45 vs SSC gating strategy, with the transformation of RA/RAS, RAEB to RAEB-T, the expression of HLA-DR, CD13, CD33, CD117 were also gradually increased (P<0.05), and the expression of CD15 was gradually decreased (P<0.05); (3) using CD45 vs CD34 gating strategy, the expression of HLA-DR, CD13, CD33, CD117 on blasts were higher by secondary gating method than those by conventional gating (P<0.05). However, there were no significant difference (P>0.05) in the expression of above-mentioned antigens on CD34~+ cells among different MDS subtypes. It is concluded that conventional gating method can reflect MDS progression from RA/RAS, RAEB to RAEB-T, and secondary gating strategy may accurately reflect the biological features of blasts in MDS. Abnormal expression of CD34 is related to the immaturity level and heterogeneity of blast cells, which is beneficial to the diagnosis of clinically suspected MDS incapable of diagnosing with morphology and cytogenetics.