骨髓增生异常综合征del（20q）异常细胞克隆在骨髓细胞系列和凋亡细胞中的分布

为了探讨20号染色体长臂部分缺失del（20q）]的1例骨髓增生异常综合征（MDS—RAEB-1）患者异常细胞克隆在骨髓各细胞系列和凋亡细胞中的分布，应用单克隆抗体通过流式细胞分选（FACS）del（20q）的MDS患者骨髓CD15^＋、GPA^＋、CD3^＋CD56^-CD16^-、CD19^＋、CD3^-CD56^＋CD16^＋细胞，应用Annexin V—FITC和PI通过FACS分选该患者骨髓Annexin V^＋PI^-（凋亡细胞）和Annexin V^-PI^-细胞（活细胞），然后应用LSID20S108探针和Telvysion^TM 20p探针对分选细胞分别进行间期双色荧光原位杂交（D—FISH）检测。同时选取4例核型正常者作为对照组进行相应检测。结果显示。该MDSRAEB-1患者的骨髓CD15^＋细胞和GPA^＋细胞中MDS克隆百分比分别为70．50％和93．33％，明显高于对照组（5．39％和6．17％）；而在CD3^＋CD56^-1CD16^-1，CD19^＋，CD3^-CD56^＋CD16^＋细胞中分别为3．23％、4．32％、5．77％，低于对照组（5．76％、4．85％、6．36％）。该患者骨髓有核细胞凋亡率为16．09％。MDS细胞克隆在凋亡细胞和活细胞中的比例分别为32．48％和70．11％。结论：MDS患者del（20q）异常克隆主要分布于粒系与红系细胞，以及一小部分凋亡细胞

Distribution of Abnormal Cell Clone with Deletion of Chromosome 20q in Marrow Cell Lineages and Apoptosis Cells in Myelodysplastic Syndrome

This study was aimed to investigate the distribution of abnormal clone in marrow cell lineages and apoptosis cells in myelodysplastic syndrome (MDS) with deletion of chromosome 20q. Monoclonal antibodies recognizing myeloid precursors (CD15), erythroid precursors (GPA), T cells (CD3(+)CD56(-)CD16(-)), B cells (CD19), NK cells (CD3(-)CD56(+)CD16(+)) were used to sort bone marrow cells in a MDS patient with del (20q) by fluorescence activated cell sorting (FACS). Annexin V-FITC and PI were used to sort bone marrow Annexin V(+)PI(-) and Annexin V(-)PI(-) cells by FACS. The sorted positive cells were detected by interphase dual-color fluorescence in situ hybridization (D-FISH) using a LSI D20S108 probe (Spectrum Orange) and a Telvysion TM 20p probe (Spectrum Green). FACS and FISH analysis were also performed on the samples from 4 cases with normal karyotype. The results showed that the proportions of MDS clone in the myeloid and erythroid precursors were 70.50% and 93.33% respectively, in the RAEB-1 patient with del (20q) and were obviously higher than that in control group (5.39% and 6.17%). The proportions of abnormal clone in T, B and NK cells were 3.23%, 4.32% and 5.77% respectively and were less than that in control group (5.76%, 4.85%, 6.36%). The percentage of apoptotic cells in the bone marrow nucleated cells was 16.09%. The proportions of MDS clone in Annexin V(+)PI(-) and Annexin V(-)PI(-) cells were 32.48% and 70.11%, respectively. It is concluded that most myeloid and erythroid precursors are originated from the abnormal clone in MDS with del (20q). A little part of apoptotic cells are derived from the abnormal clone.