應用流式細胞儀建立正常骨髓紅血球分化圖形

流式細胞儀發展至今已被廣泛使用於白血病的細胞標記檢測。本科六年前與美國西雅圖血液邏輯診斷實驗室（Hematologics Inc.）共同合作，係利用細胞標記套組做為白血病診斷、預後與治療追蹤之用。根據文獻，當出現不同分化系列之細胞標記、同分化系列卻表現不同時期之細胞標記、細胞標記表現量超過或低於正常量、細胞數量與比例分佈不正確皆可視為異常表現。再者，骨髓再生不良等紅血球分化異常疾病，紅血球系列可能有CD71表現下降、早期細胞表現CD117等現象。目前除形態學方法外，尚未有既定標準判別phenotype異常方法。本研究係挑選無貧血症狀之白血病患者常規骨髓，同時利用三種螢光紅血球細胞標記（CD235a-FITC、CD71-PE、CD45-perCP）以流式細胞儀進行檢測後，並且嘗試最佳化儀器設定。以CD71（Y軸）及CD235a（X軸），建立四區正常骨髓紅血球分化圖形：第一區為主要為Erythroblsat及Basophilic Normoblast，CD71^(dim/bright)（螢光亮度10^2～10^3）／CD235a^-；第二區主要為Polychromatophilic Normoblast及Orthochromatophilic Normoblast，佔四區比例最高，CD71^+（螢光亮度10^3～10^4）／CD235a^+（螢光亮度10^2～10^3）；第三區與第四區為主要為Reticulocyte及成熟RBC，CD71^-/CD235a^+（螢光亮度10^2～10^3）。當建立正常紅血球分化圖形後，之後便可以上述規則比對紅血球分化異常病患各區細胞分布比例與細胞標記亮度表現來幫助臨床診斷。

關鍵字

流式細胞儀；紅血球分化；細胞標記

並列摘要

Detection of leukemia cells can be achieved by Flow cytometry which is also developed for diagnosis and prognosis of diseases including minimal residual disease. In collaboration with the American Hematologicsinc., we have been studying on distinction of the abnormal blast cells from normal precursor cells by the CD marker panels, and flow cytometry. Previous study has indicated the expression of abnormal CD markers is closely related to lineage infidelity, maturational asynchrony, antigenic absence and quantitative abnormality. Furthermore, in Myelodysplastic syndrome and other erythroid differentiation disorders, the expression of CD71 in erythroid cells may be reduced and, and that of CD117 may be increased in early cell expression. In this study, using flow cytometry, the clinical routine samples from bone marrows of leukemia patients without anemia were monitored and statistically analyzed. The CD markers analyzed included CD235a-FITC, CD71-PE and CD45-perCP in Erythropoiesis. After optimization, results using CD71 as Y-axis and the CD235a as X-axis, which forma the four districts, were used to establish normal bone marrow erythroid differentiation pattern: The first district is mainly erythroblsat and the basophilic normoblast of CD71^(dim/bright) (fluorescence intensity 10^2~10^3)/CD235a^-. The second district comprises mostly the polychromatophilic normoblast, and the orthochromatophilic normoblast, which occupies the highest cell proportion of CD71^+ (fluorescence intensity10^3-10^4)/CD235a^+ (fluorescence intensity 10^2-10^3) in four districts. The fourth district was mainly mature Reticulocyte with the CD71^-/CD235a^+, (fluorescence intensity 10^2-10^3). Our results indicated the flow cytometry in combination with CD markers can be used to help clinical diagnosis of erythroid differentiation in abnormal patients.

並列關鍵字

Flow cytometry ； Erythropoiesis ； CD markers

國際替代計量

應用流式細胞儀建立正常骨髓紅血球分化圖形

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