ABSTRACT
ObjectiveTo investigate the current situation of immunophenotyping of acute leukemia by flow cytometry in China (except Taiwan, Hongkong and Macau), and to provide theoretical evidence for the formulation of guidelines of immunophenotyping of leukemia by flow cytometry.MethodsThe Chinese Hospital Knowledge Database(CHKD) was used to retrieve the1994-2009 published papers on immunophenotyping of acute leukemia by flow cytometry.All of 380 retrieved papers were analyzed by the GraphPad Prism 4. ResultsThe number of published papers in China increased yearly and changed gradually from large cities to small and medium-sized cities in local distribution. The most often reported area was Beijing, followed by Jiangsu and Guangdong. In specimen processing, the ratios of using mononuclear cell labling method and whole blood labling-lysing-washing method were 34.2 ％ (113/330) and 65.8 ％ (217/330) respectively. The ratios of using single color, two color, three color and four color labling method were 15.4 ％ (53/344), 13.7 ％ (47/344), 54.1％ (186/344) and 16.3 ％ (56/344), respectively. The number of fluorescent antibodies was used from less than 10 to more than 20.Most of them were between 11 to 16,which was 48.6 ％ (167/344) of total number of published papers. In data acquisition and analysis, the ratios of using FSC/SSC gating and CD45/SSC gating were 34.2 ％ (113/330) and 65.8 ％ (217/330), respectively. There were some differences in the positive criteria of lymphatic and myeloid and cytoplasm antigens.The main positive criteria of lymphatic, myeloid antigen was ≥20 ％, which was 64.8 ％ (223/344) and 95.4 ％ (328/344) . The main positive criteria of cytoplasm antigen was ≥ 10 ％, which accounted for 61.2 ％ (156/255). Conclusion A common view has been reached in specimen processing,gating method,data acquisition and analysis in immunophenotyping of acute leukemia by flow cytometry. But there were still some difference in the number of fluorescent antibodies, panels and evaluation of positive criteria of antigens.
ABSTRACT
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 CD+34 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 CD+34 cells were gradually increased(P＜0.05).Moreover,the expression of HLA-DR,CDll7,CD13,CD33 were also gradually increased and the expression of CDl5 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 CD+34 cells among different MDS subtypes.On the other hand,there were no expression of FR on blasts and cD+34 cells with different MDS subtypes.Conclusion With progression of MDS,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.