IMMUNOPHENOTYPE ANALYSE OF ACUTE LEUKEMIA BY FLOW CYTOMETRY WITH CD_(45) GATING
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Objective:To study the immunophenotype of acute leukemia and its clinical significance.Methods:A panel of lineageassociated monoclonal antibodies,CD 45 and direct immunofluorescence technique were used to analyse the immunophenotype in 34 cases of acute leukemia.Results:There were 8 B ALLs and 3 T Alls in 11 cases of ALL,in which of them 4 cases were expressed with myeloid antigens(36.4%),and 10 cases were expressed with CD 34 (90.1%).Meanwhile there were 7 cases with lymphoid antigens expression(29.17%),16 cases with CD 34 and DR expression (70.8%) in 24 cases of AML,and no CD 34 and DR antigen expression were found in 5 M 3 patients.Furthermore,CR rate in myeloid antigen positive ALL is slight lower than in that myeloid antigen negative ALL(1/3 vs 5/6),but there was no significant difference between them( P 0.05).CR rate in lymphoid antigen positive AML is markly lower than in that lymphoid antigen negative AML(0/5 vs 10/10, P 0.01).Conclusion:It was the best method by flow cytometry on CD 45 versus sidescatter gating for immunophenotype analysis of luekemia,and aberrant antigen expression in leukemia was recognized to be a fact of poorer prognosis.Keywords:
Immunophenotyping
Immunofluorescence
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Immunophenotyping detected by flow cytometry and its clinical significance in acute myeloid leukemia
Objective To investigate the immunophenotypic characteristics of acute myeloid leukemia (AML)and its relationship with chemotherapy response.Methods The immunophenotyping was performrd by flow cytometry in 96 newly diagnosed AML patients.Results In 96 patients with AML,the myeloid antigens were mainly expressed including CD13(94/96,97.92％),CD117(86/96,89.58％),CD33(79/96,82.29％)respectively.The positive expression of stem/progenitor cell differentiation antigens were CD38 (92/96,95.83％),HLA-DR(72/96,75.00％)and CD34(60/96,62.50％).While CD34 and HLA-DR were fewly expressed in M3.The CD14 was only expressed in M4 and M5.The lymphocytic antigens had positive expression in the patients with AML,in which CD4 was 37.5％(36/96),CD7 was 22.92％(22/96)and CD2 was 10.42 ％(10/96).In 46 patients for treatment response analysis,total CR rate was 67.39％(31 /46).There was no significant difference of CR rates between the antigens positive group and negative group including CD33,CD117,CD11b,CD14,CD64,CD4,CD34 and HLA-DR.However,the CR rate was significantly lower in CD7 positive cases than CD7 negative ones,with statistically significance difference(P=0.01).Conclusions CD13,CD33and CD117 are the most common antigens in AML.The patients with the positive expression of CD7 have poor response to chemotherapy in AML.
Key words:
Leukemia,myeloid,acute; Immunophenotyping; Flow cytometry
Immunophenotyping
CD117
CD64
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Objective:To explore the characteristic of immunophenotyping in acute myeloid leukemia(AML).Methods:A panel of monoclonal antibodies (MoAb) and indirect immunofluorescence assay by fluoromicroscope was used to determine the pretherapy immunophenotype of 70 AML patients.Results:Cells from all cases were labeled by at least one of the anti myeloid McAb.The order of myeloid markers expression in AML was as fallows:CD 33 CD 13 CD 15 ,CD 9 was positive,CD 34 and HLA DR were negative in all M 3 patients.The blast cells of 16 patients(30%) expressed antigen CD 34 .The patients with CD 34 positive were no significantly different from the remaining 54 patients with CD 34 negative with respect to age,blood counts and the proportion of blast cells in blood and bone marrow,but their blasts more likely to express HLA DR,CD 38 ,CD 7 and lack the CD 15 .The expression of lymphoid antigens in 70 cases of AML observed at our institution,16 cases(23%) carried lymphoid antigens.CD + 4 6 cases(13.8%),incidence of CD 4 expression in each FAB subtype was as follows:M 2 8.8%,M 4 60%;CD + 7 9 cases(16.9%),M 1 50%,M 2 18%,M 5b 16.7%.The typical characteristic in CD + 4 AML was CD 34 low CD 34 high.Conclusion:Combination between the expression of CD 9 and other markers can provide basis for clinical diagnosis.CD + 34 AML. cases often had FAB M 1 morphologic characteristics and lower complete remission(CR) rate.This result demonstrated that CD 34 positive AML is poorly differentiated.These data collectively indicate that CD + 7 and CD + 4 AML patients showed a poorer prognosis.The cellular and clinical characteristics of CD + 7 are recognized to be a distinct category of AML.
Immunophenotyping
Immunofluorescence
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This study was purposed to investigate the immunophenotype characteristics and their significance in subtypes of acute lymphoblastic leukemia (ALL). The immunophenotypes in 40 cases of ALL were analyzed with three color flow cytometry using CD45/SSC two-parametric gating. The results showed that the three color flow cytometry assay using CD45/SSC two-parametric gating could accurately distinguish each other from lymphocytes, monocytes, granulocytes, erythroblasts and primitive cells in bone marrow and/or peripheral blood. Among 40 cases of ALL, B-ALL was 26 cases, T-ALL was 11 cases, HAL was 3 cases. All of the 26 cases of B-ALL expressed CD19 with positive rate of 100%, meanwhile 11 cases of T-ALL most highly expressed CD17 with positive rate of 100%. 12 cases of ALL with myeloid antigen expression (My-ALL) were involved in ALL, the incidence of these cases was almost 30% (12/40). The CD13 was expressed most highly in myeloid antigens. All 3 cases of HAL coexpressed myeloid and B-lineage antigens, among them CD34 was expressed in 2 cases with positive rate of 66.67%. It is concluded that three color flow cytometry assay using CD45/SSC two-parametric gating can exclude the interference of normal cells, thereby the results are more reliable and more accurate.
Immunophenotyping
Cytometry
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Immunophenotyping has become common in the diagnosis and classification of leukemia. To evaluate the immunophenotype of acute myeloid leukemia (AML), multiparameter flow cytometry and CD45/SSC gating were used to analyze the surface and cytoplasmic antigen expressions in 115 cases of AML. The results were compared with the French-American-British (FAB) Cooperative Group classification to help define the best use and role of multiparameter flow cytometry in the diagnosis and proper classification of AML. The results showed that CD38, CD38 and CD13 were the most commonly expressed antigen (94.8%, 91.3% and 89.6%, respectively). CD7 was the most commonly expressed lymphoid antigen (20.2%), followed by CD19 (16.5%) and CD2 (15%). Some immunophenotypes correlated with FAB type, including increased frequency of CD2 in M(3); lack of HLA-DR, CD34 and CD56 expression in M(3); increased frequency of CD19 in M(2), CD14 and CD56 in M(5) and lack of MPO in M(0). In conclusion, multiparameter flow cytometry is a reliable technique in the diagnosis of AML, and some immunophenotypes correlate with FAB type.
Immunophenotyping
Cytometry
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Objective Investigate and analyse the antigen expression and clinical significance in adult acute leukemia.Methods 45 patients with leukemia were determined with flow cytometry to obtain valid diagnosis.Results In 19 patients with acute lymphoblastic leukemia (ALL),there were 16.3% patients expressed myeloid antigen,which CD_(13) were 14.6% and CD_(33) 8.4%.The percent of CR(35.2%)in the ALL with positive myeloid antigen expression was significantly lower that (69.7%) with negative myeloid antigen expression.In 26 patients with acute myeloid leukemia (AML)there were 23.8% patients expressed lymphoblastic antigen,which CD_7 were 12.4% and CD_(19) 9.7%.The percent of CR(41.6%)in the AML with positive lymphoblastic antigen expression was lower that (62.1%) with negative lymphoblastic antigen expression.Conclusion The immunophenotyping could provide more reliable,sensitive and specific evidence in the clinical diagnosis and prognostic significance in acute leukemia.
Immunophenotyping
Clinical Significance
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To evaluate the immunophenotype of acute leukemia patients, the surface and cyto-plasmic antigen expression in 162 cases of acute leukemia were analyzed by multiparameter flow cytometry and CD45/SSC gating. The results showed that CD117 (94. 9%),CD13 (88. 5%) and CD33(70. 5%) were mainly expressed in ANLL patients; cCD79a(100%) ,CD19(92. 1%) were chiefly expressed in B-ALL patients, and in T-ALL patients, cCD3(100%) and CD2(83. 3%) were expressed; For the expression of lymphoid differentiation antigen Ly+ANLL, CD7 (56. 2%) and CD19(31. 2%) were chiefly found, and for myeloid antigen My+ALL, CD13(88. 9%) and CD33(27. 8%) were detected. In conclusion, multiparameter flow cytometry and three-color direct immunofluorescence staining methods may be of important clinical significance in diagnosis, therapy and prognosis of acute leukemia.
Immunophenotyping
CD117
Cytometry
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Objective To explore the expression of cell surface antigen CD117 in different types of acute leukemia (AL) and its significance. Methods The expression of CD117 in bone marrow or peripheral blood of 139 AL patients were detected by flow cytometry, then analyzed and compared with each other. Results There was no significant difference in expression level of CD+117 cell between acute lymphoid leukemia (ALL) patients and normal control(P 0.05). The CD+117 patient rate and the expression level of CD+117 cell in acute myeloid leukemia (AML) patients were significantly higher than those in ALL patients and normal control(P 0.05). In AML subtypes, the expression of CD117 was the highest in M0/M1 and the lowest in M5b. Among 5 membrane antigen markers, the sensitivity of CD117 in detecting AML was lower than that of CD13 and CD33, but the specificity of CD117 was higher than the latter. The expression of CD117 in AML was significantly correlated with the expression of CD34 and HLA-DR (P 0.01), but the expression of CD117 in M3 was obviously higher than the latter. Conclusion CD117 can be used as the important immunophenotype indicator in MIC diagnosis and typing for AL patients, it has an important significance for diagnosing AML and distinguishing AML from ALL.
CD117
Immunophenotyping
Clinical Significance
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To study the immunophenotyping of adult patients with acute leukemia and its association with the prognosis.Immunophenotyping was performed in 106 adult patients with acute leukemia by three-color flow cytometry analysis using CD34/SSC gating.The antigens expressed in 71 patients with acute myeloid leukemia (AML) were mainly CD13, CD33, HLA-DR, CD34 and CD117. Lymphoid antigen expression was identified in 23.9% adult AML patients and CD56 antigen expression in 15.5% of the AML patients. In 29 patients with acute lymphoblastic leukemia (ALL), the expressed antigens were mainly HLA-DR, CD10, CD19, CD34 and CD7, and 34.5% of these patients were found to be positive for myeloid antigen expression. Complete remission (CR) rate in AML patients with lymphoid antigen expression after chemotherapy was lower than that in AML patients without lymphoid antigen expression (52.9% vs 77.8%, P<0.05), and no significant impact was noted of myeloid antigen expression in ALL on the CR of the patients (70.0% vs 94.7%, P>0.05). The CR rate in AML with CD56 antigen expression was lower than that in AML without CD56 expression (36.4% vs 78.3%, P<0.025), and the CR rate in CD34+ AML was lower than that in CD34- AML (56.0% vs 80.4%, P<0.05).Gating of CD45/SSC can eliminate the interference of normal cells to render more reliable immunophenotyping results. The expressions of CD56+, CD34+, lymphoid antigen in adult AML patients, who have lower CR rate, often signify poor prognosis.
Immunophenotyping
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Objective To explore the significance of immunophenotype by flow cytometry in the diagnosis and treatment of acute leukemia(AL).Methods Flow cytometry and CD45/side scatter(SSC) gating were used to analyze antigenic expressions in 93 patients with AL.Results The antigens expressed in 69 patients with acute myeloid leukemia(AML) were mainly CD13,CD33,HLA-DR and CD117;lymphoid antigens expression was identified in 20.2% of AML patient,there was a significant difference between CD7+ and CD7-in the percent CR(P0.05).In 13 patients with acute B-lymphoblastic leukemia,the expressed antigens were mainly CD19,CD34,HLA-DR,CD20,CD22,and 31% of these patients were found to be positive for myeloid antigen expression.There was no significant difference between CD13+ and CD13-in the percent CR(P0.05).The antigens expressed in 7 patients with acute T-lymphoblastic leukemia were mainly CD5,CD3,CD7,and CD34.Conclusion Immunological classification by flow cytometry has important value in therapy and prognosis of AL.
Immunophenotyping
CD5
CD117
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CD45 or LCA (leukocyte common antigen) is transmembrane glycoprotein active as tyrosine kinase. It is expressed solely on precursor and mature hematopoietic cells. Its significance is derived in the possibility of differentiate between hematopoietic (CD45+) and nonhematopoietic (CD45-) tissue, especially in the context of the differential diagnosis between extranodal lymphomas and nodal and bone nonhematologic malignancies and their metastases. In the immunophenotyping of acute leukemia, CD45 is a biomarker used to distinguish the population of blasts, characterized by the expression of low CD45 antigen and moderate to high side scrater (SSC) parameter. The CD45 antigen immunophenotyping is currently defined and confirmed by numerous data in the detection of minimal residual disease (MRD) in acute lymphoblastic leukemia (ALL), especially in CD45 negative types. Also, its usage contributes to higher sensitivity of multiparametric flow cytometry in detecting MRD of acute myeloid leukemia (AML). Due to the variable expression of CD45RO and CD45RA isoforms during normal hematopoiesis, the expression of these isoforms on the blasts varies based on the type of AML and between B-cell and T-cell ALL. The subgroup of CD45 negative leukemias is defined by the expression of this antigen 75%) of CD45 and moderate (>20%) intensity of CD20 antigen are independent prognostic factors in ALL. Recent studies in childhood AML have described that the high percentage of CD34+/CD38-/CD45- blasts is associated with worse prognosis. Meanwhile, the research on CD45 antigen significance in adult forms of leukemia remains scarce. Only few papers have shown the possible positive prognostic features of CD45- negative ALL, while we have not been able to find the data on CD45 expression in AML. In our study, 128 patients suffering from acute leukemia were included. Ninety-seven (75.8%) patients were diagnosed with AML, 26 patients (20.3%) with ALL, four cases of biphenotypic leukemias (AML/ALL) and one case of undifferentiated leukemia were recorded. Only one cases in ALL subgroups was CD45 negative, while additional four cases had moderate CD45 expression intensity ( 75%), with only six subjects having the CD45 intensity <90%. Based on our data, it seems that the prevalence of CD45 negative leukemia subgroup in adulthood is lower than in those in children. Due to the small number in this sample, we were not able to analyse further its prognostic significance.
Immunophenotyping
Minimal Residual Disease
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