Expression profile of orphan receptor tyrosine kinase (ROR1) and Wilms' tumor gene 1 (WT1) in different subsets of B-cell acute lymphoblastic leukemia
Mahdi ShabaniHossein Asgarian‐OmranParvaneh VossoughRamazan Ali SharifianMohammad FaranoushSoheila GhragozlouJalal KhoshnoodiAzam RoohiMahmood Jeddi‐TehraniHåkan MellstedtHodjatallah RabbaniFazel Shokri
46
Citation
55
Reference
10
Related Paper
Citation Trend
Abstract:
Recent molecular investigations have demonstrated over-expression of a large number of tumor associated antigens (TAAs) in a variety of malignancies. Over-expression of ROR1 gene, a member of the receptor tyrosine kinase family, has recently been reported in B-cell chronic lymphocytic leukemia. Wilms' tumor gene 1 (WT1) has long been known as a universal TAA expressed in a variety of solid and hematopoietic malignancies. In the present study, the expression profile of ROR1 and WT1 was investigated in different immunophenotypic subsets of B-cell acute lymphoblastic leukemia (B-ALL) patients. RT-PCR method was used to determine the ROR1 and WT1 genes expression in bone marrow (BM) and peripheral blood (PB) samples from 51 newly diagnosed Iranian B-ALL patients. Isolated tumor cells from all patients were immunophenotyped by flow cytometry. Based on immunophenotypic results, our B-ALL patients were classified in four differentiation subsets; Pro-B (n = 7), Pre-B I (n = 29), Pre-B II (n = 13) and Immature/mature B-ALL (n = 2). Although ROR1 was over-expressed in more mature subsets (16.7%, 42.9%, 45.5% and 100%, respectively), WT1 was more represented in immature subsets of B-ALL patients (57.1%, 64.3%, 38.5% and 0%, respectively). Comparison of the frequency of ROR1 and WT1 positive samples at each immunophenotypic subtype revealed statistically significant difference only in Pre B I subtype (p = 0.02). Our results suggest that expression of ROR1 and WT1 in B-ALL is associated with the differentiation stage of the leukemic cells.Keywords:
ROR1
Immunophenotyping
Objective To explore immunophenotyping specificities in acute leucocythemia via flow cytometry.Methods Flow cytometry was used to have immunophenotyping of acute leucocythemia tested and assist in making the diagnosis leukemia detection.Results(1)Acute lymphocytic leukemia was divided into T cells in series of acute leukemia(T-ALL) and B cell lines acute leukemia(B-ALL) by using specific antibodies.(2)with specific antibodies,acute myelogenous leukemia(AML) can be diagnosed more clear.(3)Acute lymphocytic leukemia(ALL) with myeloid antigen expression(My-ALL),AML antigen expression from Ly(AML) and mixed leukemia were found.Conclusion Immunophenotype can reflect the heterogeneity of leukemia cells more distinctly and consequently,enhance the exactness of diagnosis.
Immunophenotyping
Acute lymphocytic leukemia
Cite
Citations (0)
In order to identify neuroblastoma cells infiltrating the bone marrow, a triple-color flow-cytometric assay was developed combining CD56 and CD45 with the intracellular anti-NB84 specific antibody.The bilateral aspirates obtained from 27 consecutive children over the age of one year with stage 4 neuroblastoma were evaluated.Neuroblastoma cells were detected in the bone marrow of 17/27 (63%) and 19/27 (70%) cases using cytomorphology and triple-color flow-cytometry, respectively. Using cytometry, the percentage of CD56+/NB84+/CD45-cells infiltrating the bone marrow ranged from 0.02% to 65%. Five out of eight patients without bone marrow involvement according to cytometry are in continuous complete remission, while only 3 out of 19 patients whose bone marrow gave positive results are still alive.By combining CD45 and CD56 with the specific antibody, NB84, directed against neuroblastoma cells, we developed a rapid and reliable cytometric assay that can be associated with conventional cytomorphological bone marrow evaluation to detect infiltrating neuroblastoma cells, especially in cases of dubious positivity.
Immunophenotyping
Cytometry
Cite
Citations (19)
Objective It is to study the method and clinical significance of acute leukemia immunophenotyping by multi-parameter flow cytometry.Methods Immunophenotyping analysis was done by three color flow cytometry with the method of CD_(45)/SSC double parameter and scatter spot picture.Results In the 79 cases of leukemia,42 cases expressed medullary system antigen in which CD_(33) expression was the highest,12 cases(28%) with CD7 expression,2 cases(5%) with CD_2 expression,2 cases(5%) with CD_3 expression,2 cases(5%) with CD_(19) expression.39 cases expressed lymphocyte system antigen in which CD_(10) expression was the highest,6 cases(6%) with CD_(33) expression,4?cases(10%) with CD_(64) expression,2 cases was double expression leukemia.Conclusion Multi-parameter flow cytometry can accurately type acute leukemia,so it has an important value for the treatment choice and prognosis diagnosis of the patients with leukemia.
Immunophenotyping
Cytometry
Cite
Citations (0)
Objective To describe the procedure to perform three - color flow cytometric analysis on lysed whole bone marrow preparations for acute and chronic leukemia. Methods This approach was used to 115 patients with acute leukemia and 10 patients of chronic leukemia. Leukemia cells were analyzed by dual -parameter flow cytometry to distinguish normal lymphocytes, monocytes, granulocytes and nucleated red blood cells with combination of CD45 intensity and side scatter. Results The estimate of blasts by flow cytometric analysis was correlated highly with morphologic leukemia cell counts over a wide range. The pattern on the CD45/SS display varied in different FAB subtypes of leukemia. When CD45 - PCS was combined with FITC - and PE - conjugated antibodies, it was possible to set a gate on leukemia blasts and demonstate the dual -parameter analysis of the expressions of two additional markers in the leukemia population. Furthermore, this method also showed pattern display on the specimens of remission satisfactorily . Conclusion This gating strategy was superior to traditional FS versus SS display in immunophenotyping of leukemia.
Immunophenotyping
Cytometry
Cite
Citations (0)
The aim of this study was to evaluate the heterogeneity of immunophenotype features in acute leukemia patients and to detect the presence of leukemia-associated immunophenotypes. We prospectively investigated the phenotype of blast cells from 44 adult acute leukemia patients using a large panel of monoclonal antibodies by multiparametric flow cytometry. Thirty-three patients were classified as AML according to the FAB classification. Eleven patients were diagnosed as ALL (10 cases B-ALL, 1 case T-ALL) according to both FAB and immunnophenotyping. We found leukemia-associated phenotypes in 28 of 33 AML patients (84.8%) and in 8 of 11 ALL patients (72.7%). In 61.1% of patients more than one aberrant phenotype was observed. Linear infidelity was the most frequent aberrancy in both AML (64.3%) and ALL (37.5%) subgroups. The present study shows that MFC is a helpful method for sufficient identification of leukemic cells and for determination of blast cells immunophenotype heterogeneity. The double stain flow cytometry in our study revealed aberrant phenotypes in up to 81.8% patients.
Immunophenotyping
Stain
Cite
Citations (8)
In humans and in mouse models, precursor B-cell lymphoblastic leukemia (B-ALL)/lymphoblastic lymphoma (B-LBL) can be classified as either the pro-B or pre-B subtype. This is based on the expression of antigens associated with the pro-B and pre-B stages of B-cell development. Antigenic markers can be detected by flow cytometry or immunohistochemistry (IHC), but no comparison of results from these techniques has been reported for murine B-ALL/LBL. In our analysis of 30 cases induced by chemical or viral mutagenesis on a WT or Pax5 +/– background, 18 (60%) were diagnosed as pro-B by both flow cytometry and IHC. Discordant results were found for 12 (40%); 6 were designated pro-B by IHC and pre-B by flow cytometry and the reverse for the remaining 6 cases. Discordance occurred because different markers were used to define the pro-B–to–pre-B transition by IHC vs flow cytometry. IHC expression of cytoplasmic IgM (μIgM) defined the pre-B stage, whereas the common practice of using CD25 as a surrogate marker in flow cytometry was employed here. These results show that CD25 and μIgM are not always concurrently expressed in B-ALL/LBL, in contrast to normal B-cell development. Therefore, when subtyping B-ALL/LBL in mice, an IHC panel of B220, PAX5, TdT, c-Kit/CD117, CD43, IgM, and ΚLC should be considered. For flow cytometry, cytoplasmic IgM may be an appropriate marker in conjunction with the surface markers B220, CD19, CD43, c-Kit/CD117, BP-1, and CD25.
Immunophenotyping
CD43
CD117
CD5
B-cell lymphoma
Cite
Citations (3)
Objective:To discuss the feasibility and significance of the CD 45 gating in the immunophenotype of acute leukemia by flow cytometry.Methods:Using the CD 45 monoclonal antibody and flow cytometry,the cells from 32 peripheral bloods of healthy adults and 56 bone marrows from patient diagnosed as acute leukemia were examined and analysed.Results:With the healthy group versus acute leukemia group,the percent and the mean fluorescence intensity of CD 45 FITC positive cells in lymphocyte gate ( ±s ) were separately:(99.1±0.5)% vs (85.2±9.3)%( P 0.05) and (31.66±8.48) vs (2.46±1.07) ( P 0.001);CD 45 PE were (99.3±0.7)% vs (90.2±6.6)%( P 0.05) and 106.99±25.94 vs 13.86±9.51( P 0.001).After mixing the leukemic cells in white blood cells as 1∶1 000,CD 45 and side scatter easily gate on blast cells.Conclusion:CD 45 gating can simply and clearly distinguish a few blast and immature leukemic cells from the peripheral blood.The sensitivity was 10 -3 .
Immunophenotyping
Cytometry
Cite
Citations (0)
Summary Flow cytometric identification of small numbers of precursor B‐cell acute lymphoblastic leukaemia (B‐ALL) cells in post‐treatment marrow specimens could benefit from the identification of additional, easily detectable markers that could be used in most cases. In this study, we evaluate whether bcl‐2 expression quantified by four‐colour flow cytometry can be effectively used to discriminate precursor B‐ALL blasts from normal B‐cell precursors (haematogones) and function as a leukaemia‐specific marker. Levels of bcl‐2 in the 22 precursor B‐ALL cases studied were found to be significantly higher (over sixfold higher on average) than those present in haematogone populations from 22 control marrow specimens. Higher relative levels of bcl‐2 expression in the B‐ALL cases were maintained with respect to both immature CD34 + and more mature CD34 − haematogone subsets, and appeared stable. Dilutional studies indicated that multiparameter flow cytometry analysis using bcl‐2 could identify precursor B‐ALL blasts representing as few as 1% of CD19 + cells or 0·01% of total leucocytes in bone marrow specimens containing substantial numbers of haematogones. This study suggests that bcl‐2 may be an important marker for flow cytometric detection and quantification of small numbers of residual precursor B‐ALL cells in bone marrow specimens.
Acute lymphocytic leukemia
Cite
Citations (18)
Immunophenotyping
Hematology
Cite
Citations (2)