Immunophenotypic analysis of myelodysplastic syndromes (original) (raw)
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Cytometry Part B-clinical Cytometry, 2010
A heterogeneous spectrum of immunophenotypic abnormalities have been reported in myelodysplastic syndromes (MDS). However, most studies are restricted to the analysis of CD34+ cells and/or other major subsets of CD34− cells, frequently not exploring the diagnostic and prognostic impact of immunophenotyping.A heterogeneous spectrum of immunophenotypic abnormalities have been reported in myelodysplastic syndromes (MDS). However, most studies are restricted to the analysis of CD34+ cells and/or other major subsets of CD34− cells, frequently not exploring the diagnostic and prognostic impact of immunophenotyping.Methods:We propose for the first time an immunophenotypic score (IS) based on the altered distribution and immunophenotypic features of maturing/mature compartments of bone marrow (BM) hematopoietic cells in 56 patients with MDS that could contribute to a refined diagnosis and prognostic evaluation of the disease.We propose for the first time an immunophenotypic score (IS) based on the altered distribution and immunophenotypic features of maturing/mature compartments of bone marrow (BM) hematopoietic cells in 56 patients with MDS that could contribute to a refined diagnosis and prognostic evaluation of the disease.Results:Although MDS-associated phenotypes were detected in reactive BM, the overall immunophenotypic profile of BM cells allowed an efficient discrimination between MDS and both normal and reactive BM, once the number and degree of severity of the abnormalities detected per patient were simultaneously considered in the proposed IS. Interestingly, increasingly higher IS were found among patients with MDS showing adverse prognostic factors and in low- versus high-grade cases. The most informative prognostic factors included the number of CD34+ cells, presence of aberrant CD34−/CD117+ precursors, decreased mature neutrophils and CD34− erythroid precursors, and increased numbers of CD36−/lo erythroid precursors; in addition, the IS was an independent prognostic factor for overall survival.Although MDS-associated phenotypes were detected in reactive BM, the overall immunophenotypic profile of BM cells allowed an efficient discrimination between MDS and both normal and reactive BM, once the number and degree of severity of the abnormalities detected per patient were simultaneously considered in the proposed IS. Interestingly, increasingly higher IS were found among patients with MDS showing adverse prognostic factors and in low- versus high-grade cases. The most informative prognostic factors included the number of CD34+ cells, presence of aberrant CD34−/CD117+ precursors, decreased mature neutrophils and CD34− erythroid precursors, and increased numbers of CD36−/lo erythroid precursors; in addition, the IS was an independent prognostic factor for overall survival.Conclusions:Assessment of immunophenotypic abnormalities of maturing/mature BM cells allows an efficient discrimination between MDS and both normal and reactive BM, once the number and degree of severity of the abnormalities detected are simultaneously scored. Interestingly, progressively higher IS were found among patients with MDS with adverse prognostic features and shorter overall survival. © 2010 Clinical Cytometry SocietyAssessment of immunophenotypic abnormalities of maturing/mature BM cells allows an efficient discrimination between MDS and both normal and reactive BM, once the number and degree of severity of the abnormalities detected are simultaneously scored. Interestingly, progressively higher IS were found among patients with MDS with adverse prognostic features and shorter overall survival. © 2010 Clinical Cytometry Society
Issues in the Pathology of the Myelodysplastic Syndromes
Hematology/Oncology Clinics of North America, 1992
The article in this issue by Drs. Kouides and Bennett describes abnormalities of the hematopoietic cells in peripheral blood and bone marrow specimens of patients with myelodysplastic syndromes (MDS), and they review the criteria by which these features can be used for stratifying patients according to the French-American-British (FAB) classification scheme.6 Certainly, the application of these guidelines permits the identification of most patients with this bone marrow dyscrasia, and it allows them to be classified according to a system that has proved to have prognostic value. 30 • 108 However, a significant number of patients with MDS have blood and/or bone marrow findings that cause problems not only in classification, but even in allowing recognition that the patient has a myelodysplastic disorder."• 65 Problems in establishing the diagnosis of MDS may be due to a number of causes, Occasionally, these are related to failure to obtain adequate bone marrow aspirate smears for evaluation of cytology, either because of bone marrow hypocellularity or because of fibrosis of the marrow. In the first case, the unexpected finding of marrow hypocellularity and, thus, the lack of cells for detailed cytologic assessment may not permit an easy distinction between "hypocellular MDS" and aplastic anemia. In the second case, MDS with fibrosis of the marrow may not be readily separable from a variety of acute and chronic myeloid disorders that are also associated with marrow fibrosis and an inaspirable bone marrow. The diagnosis of additional cases may be difficult because they may exhibit unusual features that overlap with other neoplastic hematologic disorders. In particular, some chronic myeloproliferative disorders (CMPD) may mimic MDS because they not uncommonly demonstrate dyspoiesis as they transform to more accelerated stages. 45 • 68 • 71 • 85 Confusion between MDS
Leukemia Research, 2013
Developing a flow cytometric maturation/differentiation index of the bone marrow for the diagnosis of myelodysplastic syndromes (MDS) E. Verigou1, N. Smyrni1, G. Kolliopoulou1, E. Hala1, P. Lampropoulou1, G. Theodorou1, F. Kalogianni2, P. Zikos2, I. Starakis3, E. Solomou3, M. Karakantza1, A. Symeonidis1. 1Hematology Division Dept of Internal Medicine, Peripheral University Hospital of Patras, Patras, Greece; 2Hematology Division, St Andrew’s General Hospital of Patras, Patras, Greece; 3Internal Medicine, Peripheral University Hospital of Patras, Patras, Greece. Background: Establishing the diagnosis of MDS is a challenging task, due to disease heterogeneity. Morphology remains the gold standard, but dysplastic features can often be misleading and differential diagnosis of MDS and MDS-like cytopenias is intriguing. Bone marrow (BM) immunophenotype has not been incorporated in the diagnostic criteria of MDS, although it may be useful, especially in disputed cases of low-risk MDS. Introduction: We hypothesize that, BM myeloid cells exhibit immunophenotypic abnormalities, discriminating true from pseudoMDS, as well as maturation and differentiation blocks rather than marrow failure, and created a parametric index of BM normal granulopoietic capacity. Purpose: We aimed to establish a BM maturation/differentiation index, thus maximi-zing the diagnostic utility of Flow Cytometry data, and simplify their interpretation, by quantifying antigenic patterns, based on mathematical modeling rather than, on conventional sequential biparametric analysis. Materials and Methods: BM samples from 104 subjects were analysed for CD45PC7, CD11bPC5, CD16FITC and CD13PE expression (Beckman Coulter, FC500 flow cytometer). Sixty-eight patients had MDS (40 low risk, 28 high risk) and 26 patients had another diagnosis (ITP, chronic idiopathic neutropenia, systemic lupus erythema-tosus, LGL leukemia, age-related cytopenias, aplastic anemia, myelofibrosis etc). Moreover, 10 BM samples of patients with post-MDS acute myeloid leukemia (AML) were analyzed. Results: CD16 and CD11b antigen expression pattern was chosen for their reproduci-bility and biological significance. Combining the percentage ratio of the regions O (maturing and differentiated myeloid cells) and N (myeloid precursors/ immature cells, Fig. 1), with a 3-dimensional space representing the distribution of maturing cells in a 3D plot of fluorescence levels of CD16, CD11b and CD45, we resulted in HeSK* ratio as follows: x CD11b 0 y C D16 0 z C D45 neutro p0 10^6 pN where x is the median of CD11b in region O, y is the median of CD16 in region O, z is the median of CD45 in region neutro, pO is the percentage of region O in the total CD11b/CD16 diagram gated in neu Figure 2. HeSK/(%) blasts. tro, pN is the percentage of region N in the total CD11b/CD16 gated in neutro and 106 is an empirical parameter. The ratio could quantify the abnormal differentiation profile of maturing myeloid cells and distinguish MDS from non-MDS samples with high statistical significance (Kruskal-Wallis test, p<0.0001, Fig. 2). Descriptive statistics are shown in Table 1. Conclusions: HeSK correlated with morphological findings and highlights cases of clinical interest. Our future goal is to examine the behavior of antigen expression (as captured in HeSK) before and after therapy, estimate the prognostic value of the ratio and amplify its diagnostic utility. *HeSK: Authors’ name acronyms.
Leukemia Research, 2006
Recent studies concerning the pathophysiology of myelodysplastic syndromes (MDS) have shown evidences for the existence of complex interactions between hematopoietic stem cells and the bone marrow (BM) microenvironment. We analyzed the B-lymphocyte maturation in BM of patients with MDS. For this purpose, 41 newly-diagnosed patients were analyzed. Enumeration and characterization of CD34+ and CD34− B-cell precursors and mature B-lymphocytes was performed using multiparameter flow cytometry. BM from eight transplant donors and six orthopedic surgery patients were used as controls. CD34+/CD45 lo B-cells were found in 17/22 patients with RA/RARS and in 5/13 with RAEB. In patients with RAEB-t and CMML no CD34+ B-cell precursors could be detected. A positive correlation was found between CD34+ and CD34− B-cell precursors (r = 0.52). CD34+ B-cell precursors presented an inverse correlation with BM percentage of blasts and peripheral leukocytes and a positive one with hemoglobin. Asynchronous antigen expression (CD19+/CD79a− cells) was found in 7/11 cases of RA/RARS and 6/18 cases of RAEB in which this phenotype was examined. Abnormal patterns of expression for at least one antigen was found in 91% of RA/RARS cases and in 74% of RAEB. Underexpression of TdT and CD79a were the most frequent abnormalities. Our results present evidences of an abnormal B-cell maturation in MDS. This may be an evidence that B-lymphocytes are derived of the abnormal clone. But it may also be the consequence of influences of abnormalities of BM microenvironment leading to an impaired commitment and maturation of the B-cell line in MDS. Studies performed with purified well-characterized B-cells may further elucidate these abnormalities.
Leukemia Research, 2007
The diagnosis of myelodysplastic syndromes (MDS) is based on peripheral cytopenias, bone marrow (BM) morphology and karyotyping. This may be difficult in cases with few dysplastic elements in BM and a normal karyotype. We examined the utility of flow cytometric analysis for the differential diagnosis between MDS and non-clonal disorders (NCD) presenting peripheral cytopenias. Quantitative assessment of CD45, CD16, CD13, CD11b, CD10 and CD64 in granulocytes and monocytes, and CD71 and glycophorin A in erythroblasts besides CD34+ cell count was performed in BM of 31 consecutive newly diagnosed patients with MDS, 11 patients with NCD and 11 healthy controls (BM donors). In MDS, the median number of phenotypic abnormalities found was 3 (1-8). The WPSS score showed a correlation with the total number of changes per case (r = 0.48; p = 0.002). Decreased SSC in promyelocytes correlated with the peripheral neutrophil count (r = −0.46; p = 0.007). In NCD, the normal variation of antigen expression along granulocytic and erythroblast maturation was always maintained. In the discriminant analysis, SSC of CD34+ cells, together with that of promyelocytes and metamyelocytes were able to correctly classify 87% of the cases as clonal or non-clonal. Our quantitative approach permitted to detect at least one abnormality in antigen expression in every case of MDS. However, the most important parameters for differential diagnosis with NCD were the analysis of the granularity in immature cells, especially of the granulocytic series.
Bone marrow morphology and classification systems in myelodysplastic syndromes
Cancer Treatment Reviews, 2007
One of the hallmarks of myelodysplastic syndromes (MDS) is their morphological heterogeneity which complicates the diagnosis and classification of these bone marrow disorders. The French-American-British (FAB) classification system which is based on cytological findings and the blast cell percentage in the bone marrow (BM) and peripheral blood (PB) was the universally accepted reference standard for categorization of MDS samples from 1982 to 1999 when it was modified by an expert panel of the World Health Organization (WHO). The WHO classification system distinguishes eight subgroups of MDS and is based on three principles: distinction between unilineage and multilineage dysplasia of haematopoietic cells, definition of refined blast cell percentages in BM and PB for diagnosing patients with RAEB and AML, as well as incorporation of genetic data into the classification scheme. When compared to the FAB proposals, the WHO classification appears to define more homogeneous subgroups of patients and to provide greater prognostic power. However, prospective studies of large patient populations will be required to decide whether the WHO classification allows accurate risk assessment and facilitates clinical decision-making for individual MDS patients.
Cancer, 2009
BACKGROUND: Because of limited reproducibility of morphologic features, the morphological categorization of initial myelodysplastic syndromes (MDS) cases remains a major task in a diagnostic setting. METHODS: To further evaluate the role of additional diagnostic methods for suspected early MDS, the authors analyzed 1965 cases with unclear cytopenia where at least cytomorphology and immunophenotyping were performed in parallel, combined with cytogenetics and molecular genetics. RESULTS: In 353 patients, both methods diagnosed malignant/nonmalignant disease other than MDS, and 557 patients had MDS-refractory anemia with excess of blasts/chronic myelomonocytic leukemia. The remaining 1055 patients (53.7%), where early MDS/reactive cytopenia had to be assumed, were categorized into 6 groups depending on cytomorphology/immunophenotyping results for or against MDS. In 659 of 1055 cases (62.4%) with suspected initial MDS, cytomorphology and immunophenotyping were concordant in the categorization of MDS/non-MDS. Cytogenetics, available in 951 of 1055 patients, revealed the highest frequency of aberrant karyotypes when both cytomorphology and immunophenotyping proposed MDS (63 of 227; 27.8%).
Medical Oncology, 2006
It has been recognized that some patients with myelodysplastic syndromes (MDS) develop immunologic abnormalities, but little is known of its correlations to MDS-specific disease features. In a retrospective study of 284 MDS patients, we identified 32 patients (11.3%) with clinical or serologic immunological abnormalities (group A) and compared them to the remaining 252 cases (group B). Group A consisted of 20 patients with clinical signs of autoimmune disease and 12 asymptomatic patients with serologic immunological abnormalities only. Apart from significant female predominance in group A (M/F = 2.5 vs M/F = 0.7, p = 0.001), the other clinical and biological features such as median age, distribution of MDS subtypes, incidence of karyotyopic abnormalities, "abnormal" in vitro growth of GM-progenitors and survival times were similar in the two groups. Autoimmune manifestations partially responded to immunosuppressive therapy, with moderate improvement of peripheral cytopenia. In addition, CD3 + , CD4 + , CD8 + , CD19 + , and CD56 + cells were quantified in peripheral blood of 38 patients. Matched with similarly aged healthy control group, most MDS patients showed significant lymphocytopenia, mainly due to the reduction of T-helper series (in both absolute numbers and percentage). B-cells were reduced in absolute numbers, but their percentage still overlapped with the control. No major abnormalities of natural killer cells (CD56 + ) were seen. We conclude that autoimmune diseases and asymptomatic immunologic abnormalities are common in patients with MDS, but except for female predominance, no correlation between these abnormalities and MDS-specific disease features were found.
Standards and impact of hematopathology in myelodysplastic syndromes (MDS)
Oncotarget, 2010
The diagnosis, classification, and prognostication of patients with myelodysplastic syndromes (MDS) are usually based on clinical parameters, analysis of peripheral blood and bone marrow smears, and cytogenetic determinants. However, a thorough histologic and immunohistochemical examination of the bone marrow is often required for a final diagnosis and exact classification in these patients. Notably, histology and immunohistology may reveal dysplasia in megakaryocytes or other bone marrow lineages and/or the presence of clusters of CD34-positive precursor cells. In other cases, histology may reveal an unrelated or co-existing hematopoietic neoplasm, or may support the conclusion the patient is suffering from acute myeloid leukemia rather than MDS. Moreover, histologic investigations and immunohistology may reveal an increase in tryptase-positive cells, a coexisting systemic mastocytosis, or bone marrow fibrosis, which is of prognostic significance. To discuss diagnostic algorithms, ...