A. Symeonidis
Professor of Internal Medicine and Hematology, Head, Hematology Division, Dept of Internal Medicine at the University of Patras, President of the Hellenic (Greek) Myelodysplastic (MDS) and Bone Marrow Failure Syndromes' Study Group, Founder and Director of the Hellenic National MDS Registry, member of the Steering Committee of the European MDS Registry, Vice President of the Gaucher Disease Task Force of the European Hematology Association (EHA), member of 8 Medical Societies, including the Hellenic (Greek) Society of Hematology, ASH, EHA, EBMT, MDS Foundation and EWGGD and reviewer in 35 Scientific Journals. Expert in the field of bone marrow failure syndromes, B-cell malignancies, anemia
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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.
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.