A Novel Method for the Evaluation of Bone Marrow Samples from Patients with Pediatric B-Cell Acute Lymphoblastic Leukemia—Multidimensional Flow Cytometry

SIMPLE SUMMARY: By supporting the selection of the most suitable treatment protocol, the advancement of diagnostic methods contributes to achieving the best possible outcome for pediatric cases of acute lymphoblastic leukemia (ALL). In this study, we focused on a novel possibility in the flow cytometric (FC) analysis, as this method is the initial, crucial step in the diagnostic algorithm of ALL and can determine further diagnostic and therapeutic strategies. After the retrospective, multidimensional dot-plot-based FC analysis of 72 bone marrow samples of children with ALL, we found that the integrated appearance of immunophenotype resulted in a simple, quick, and accurate method. Furthermore, associations between immunophenotype and cytogenetic alterations were detected, which enabled the identification of cases with potential adverse outcome by completing the conventional FC analysis with multidimensional dot-plots. Standardized multi-center studies would be required to validate our results. ABSTRACT: Multicolor flow cytometry (FC) evaluation has a key role in the diagnosis and prognostic stratification of ALL. Our aim was to create new analyzing protocols using multidimensional dot-plots. Seventy-two pediatric patients with ALL were included in this single-center study. Data of a normal BM sample and three BM samples of patients with BCP-ALL were merged, then all B cell populations of the four samples were presented in a single radar dot-plot, and those parameters and locations were selected in which the normal and pathological cell populations differed from each other the most. The integrated profile of immunophenotype resulted in a simple, rapid, and accurate method. There were no significant differences between the percentages of lymphoblasts in the detection of minimal residual disease (MRD) by multidimensional or conventional FC method (p = 0.903 at Day 15 and p = 0.155 at Day 33). Furthermore, we found associations between the position and the number of clusters of blast cells in the radar plots and cytogenetic properties (p = 0.002 and p < 0.0001 by the position and p = 0.02 by the number of subclones). FC analysis based on multidimensional dot-plots is not only a rapid, easy-to-use method, but can also provide additional information to screen cases which require detailed genetic examination.