Abstract
Immunophenotypic analysis using multiparameter flow cytometry is an indispensable tool for diagnosis and management of acute leukemia. Mouse models have been widely used for medical research for more than 100 years and are indispensable for leukemia research. However, immunophenotypic analysis of murine leukemia was not always performed in published studies, and blast gating for isolation of blasts was shown only in very few studies. No systemic characterization of all types of murine acute leukemia in large cohorts by flow cytometry has been reported. In this study, we used flow cytometry to comprehensively characterize murine acute leukemia in a large cohort of mice. We found that murine T-lymphoblastic leukemia/lymphoma (T-ALL) exhibits a distinctive “blast gate” (CD45bright) with CD45/side scatter gating that differs from the “blast gate” (CD45dim) of human T-ALL. By contrast, murine B-lymphoblastic leukemia and acute myeloid leukemia show the same blast region (CD45dim) as human leukemia. Using blast cell gating, we for first time detected T-ALL development in FLT3-ITD knock-in mice (incidence: 23%). These leukemic cells were selectively killed by the FLT3 inhibitors crenolanib and midostaurin in vitro. These data suggest that FLT3-ITD plays a potential role in the pathogenesis of T-ALL and that FLT3-ITD inhibition is a therapeutic option in the management of patients with T-ALL. Our gating strategy for immunophenotypic analysis can be used for leukemogenesis and preclinical gene therapy studies in mice and may improve the quality of such analyses.
Highlights
The immunophenotypic analysis of acute leukemia by multiparameter flow cytometry is a powerful tool for proper identification of lymphoid or myeloid lineage and is indispensable in modern diagnosis and management of acute leukemia [1]
We found that murine T-lymphoblastic leukemia/ lymphoma (T-ALL) exhibits a distinctive “blast gate” (CD45bright) with CD45/side scatter gating that differs from the “blast gate” (CD45dim) of human T-ALL
Blasts from murine T-ALL were located in a gate with the highest CD45 fluorescence intensity (CD45bright) (Figures 2A-2P), whereas blasts from patients with T-ALL were exclusively present in the classic “blast gate” (cBG) (Figures 2C and 2D)
Summary
The immunophenotypic analysis of acute leukemia by multiparameter flow cytometry is a powerful tool for proper identification of lymphoid or myeloid lineage and is indispensable in modern diagnosis and management of acute leukemia [1]. CD45/side scatter (SS) gating for isolating blasts by flow cytometry was first proposed by Borowitz and Stelzer [2, 3] and is widely used. In mouse models expressing transgene and/or marker genes such as enhanced green fluorescent protein (EGFP) in hematopoietic cells, leukemic cells may be isolated by flow cytometry [5, 10,11,12,13]. No systemic characterization of all types of murine acute leukemia in large cohorts by flow cytometry has been reported. We analyze murine acute leukemia by flow cytometry in a large cohort of mice. Our gating strategy for immunophenotypic analysis can be used for leukemogenesis and preclinical gene therapy studies in mouse models and may improve the quality of such analyses
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