title: A simple one-tube assay for immunophenotypical quantification of leukemic stem cells in acute myeloid leukemia.
abstract: Relapses after initial successful treatment in acute myeloid leukemia are thought to originate from the outgrowth of leukemic stem cells. Their flow cytometrically assessed frequency is of importance for relapse prediction and is therefore assumed to be implemented in future risk group profiling. Since current detection methods are complex, time- and bone marrow consuming (multiple-tubes approach), it would be advantageous to have a broadly applicable approach that enables to quantify leukemia stem cells both at diagnosis and follow-up. We compared 15 markers in 131 patients concerning their prevalence, usefulness and stability in CD34(+)CD38(-) leukemic stem cell detection in healthy controls, acute myeloid leukemia diagnosis and follow-up samples. Ultimately, we designed a single 8-color detection tube including common markers CD45, CD34 and CD38, and specific markers CD45RA, CD123, CD33, CD44 and a marker cocktail (CLL-1/TIM-3/CD7/CD11b/CD22/CD56) in one fluorescence channel. Validation analyses in 31 patients showed that the single tube approach was as good as the multiple-tube approach. Our approach requires the least possible amounts of bone marrow, and is suitable for multi-institutional studies. Moreover, it enables detection of leukemic stem cells both at time of diagnosis and follow-up, thereby including initially low-frequency populations emerging under therapy pressure.
authors.affiliation: Department of Hematology, VU University Medical Center, Cancer Center Amsterdam, Amsterdam, The Netherlands.
authors.forename: W
authors.initials: W
authors.lastname: Zeijlemaker
authors.affiliation: Department of Hematology, VU University Medical Center, Cancer Center Amsterdam, Amsterdam, The Netherlands.
authors.forename: A
authors.initials: A
authors.lastname: Kelder
authors.affiliation: Department of Hematology, VU University Medical Center, Cancer Center Amsterdam, Amsterdam, The Netherlands.
authors.forename: Y J M
authors.initials: YJ
authors.lastname: Oussoren-Brockhoff
authors.affiliation: Department of Hematology, VU University Medical Center, Cancer Center Amsterdam, Amsterdam, The Netherlands.
authors.forename: W J
authors.initials: WJ
authors.lastname: Scholten
authors.affiliation: Department of Hematology, VU University Medical Center, Cancer Center Amsterdam, Amsterdam, The Netherlands.
authors.forename: A N
authors.initials: AN
authors.lastname: Snel
authors.affiliation: Department of Hematology, VU University Medical Center, Cancer Center Amsterdam, Amsterdam, The Netherlands.
authors.forename: D
authors.initials: D
authors.lastname: Veldhuizen
authors.affiliation: Department of Hematology, VU University Medical Center, Cancer Center Amsterdam, Amsterdam, The Netherlands.|Department of Pediatric Oncology/Hematology, VU University Medical Center, Amsterdam, The Netherlands.
authors.forename: J
authors.initials: J
authors.lastname: Cloos
authors.affiliation: Department of Hematology, VU University Medical Center, Cancer Center Amsterdam, Amsterdam, The Netherlands.
authors.forename: G J
authors.initials: GJ
authors.lastname: Ossenkoppele
authors.affiliation: Department of Hematology, VU University Medical Center, Cancer Center Amsterdam, Amsterdam, The Netherlands.
authors.forename: G J
authors.initials: GJ
authors.lastname: Schuurhuis
mesh_terms: D051997:ADP-ribosyl Cyclase 1 / Q000032:analysis; D018952:Antigens, CD34 / Q000032:analysis; D006801:Humans; D016130:Immunophenotyping; D053650:Interleukin-3 Receptor alpha Subunit / Q000032:analysis; D015470:Leukemia, Myeloid, Acute / Q000473:pathology*; D008562:Membrane Glycoproteins / Q000032:analysis; D014411:Neoplastic Stem Cells / Q000276:immunology*; D063268:Sialic Acid Binding Ig-like Lectin 3 / Q000032:analysis
chemical_list: D018952:Antigens, CD34; C571163:CD33 protein, human; C115022:IL3RA protein, human; D053650:Interleukin-3 Receptor alpha Subunit; D008562:Membrane Glycoproteins; D063268:Sialic Acid Binding Ig-like Lectin 3; C498154:CD38 protein, human; D051997:ADP-ribosyl Cyclase 1