title: Combined immunophenotyping and fluorescence in situ hybridization with chromosome-specific DNA probes allows quantification and differentiation of ex vivo generated dendritic cells, leukemia-derived dendritic cells and clonal leukemic cells in patients with acute myeloid leukemia.
abstract: Antileukemic T-cell responses induced by leukemia-derived dendritic cells (DC(leu)) are variable, due to varying DC/DC(leu) composition/quality. We studied DC/DC(leu) composition/quality after blast culture in four DC media by flow cytometry (FC) and combined fluorescence in situ hybridization/immunophenotyping analysis (FISH-IPA). Both methods showed that DC methods produce variable proportions of DC subtypes. FISH-IPA is an elaborate method to study clonal aberrations in blast/DC cells on slides, however without preselection of distinct cell populations for FISH analysis. FISH-IPA data proved previous FC data: not every clonal/blast cell is converted to DC(leu) (resulting in various proportions of DC(leu)) and not every detectable DC is of clonal/leukemic origin. Preselection of the best of four DC methods for "best" DC/DC(leu) generation is necessary. DC(leu) proportions correlate with the antileukemic functionality of DC/DC(leu)-stimulated T-cells, thereby proving the necessity of studying the quality of DC/DC(leu) after culture. FC is the superior method to quantify DC/DC(leu), since a blast phenotype is available in every given patient, even with low/no proportions of clonal aberrations, and can easily be used to study cellular compositions after DC culture.
authors.affiliation: Department of Hematopoietic Cell Transplantations, University Hospital of Munich, Munich, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Kremser
authors.forename: Stefanie
authors.initials: S
authors.lastname: Kufner
authors.forename: Elke
authors.initials: E
authors.lastname: Konhaeuser
authors.forename: Tanja
authors.initials: T
authors.lastname: Kroell
authors.forename: Andreas
authors.initials: A
authors.lastname: Hausmann
authors.forename: Johanna
authors.initials: J
authors.lastname: Tischer
authors.forename: Hans-Jochem
authors.initials: HJ
authors.lastname: Kolb
authors.forename: Horst
authors.initials: H
authors.lastname: Zitzelsberger
authors.forename: Helga
authors.initials: H
authors.lastname: Schmetzer
mesh_terms: D002454:Cell Differentiation / Q000276:immunology*; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000276:immunology* / Q000378:metabolism*; D006801:Humans; D016130:Immunophenotyping; D007167:Immunotherapy; D017404:In Situ Hybridization, Fluorescence; D015470:Leukemia, Myeloid, Acute / Q000235:genetics / Q000276:immunology* / Q000378:metabolism* / Q000628:therapy; D008213:Lymphocyte Activation; D016176:T-Lymphocyte Subsets / Q000276:immunology / Q000378:metabolism