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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.
AuthorsAndreas Kremser, Stefanie Kufner, Elke Konhaeuser, Tanja Kroell, Andreas Hausmann, Johanna Tischer, Hans-Jochem Kolb, Horst Zitzelsberger, Helga Schmetzer
JournalLeukemia & lymphoma (Leuk Lymphoma) Vol. 54 Issue 6 Pg. 1297-308 (Jun 2013) ISSN: 1029-2403 [Electronic] United States
PMID23163701 (Publication Type: Journal Article)
Topics
  • Cell Differentiation (immunology)
  • Cells, Cultured
  • Dendritic Cells (immunology, metabolism)
  • Humans
  • Immunophenotyping
  • Immunotherapy
  • In Situ Hybridization, Fluorescence
  • Leukemia, Myeloid, Acute (genetics, immunology, metabolism, therapy)
  • Lymphocyte Activation
  • T-Lymphocyte Subsets (immunology, metabolism)

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