title: An in vitro model for cytogenetic conversion in CML. Interferon-alpha preferentially inhibits the outgrowth of malignant stem cells preserved in long-term culture.
abstract: IFN-alpha has been shown to prolong survival in chronic myeloid leukemia patients, but its mechanism of action is still not understood. The human cobblestone area-forming cell (CAFC) assay allows for the measurement of the concentration of normal as well as malignant stem cells, while their progeny can be measured in parallel long-term culture (LTC) in flasks. Using CAFC and LTC assays, we have examined direct effects of IFN-alpha (500; 5,000 IU/ml) on the maintenance and outgrowth of CD34-enriched normal and malignant stem cells, obtained from six patients with an established major cytogenetic response to IFN-alpha and from four nonresponding patients. CAFC concentrations were not affected by IFN-alpha. In contrast, IFN-alpha strongly inhibited the clonogenic output in flask LTC. Nucleated cells (NC) produced in LTC were evaluated by fluorescent in situ hybridization (FISH) for the presence of the Philadelphia (Ph) translocation. After 8 wk of LTC, the percentage of Ph+ NCs produced was significantly more inhibited by IFN-alpha in responding patients than in nonresponders. Control LTC without IFN-alpha showed no significant differences of Ph+ NC production between responders and nonresponders. These findings provide the first in vitro model for cytogenetic conversion and suggest that direct antiproliferative effects of IFN-alpha account for the cytogenetic response observed clinically.
authors.affiliation: Department of Hematology, Daniel den Hoed Cancer Center, Rotterdam, 3075 EA, The Netherlands. Cornelissen@hemh.azr.nl
authors.forename: J J
authors.initials: JJ
authors.lastname: Cornelissen
authors.forename: R E
authors.initials: RE
authors.lastname: Ploemacher
authors.forename: B W
authors.initials: BW
authors.lastname: Wognum
authors.forename: A
authors.initials: A
authors.lastname: Borsboom
authors.forename: H C
authors.initials: HC
authors.lastname: Kluin-Nelemans
authors.forename: A
authors.initials: A
authors.lastname: Hagemeijer
authors.forename: B
authors.initials: B
authors.lastname: Löwenberg
mesh_terms: D000328:Adult; D018952:Antigens, CD34 / Q000276:immunology; D002455:Cell Division / Q000187:drug effects*; D002467:Cell Nucleus / Q000473:pathology; D002478:Cells, Cultured; D002999:Clone Cells / Q000378:metabolism; D005260:Female; D005455:Fluorescent Antibody Technique; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D016898:Interferon-alpha / Q000494:pharmacology* / Q000627:therapeutic use; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000628:therapy*; D008297:Male; D008875:Middle Aged; D010677:Philadelphia Chromosome; D013234:Stem Cells / Q000187:drug effects*; D014178:Translocation, Genetic / Q000235:genetics
chemical_list: D018952:Antigens, CD34; D016898:Interferon-alpha