title: Interferon-alpha directly inhibits DNA polymerase activity in isolated chromatin nucleoprotein complexes: correlation with IFN-alpha treatment outcome in patients with chronic myelogenous leukemia.
abstract: We have developed an in vitro assay to assess and predict the potential efficacy of in vivo interferon-alpha (IFN-alpha) treatment (5 x 10(6) units/m2 per day) for patients with chronic myelogenous leukemia (CML). Although determining the numbers and affinities of IFN-alpha receptors on CML cells has been developed as a method for predicting treatment response to IFN-alpha, it fails to predict response in CML. Previously, we and others observed that mitogens, toxins and lectins that bind to cell-surface receptors are endocytosed, escaping endosomes in order to act directly on cellular targets. Therefore, we tested the ability of low concentrations of IFN-alpha (5-10 units) to act directly on DNA polymerase (Pol) in purified chromatin nucleoprotein complexes (NPC). NPC were prepared by a methodology that uses direct treatment of leukocyte nuclei with MspI to generate six NPC-containing fractions (S1, M1, S2, M2, 0.1K and R). We found three general categories of in vitro DNA synthesis response for the six different NPC fractions isolated from the white blood cells of patients with CML (n = 19) before their treatment with IFN-alpha. IFN-alpha induced either stimulation, inhibition or had no apparent effect on Pol activity in the six different NPC fractions in a blind assay. In most of the NPC fractions isolated from the leukocytes of patients with progressive CML and in those from CML patients who failed to show a clinical response to IFN-alpha, this cytokine stimulated or had no effect on Pol activity.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Department of Tumor Biology, University of Texas M.D. Anderson Cancer Center, Houston 77030, USA.
authors.forename: N L
authors.initials: NL
authors.lastname: Nicolson
authors.forename: M
authors.initials: M
authors.lastname: Talpaz
authors.forename: G L
authors.initials: GL
authors.lastname: Nicolson
mesh_terms: D002458:Cell Fractionation; D002467:Cell Nucleus; D002843:Chromatin / Q000187:drug effects / Q000201:enzymology*; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D019003:Deoxyribonuclease HpaII; D015252:Deoxyribonucleases, Type II Site-Specific; D006801:Humans; D016898:Interferon-alpha / Q000494:pharmacology* / Q000627:therapeutic use*; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000201:enzymology / Q000628:therapy*; D007962:Leukocytes; D019384:Nucleic Acid Synthesis Inhibitors; D011237:Predictive Value of Tests; D016896:Treatment Outcome
chemical_list: D002843:Chromatin; D016898:Interferon-alpha; D019384:Nucleic Acid Synthesis Inhibitors; D004259:DNA-Directed DNA Polymerase; D019003:Deoxyribonuclease HpaII; D015252:Deoxyribonucleases, Type II Site-Specific