The granzyme B inhibitor PI-9 is differentially expressed in all main subtypes of pediatric acute lymphoblastic leukemias

Carl Friedrich Classen, Alexey Ushmorov, Phillip Bird, Klaus Michael Debatin

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18 Citations (Scopus)

Abstract

Background and Objectives. The success of bone marrow transplantation in leukemia depends on graft-versus-leukemia (GvL) effects, mediated by cytotoxic T lymphocytes (CTL) and natural killer (NK) cells. These act by CD95L or granule cytotoxins, such as granzyme B (GrB) whose only known inhibitor is proteinase inhibitor 9 (PI-9). Since PI-9 protects cells from CTL, PI-9 may counteract GvL in leukemias. Our aim was to establish methods to analyze the expression and function of PI-9. Design and Methods. We screened the most common pediatric ALL subsets, i.e. pre-B, common, and cortical T-ALL, for PI-9 expression, using a reverse transcription polymerase chain reaction (RT-PCR) approach which was correlated to semiquantitative and functional methods established in cell lines and patient probes. Results. In vitro, PI-9 mediated resistance towards CTL and inhibited GrB-induced clevage of caspase 3. In patient-derived ALL cells, PI-9 high and PI-9 low specimens were studied by flow cytometry, RT-PCR, in vitro GrB treatment and NK assay, demonstrating concordant results and PI-9-dependent target cell protection. Analysis of PI-9 in probes from ALL patients showed differential expression, but no correlation with immunotype. Interpretation and Conclusions. Our data suggest that PI-9 in pediatric ALL is differentially expressed, without close correlation to subtype. Since PI-9 considerably alters GrzB and killer cell sensitivity, it may strongly influence the efficacy of GvL effects. The approaches applied here will allow evaluation of the expression and function of PI-9 in larger series of malignancies.

Original languageEnglish
Pages (from-to)1314-1321
Number of pages8
JournalHaematologica
Volume89
Issue number11
Publication statusPublished - 1 Nov 2004

Keywords

  • Apoptosis
  • Cytotoxicity
  • Granzyme B
  • Leukemia
  • PI-9

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