TY - JOUR

T1 - The granzyme B inhibitor PI-9 is differentially expressed in all main subtypes of pediatric acute lymphoblastic leukemias

AU - Classen, Carl Friedrich

AU - Ushmorov, Alexey

AU - Bird, Phillip

AU - Debatin, Klaus Michael

PY - 2004/11/1

Y1 - 2004/11/1

N2 - 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.

AB - 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.

KW - Apoptosis

KW - Cytotoxicity

KW - Granzyme B

KW - Leukemia

KW - PI-9

UR - http://www.scopus.com/inward/record.url?scp=9144247731&partnerID=8YFLogxK

M3 - Article

C2 - 15531453

AN - SCOPUS:9144247731

VL - 89

SP - 1314

EP - 1321

JO - Haematologica

JF - Haematologica

SN - 0390-6078

IS - 11

ER -