Immunotoxin BL22 induces apoptosis in mantle cell lymphoma (MCL) cells dependent on Bcl-2 expression
Christian Bogner
IIIrd Department of Medicine, Technical University of Munich, Munich
Search for more papers by this authorTobias Dechow
IIIrd Department of Medicine, Technical University of Munich, Munich
Search for more papers by this authorIngo Ringshausen
IIIrd Department of Medicine, Technical University of Munich, Munich
Search for more papers by this authorMichaela Wagner
IIIrd Department of Medicine, Technical University of Munich, Munich
Search for more papers by this authorMadlen Oelsner
IIIrd Department of Medicine, Technical University of Munich, Munich
Search for more papers by this authorGloria Lutzny
IIIrd Department of Medicine, Technical University of Munich, Munich
Search for more papers by this authorChristian Peschel
IIIrd Department of Medicine, Technical University of Munich, Munich
Search for more papers by this authorIra Pastan
Laboratory of Molecular Biology, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA
Search for more papers by this authorRobert J Kreitman
Laboratory of Molecular Biology, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA
Search for more papers by this authorThomas Decker
IIIrd Department of Medicine, Technical University of Munich, Munich
Onkologie Ravensburg, Ravensburg, Germany
Search for more papers by this authorChristian Bogner
IIIrd Department of Medicine, Technical University of Munich, Munich
Search for more papers by this authorTobias Dechow
IIIrd Department of Medicine, Technical University of Munich, Munich
Search for more papers by this authorIngo Ringshausen
IIIrd Department of Medicine, Technical University of Munich, Munich
Search for more papers by this authorMichaela Wagner
IIIrd Department of Medicine, Technical University of Munich, Munich
Search for more papers by this authorMadlen Oelsner
IIIrd Department of Medicine, Technical University of Munich, Munich
Search for more papers by this authorGloria Lutzny
IIIrd Department of Medicine, Technical University of Munich, Munich
Search for more papers by this authorChristian Peschel
IIIrd Department of Medicine, Technical University of Munich, Munich
Search for more papers by this authorIra Pastan
Laboratory of Molecular Biology, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA
Search for more papers by this authorRobert J Kreitman
Laboratory of Molecular Biology, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA
Search for more papers by this authorThomas Decker
IIIrd Department of Medicine, Technical University of Munich, Munich
Onkologie Ravensburg, Ravensburg, Germany
Search for more papers by this authorSummary
Mantle cell lymphoma (MCL) is an incurable mature B cell proliferation, combining the unfavourable clinical features of aggressive and indolent lymphomas. The blastic variant of MCL has an even worse prognosis and new treatment options are clearly needed. We analysed the effects of BL22, an immunotoxin composed of the Fv portion of an anti- CD22 antibody fused to a 38-kDa Pseudomonas exotoxin-A fragment on four MCL cell lines as well as on primary cells of four MCL patients. Apoptosis induction by BL22 was much more pronounced in MCL cell lines with low Bcl-2 expression (NCEB-1, JeKo-1 and JVM-2) compared to Granta-519 cells with high Bcl-2 expression. While the expression of the antiapoptotic protein Mcl-1 declined (NCEB-1, Granta-519), Bcl-2 levels remained unchanged in Granta-519 cells. However transfection of BCL2 cDNA into NCEB-1, JeKo-1 and JVM-2 cells significantly reduced BL22-mediated toxicity. Accordingly we examined the effects of Bcl-2 inactivation in Granta-519 cells using siRNA. Indeed, apoptosis induction was strongly enhanced in Granta-519 cells with silenced Bcl-2. Our results were confirmed in freshly isolated MCL-cells from patients with leukaemic MCL. We conclude that Bcl-2 expression is important for mediating resistance against the immunotoxin BL22 in MCL cells.
Supporting Information
Fig S1. (A) NCEB-1, NCEB-1/t BCL-2 cells, JeKo-1, JeKo-1/t-BCL2 cells as well as JVM-2 and JVM-2/tBCL2 cells were cultured in the presence of BL22 at the indicated concentration for 48 h. Phosphatidylserine exposure was measured using the annexin V assay. The mean percentage of viable cells (annexin – and PI- negative) ± SEM of three different experiments is shown. Paired t-tests were performed to test for differences between NCEB-1 and NCEB-1/tBCL2 cells (*), JeKo-1 and JeKo-1/tBCL2 cells (#), as well as JVM-2 and JVM-2/tBCL2 cells (+). */#/+ indicates when the difference between the cells was statistically significant (P < 0.05) (*: P 0.0217), (#: P 0.0175), (+: P 0.0408). (B) Bcl-2 expression in JVM-2 cells measured via FACS analysis. (C) Representative data of a CD22 expression analysis for JeKo-1, NCEB-1 and JVM-2 cells. Also shown are the isotype controls.
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