Document Detail


Apoptosis induced by immunotoxins used in the treatment of hematologic malignancies.
MedLine Citation:
PMID:  10861457     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The recombinant immunotoxins anti-Tac(Fv)-PE38 (LMB-2), targeting the interleukin-2 receptor alpha subunit (IL-2Ralpha, Tac or CD25), and RFB4(dsFv)-PE38 (BL22), targeting CD22, are being evaluated in clinical trials as treatment for hematologic malignancies. The toxin moiety Pseudomonas exotoxin A (PE) of these recombinant molecules leads to the arrest of protein synthesis due to inactivation of elongation factor 2. Here, we provide evidence that cell lines derived from patients with hematologic malignancies react to immunotoxins not only with inhibition of protein synthesis but also with characteristic hallmarks of apoptosis such as caspase activation, cleavage of the "death substrate poly(ADP)-ribose polymerase and DNA laddering. Anti-Tac(Fv)-PE38 leads to a 10-fold increase in the cleavage of the fluorescent substrate DEVD-AFC, suggesting that a caspase-3-like enzyme is involved. This was verified by cleavage of caspase-3 (CPP32). MT1 cells exhibited DNA laddering after treatment with immunotoxin, which was reversed by pre-treatment with the protease inhibitor zVAD-fmk. This caspase inhibitor led to an at least 5-fold improvement in cell viability without altering inhibition of protein synthesis. Interestingly, HUT-102 cells did not undergo programmed cell death after exposure to immunotoxins that kill these cells. We conclude that immunotoxins may be valuable in the treatment of cancers that are resistant toward apoptosis because their targeted killing is often facilitated by, but not completely dependent on, programmed cell death. Int. J. Cancer 87:86-94, 2000. Published 2000 Wiley-Liss, Inc.
Authors:
A Keppler-Hafkemeyer; R J Kreitman; I Pastan
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  International journal of cancer. Journal international du cancer     Volume:  87     ISSN:  0020-7136     ISO Abbreviation:  Int. J. Cancer     Publication Date:  2000 Jul 
Date Detail:
Created Date:  2000-07-13     Completed Date:  2000-07-13     Revised Date:  2007-07-24    
Medline Journal Info:
Nlm Unique ID:  0042124     Medline TA:  Int J Cancer     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  86-94     Citation Subset:  IM    
Affiliation:
Laboratory of Molecular Biology, Division of Basic Sciences, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Chloromethyl Ketones / pharmacology
Antibodies
Antigens, CD / immunology
Antigens, CD22
Antigens, Differentiation, B-Lymphocyte / immunology
Apoptosis / drug effects*
Burkitt Lymphoma / enzymology,  therapy
Caspase 3
Caspases / metabolism
Cell Adhesion Molecules*
Cell Survival / drug effects
Coumarins / pharmacology
DNA Fragmentation / drug effects
Dose-Response Relationship, Drug
Hematologic Neoplasms / therapy*
Immunoblotting
Immunotoxins / adverse effects*,  therapeutic use*
Lectins*
Leucine / metabolism
Leukemia / enzymology,  therapy
Lymphoma, T-Cell / enzymology,  therapy
Oligopeptides / pharmacology
Protease Inhibitors / pharmacology
Recombinant Proteins / metabolism
Tetrazolium Salts / pharmacology
Tumor Cells, Cultured
Chemical
Reg. No./Substance:
0/2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium; 0/Amino Acid Chloromethyl Ketones; 0/Antibodies; 0/Antigens, CD; 0/Antigens, CD22; 0/Antigens, Differentiation, B-Lymphocyte; 0/Cell Adhesion Molecules; 0/Coumarins; 0/Immunotoxins; 0/Lectins; 0/Oligopeptides; 0/Protease Inhibitors; 0/Recombinant Proteins; 0/Tetrazolium Salts; 0/antitac(FV)-PE38 recombinant immunotoxin; 0/aspartyl-glutamyl-valyl-aspartyl-7-amino-4-trifluoromethylcoumarin; 0/benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; 61-90-5/Leucine; EC 3.4.22.-/Caspase 3; EC 3.4.22.-/Caspases

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