Journal of Biochemistry Advance Access originally published online on December 13, 2006
Journal of Biochemistry 2007 141(2):157-172; doi:10.1093/jb/mvm019
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© 2006 The Japanese Biochemical Society.
Characterization of Galectin-9-Induced Death of Jurkat T Cells
1Department of Endocrinology, Faculty of Medicine, Kagawa University, Kagawa, Japan; 2Department of Cell Regulation, Faculty of Medicine, Kagawa University, Kagawa, Japan; 3Department of Immunology and Immunopathology, Faculty of Medicine, Kagawa University, Kagawa, Japan; and 4Galpharma Co., Inc., Kagawa, Japan
*To whom correspondence should be addressed. Tel: +81-87-891-2106, Fax: +81-87-891-2108, E-mail: tnaka{at}med.kagawa-u.ac.jp
Received October 18, 2006; Accepted November 22, 2006
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Abstract |
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Galectin-9, a mammalian lectin with affinity for ß-galactosides, is known as an apoptosis inducer of activated T lymphocytes. In the present study, we examined the properties of galectin-9-mediated cell death of Jurkat T cells. Galectin-9NC (wild-type), consisting of two CRDs (N-terminal and C-terminal carbohydrate recognition domains), and derivatives of it, galectins-9-NN and -9-CC, induced Jurkat T-cell apoptosis. However, a single CRD (galectin-9NT or -CT) had no effect, suggesting the stable dimeric structure of two CRDs is required for the activity. The apoptosis was inhibited by pretreatment with an N-glycan synthesis inhibitor, indicating that the expression of N-glycans in the cells is essential for galectin-9-induced apoptosis. We previously showed that the apoptosis of MOLT-4 cell is mediated by galectin-9 via a Ca2+-calpain-caspase-1-dependent pathway. In Jurkat cells, the cell death by galectin-9, was insufficiently suppressed by caspase inhibitors, Ca2+-chelator or calpain inhibitor. Furthermore, we observed the loss of mitochondrial membrane potential and significant AIF release in galectin-9-treated cells. These findings suggest that caspase-dependent and-independent death pathways exist in Jurkat cells, and the main pathway might vary with the T-cell type.
Key Words: galectin-9, galectin, apoptosis, Jurkat, T cell
Abbreviations:
7-AAD, 7-amino-actinomycin D; ActD, actinomycin D; AIF, apoptosis-inducing factor; BAPTA, O,O'-bis(2-aminophenyl)ethyleneglycol-N,N,N',N'-tetraacetic acid; CCCP, carbonyl cyanide 3-chlorophenylhydrazone; CRD, carbohydrate recognition domain; DiOC2(3), 3,3'-diethyloxacarbocyanine iodide; DMEM, Dulbecco's Modified Eagle's Medium; DMNJ, deoxymannojirimycin; EDTA, ethylenediamine-tetraacetic acid; EGTA, ethyleneglycol-tetraacetic acid; EndoG, endonuclease G; Fluo 3, 1-[2-amino-5-(2,7-dichloro-6-hydroxy-3-oxo-9-xanthenyl)phenoxy]-2-(2-amino-5-methylphenoxy)ethane-N,N,N',N'-tetraacetic acid; Gal, galectin; 
m, mitochondrial membrane potential; mAb, monoclonal antibody; PE, phycoerythin; PS, phosphatidylserine; RCA-120, Ricinus communis agglutinin; TUNEL, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labelling
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