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Galectin-1 induced activation of the mitochondrial apoptotic pathway: evidence for a connection between death-receptor and mitochondrial pathways in human Jurkat T lymphocytes

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Abstract

Galectin-1 (gal-1) triggers T cell death by several distinct intracellular pathways including the activation of the death-receptor pathway. The aim of this study was to investigate whether gal-1 induced activation of the death-receptor pathway in Jurkat T lymphocytes mediates apoptosis via the mitochondrial pathway linked by truncated Bid (tBid). We demonstrate that gal-1 induced proteolytic cleavage of the death agonist Bid, a member of the Bcl-2/Bcl-xL family and a substrate of activated caspase-8, was inhibited by caspase-8 inhibitor II (Z-IETD-FMK). Downstream of Bid, gal-1 stimulated mitochondrial cytochrome c release as well as the activation and proteolytic processing of initiator procaspase-9 were effectively decreased by caspase-8 inhibitor II. Blocking of gal-1 induced cleavage of effector procaspase-3 by caspase-8 inhibitor II as well as by caspase-9 inhibitors I (Z-LEHD-FMK) and III (Ac-LEHD-CMK) indicates that receptor and mitochondrial pathways converged in procaspase-3 activation and contribute to proteolytic processing of effector procaspase-6 and -7. Western blot analyses and immunofluorescence staining revealed that exposure of Jurkat T cells to gal-1 resulted in the cleavage of the DNA-repair enzyme poly (ADP-ribose) polymerase, cytoskeletal α-fodrin, and nuclear lamin A as substrates of activated caspases. Our data demonstrate that Bid provides a connection between the death receptor and the mitochondrial pathway of gal-1 induced apoptosis in human Jurkat T lymphocytes.

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Abbreviations

Bp:

Base pair

BSA:

Bovine serum albumin

CD:

Cluster of differentiation

CHAPS:

3-[(3-Cholamidopropyl)dimethylammonio]-1-propanesulfonate

CRD:

Carbohydrate recognition domain

DTT:

Dithiothreitol

ECL:

Enhanced chemiluminescence

EDTA:

Ethylene-diaminetetraacetic acid

EGTA:

Ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid

FCS:

Fetal calf serum

Gal-1:

Galectin-1

HEPES:

N-2-hydroxyethyl-piperazine-N-2-ethanesulfonic acid

IgG:

Immunoglobulin G

HRP:

Horseradish peroxidase

kDa:

kilodalton

mAb:

Monoclonal antibody

NP-40:

Nonidet P-40

pAb:

Polyclonal antibody

PIPES:

Piperazine-N,N′-bis(2-ethanesulfonic) acid

PAGE:

Polyacrylamide gel electrophoresis

PARP:

Poly (ADP-ribose) polymerase

PBS:

Phosphate-buffered saline

PI:

Propidium iodide

SDS:

Sodium dodecyl sulphate

TBS:

Tris-buffered saline

TCR:

T cell receptor

Tris:

Tris(hydroxymethyl) aminomethane

T:

Tween 20

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Acknowledgments

This work was supported by a grant from the Deutsche Forschungsgemeinschaft (DFG, WA 1771/1-1). We gratefully acknowledge the excellent technical assistance of Mrs. G. Gaede.

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Authors and Affiliations

  1. Medical Faculty, Institute of Medical Biochemistry and Molecular Biology, Schillingallee 70, Rostock, Germany

    Falko Lange, Bettina Brandt, Markus Tiedge & Hermann Walzel

  2. Department of Pathology, Electron Microscopic Center and Branch of Medical Biology, University of Rostock, Strempelstrasse 14, 18057, Rostock, Germany

    Ludwig Jonas

  3. First Department of Obstetrics and Gynaecology, Ludwig Maximilians University of Munich, Maistrasse 11, 80337, Munich, Germany

    Udo Jeschke

  4. Unit of Reproductive Biology, Research Institute of the Biology of Farm Animals, Wilhelm-Stahl-Allee 2, 18196, Dummerstorf, Germany

    Ralf Pöhland

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  1. Falko Lange
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  2. Bettina Brandt
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Correspondence to Hermann Walzel.

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Lange, F., Brandt, B., Tiedge, M. et al. Galectin-1 induced activation of the mitochondrial apoptotic pathway: evidence for a connection between death-receptor and mitochondrial pathways in human Jurkat T lymphocytes. Histochem Cell Biol 132, 211–223 (2009). https://doi.org/10.1007/s00418-009-0597-x

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