Tumor-induced apoptosis of T cells: amplification by a mitochondrial cascade.
We have recently reported that apoptosis of T cells induced by squamous cell carcinoma of the head and neck (SCCHN) is partly Fas dependent. This tumor-induced T-cell death is mediated by the activities of caspase-8 and caspase-3 and is partially inhibited by antibodies to either Fas or Fas ligand. We report here that in contrast to apoptosis induced by agonistic anti-Fas antibody (Ab), the tumor-induced apoptotic cascade in Jurkat cells is significantly amplified by a mitochondrial loop. The involvement of mitochondria in tumor-induced apoptosis of T cells was demonstrated by changes in mitochondrial permeability transition as assessed by 3,3'-dihexiloxadicarbocyanine staining, by cleavage of cytosolic BID and its translocation to the mitochondria, by release of cytochrome c to the cytosol, and by the presence of active subunits of caspase-9 in Jurkat T cells cocultured with tumor cells. To further elucidate the significance of mitochondria in tumor-induced T-cell death, we investigated the effects of various inhibitors of the mitochondrial pathway. Specific antioxidants, as well as two inhibitors of mitochondria permeability transition, bongkrekic acid and cyclosporin A, significantly blocked the DNA degradation induced in Jurkat T cells by SCCHN cells. However, these inhibitors had no effect on cells triggered by anti-Fas Ab. Furthermore, a cell-permeable inhibitor of caspase-9, Ac-LEHD.CHO, which did not inhibit T-cell apoptosis induced by anti-Fas Ab, markedly inhibited apoptosis induced by etoposide or by coculture of Jurkat with SCCHN cells. These findings demonstrate that apoptotic cascades induced in Jurkat T lymphocytes by anti-Fas Ab or tumor cells are differentially susceptible to a panel of inhibitors of mitochondrial apoptotic events. It appears that besides the Fas-mediated pathway, additional mitochondria-dependent cascades are involved in apoptosis of tumor-associated lymphocytes. Inhibition of mitochondria-dependent cascades of caspase activation should be considered to enhance the success of immunotherapy or vaccination protocols in cancer.
Gastman
B R
BR
Department of Otolaryngology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15213, USA.
Yin
X M
XM
Johnson
D E
DE
Wieckowski
E
E
Wang
G Q
GQ
Watkins
S C
SC
Rabinowich
H
H
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology; D000975:Antioxidants / Q000494:pharmacology; D017209:Apoptosis; D051023:BH3 Interacting Domain Death Agonist Protein; D015153:Blotting, Western; D001865:Bongkrekic Acid / Q000494:pharmacology; D002232:Carbocyanines / Q000494:pharmacology; D002294:Carcinoma, Squamous Cell / Q000473:pathology; D002352:Carrier Proteins / Q000378:metabolism; D053453:Caspase 9; D020169:Caspases / Q000378:metabolism; D018920:Coculture Techniques; D016572:Cyclosporine / Q000494:pharmacology; D003574:Cytochrome c Group / Q000378:metabolism / Q000502:physiology; D003600:Cytosol / Q000378:metabolism; D004247:DNA / Q000187:drug effects; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005047:Etoposide / Q000494:pharmacology; D005455:Fluorescent Antibody Technique; D005456:Fluorescent Dyes / Q000494:pharmacology; D006258:Head and Neck Neoplasms / Q000473:pathology; D006801:Humans; D019169:Jurkat Cells; D008214:Lymphocytes / Q000473:pathology; D008856:Microscopy, Fluorescence; D008928:Mitochondria / Q000473:pathology*; D009842:Oligopeptides / Q000494:pharmacology; D021381:Protein Transport; D013347:Subcellular Fractions / Q000378:metabolism; D013601:T-Lymphocytes / Q000473:pathology*; D014407:Tumor Cells, Cultured; D019014:fas Receptor / Q000276:immunology
D000900:Anti-Bacterial Agents; D000972:Antineoplastic Agents, Phytogenic; D000975:Antioxidants; D051023:BH3 Interacting Domain Death Agonist Protein; C494814:BID protein, human; D002232:Carbocyanines; D002352:Carrier Proteins; D003574:Cytochrome c Group; D004791:Enzyme Inhibitors; D005456:Fluorescent Dyes; D009842:Oligopeptides; C420218:acetyl-leucyl-glutamyl-histidyl-aspartal; D019014:fas Receptor; D001865:Bongkrekic Acid; C007392:3,3'-dihexyl-2,2'-oxacarbocyanine; D005047:Etoposide; D016572:Cyclosporine; D004247:DNA; C506433:CASP9 protein, human; D053453:Caspase 9; D020169:Caspases