Enhancement of T cell receptor signaling by a mild oxidative shift in the intracellular thiol pool.
Exposure of T cells to the macrophage products hydrogen peroxide (HP) or L-lactate (LAC) was previously shown to enhance IL-2 production and to modulate glutathione (GSH) status. We now found that 50 microM HP and 30 mM LAC enhanced strongly the transcription from the IL-2 promoter in Jurkat T cells after stimulation with anti-CD28 together with or without anti-CD3 but not with anti-CD3 Abs alone. Therefore, we used anti-CD3 plus anti-CD28-stimulated cells to investigate the effect of the GSH reductase inhibitor 1, 3-bis(2-chloroethyl)-1-nitrosourea (BCNU) on the signal cascade. BCNU enhanced the transcription to a similar extent as HP or LAC. Lowering the intracellular GSH/GSH disulfide ratio by BCNU, HP, or NO resulted in all cases in the fulminant enhancement of Jun-N-terminal kinase and p38 mitogen-activated protein kinase but not extracellular signal-regulated kinase 1/2. Jun-N-terminal kinase and NF-kappaB activation was enhanced through pathways involving Rac, Vav1, PKCTheta, p56(lck), p59(fyn), and IkappaB kinases. In a cell-free system, the autophosphorylation of rFyn was stimulated by GSH disulfide but not by HP. These findings suggest that the oxidation of the cellular thiol pool may play a role as an amplifying mechanism for TCR/CD3 signals in immune responses.
Hehner
S P
SP
Department of Immunochemistry, German Cancer Research Center (DKFZ), Heidelberg, Germany.
Breitkreutz
R
R
Shubinsky
G
G
Unsoeld
H
H
Schulze-Osthoff
K
K
Schmitz
M L
ML
Dröge
W
W
D000911:Antibodies, Monoclonal / Q000494:pharmacology; D018106:CD28 Antigens / Q000235:genetics / Q000276:immunology; D002330:Carmustine / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects / Q000276:immunology; D017930:Genes, Reporter / Q000187:drug effects / Q000276:immunology; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D051550:I-kappa B Kinase; D007376:Interleukin-2 / Q000096:biosynthesis / Q000235:genetics; D007424:Intracellular Fluid / Q000378:metabolism* / Q000502:physiology; D048031:JNK Mitogen-Activated Protein Kinases; D019169:Jurkat Cells; D019344:Lactic Acid / Q000494:pharmacology; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck) / Q000378:metabolism; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D016853:Muromonab-CD3 / Q000494:pharmacology; D016328:NF-kappa B / Q000235:genetics; D010084:Oxidation-Reduction; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000502:physiology; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D051076:Proto-Oncogene Proteins c-fyn; D011948:Receptors, Antigen, T-Cell / Q000502:physiology*; D020218:Response Elements / Q000276:immunology; D015398:Signal Transduction / Q000276:immunology*; D013438:Sulfhydryl Compounds / Q000378:metabolism* / Q000502:physiology; D013601:T-Lymphocytes / Q000201:enzymology / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects / Q000276:immunology
D000911:Antibodies, Monoclonal; D018106:CD28 Antigens; D007376:Interleukin-2; D016853:Muromonab-CD3; D016328:NF-kappa B; D011518:Proto-Oncogene Proteins; D011948:Receptors, Antigen, T-Cell; D013438:Sulfhydryl Compounds; D019344:Lactic Acid; D006861:Hydrogen Peroxide; D011505:Protein-Tyrosine Kinases; C494957:FYN protein, human; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck); D051076:Proto-Oncogene Proteins c-fyn; D017346:Protein Serine-Threonine Kinases; C496559:CHUK protein, human; D051550:I-kappa B Kinase; C496560:IKBKB protein, human; C496565:IKBKE protein, human; D048031:JNK Mitogen-Activated Protein Kinases; D020928:Mitogen-Activated Protein Kinases; D002330:Carmustine