Abstract
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.
Publication types
-
Research Support, Non-U.S. Gov't
MeSH terms
-
Antibodies, Monoclonal / pharmacology
-
CD28 Antigens / genetics
-
CD28 Antigens / immunology
-
Carmustine / pharmacology
-
Enzyme Activation / drug effects
-
Enzyme Activation / immunology
-
Genes, Reporter / drug effects
-
Genes, Reporter / immunology
-
Humans
-
Hydrogen Peroxide / pharmacology
-
I-kappa B Kinase
-
Interleukin-2 / biosynthesis
-
Interleukin-2 / genetics
-
Intracellular Fluid / metabolism*
-
Intracellular Fluid / physiology
-
JNK Mitogen-Activated Protein Kinases
-
Jurkat Cells
-
Lactic Acid / pharmacology
-
Lymphocyte Specific Protein Tyrosine Kinase p56(lck) / metabolism
-
Mitogen-Activated Protein Kinases / metabolism
-
Muromonab-CD3 / pharmacology
-
NF-kappa B / genetics
-
Oxidation-Reduction
-
Protein Serine-Threonine Kinases / metabolism
-
Protein-Tyrosine Kinases / physiology
-
Proto-Oncogene Proteins / metabolism
-
Proto-Oncogene Proteins c-fyn
-
Receptors, Antigen, T-Cell / physiology*
-
Response Elements / immunology
-
Signal Transduction / immunology*
-
Sulfhydryl Compounds / metabolism*
-
Sulfhydryl Compounds / physiology
-
T-Lymphocytes / enzymology
-
T-Lymphocytes / metabolism
-
Transcription, Genetic / drug effects
-
Transcription, Genetic / immunology
Substances
-
Antibodies, Monoclonal
-
CD28 Antigens
-
Interleukin-2
-
Muromonab-CD3
-
NF-kappa B
-
Proto-Oncogene Proteins
-
Receptors, Antigen, T-Cell
-
Sulfhydryl Compounds
-
Lactic Acid
-
Hydrogen Peroxide
-
Protein-Tyrosine Kinases
-
FYN protein, human
-
Lymphocyte Specific Protein Tyrosine Kinase p56(lck)
-
Proto-Oncogene Proteins c-fyn
-
Protein Serine-Threonine Kinases
-
CHUK protein, human
-
I-kappa B Kinase
-
IKBKB protein, human
-
IKBKE protein, human
-
JNK Mitogen-Activated Protein Kinases
-
Mitogen-Activated Protein Kinases
-
Carmustine