Diminished activation of the MAP kinase pathway in CD3-stimulated T lymphocytes from old mice.
Stimulation of the ERK family of protein kinases ('extracellular signal regulated kinases', also known as MAP kinases) plays an important role in the activation of many cell types, including T lymphocytes. ERKs are activated when they are phosphorylated by an upstream activator, the dual-specific protein kinase MEK. To see if aging leads to an impairment of MEK activation in mouse T cells, we used a mobility shift assay in which activation of MEK leads to phosphorylation and altered mobility of ERK-2 kinase. Similarly, we monitored mobility of pp90rsk, a known ERK substrate, as an indication of ERK function. We found an age-related decline in the ability of mouse T cells to activate both MEK and ERK function in response to stimulation by antibodies to the CD3 chain of the T cell receptor. Aging did not alter the kinetics of enzyme activation, but did diminish (by about 2-fold) the maximal level of substrate converted into the slower migrating form. Naive and memory CD4 T cells from young mice were equally able to convert ERK2 to its slower migrating form, suggesting that the decline in MEK function is not likely to be attributable to the shift, with age, from naive to memory T cell predominance. Our data suggest that age-dependent declines in gene activation, including genes for key cytokines like IL-2, may be due to declines in the upstream signals that lead to activation of the MEK/ERK protein kinase cascade.
Gorgas
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Department of Pathology, University of Michigan, School of Medicine, Ann Arbor 48109, USA.
Butch
E R
ER
Guan
K L
KL
Miller
R A
RA
D000375:Aging; D000818:Animals; D000906:Antibodies / Q000276:immunology; D017252:CD3 Complex / Q000276:immunology*; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000276:immunology*; D003432:Cross-Linking Reagents; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D008297:Male; D051379:Mice; D019950:Mitogen-Activated Protein Kinase 1; D008934:Mitogens / Q000494:pharmacology; D010835:Phytohemagglutinins / Q000494:pharmacology; D017346:Protein Serine-Threonine Kinases / Q000276:immunology; D011505:Protein-Tyrosine Kinases / Q000276:immunology; D019893:Ribosomal Protein S6 Kinases; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology*
D000906:Antibodies; D017252:CD3 Complex; D003432:Cross-Linking Reagents; D008934:Mitogens; D010835:Phytohemagglutinins; D011505:Protein-Tyrosine Kinases; D017346:Protein Serine-Threonine Kinases; D019893:Ribosomal Protein S6 Kinases; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D019950:Mitogen-Activated Protein Kinase 1