Reduced oxygen tension results in reduced human T cell proliferation and increased intracellular oxidative damage and susceptibility to apoptosis upon activation.
Cell culture and in vitro models are the basis for much biological research, especially in human immunology. Ex vivo studies of T cell physiology employ conditions attempting to mimic the in vivo situation as closely as possible. Despite improvements in controlling the cellular milieu in vitro, most of what is known about T cell behavior in vitro is derived from experiments on T cells exposed to much higher oxygen levels than are normal in vivo. In this study, we report a reduced proliferative response and increased apoptosis susceptibility after T cell activation at 2% oxygen compared to in air. To explain this observation, we tested the hypothesis of an impaired efficacy of intracellular protective mechanisms including antioxidant levels, oxidized protein repair (methionine sulfoxide reductases), and degradation (proteasome) activities. Indeed, after activation, there was a significant accumulation of intracellular oxidized proteins at more physiological oxygen levels concomitant with a reduced GSH:GSSG ratio. Proteasome and methionine sulfoxide reductase activities were also reduced. These data may explain the increased apoptotic rate observed at more physiological oxygen levels. Altogether, this study highlights the importance of controlling oxygen levels in culture when investigating oxygen-dependent phenomena such as oxidative stress.
Larbi
Anis
A
Center for Medical Research, Tübingen Aging and Tumor Immunology Group, University of Tübingen, 72072 Tübingen, Germany. anis.larbi@medizin.uni-tuebingen.de
Cabreiro
Filipe
F
Zelba
Henning
H
Marthandan
Shiva
S
Combet
Emilie
E
Friguet
Bertrand
B
Petropoulos
Isabelle
I
Barnett
Yvonne
Y
Pawelec
Graham
G
D000975:Antioxidants / Q000378:metabolism / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D049109:Cell Proliferation; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D006801:Humans; D008213:Lymphocyte Activation; D051150:Methionine Sulfoxide Reductases / Q000378:metabolism; D008875:Middle Aged; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D010100:Oxygen / Q000378:metabolism*; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism; D012016:Reference Values; D000077185:Resveratrol; D013267:Stilbenes / Q000494:pharmacology; D013601:T-Lymphocytes / Q000166:cytology* / Q000378:metabolism*
D000975:Antioxidants; D013267:Stilbenes; D051150:Methionine Sulfoxide Reductases; D046988:Proteasome Endopeptidase Complex; D000077185:Resveratrol; D010100:Oxygen