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Loss of Nrf2 in bone marrow-derived macrophages impairs antigen-driven CD8+ T cell function by limiting GSH and Cys availability

 
: Sha, L.K.; Sha, W.; Kuchler, L.; Daiber, A.; Giegerich, A.K.; Weigert, A.; Knape, T.; Snodgrass, R.; Schröder, K.; Brandes, R.P.; Brüne, B.; Knethen, A. von

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Free radical biology and medicine 83 (2015), pp.77-88
ISSN: 0891-5849
ISSN: 1873-4596
English
Journal Article
Fraunhofer IME ()

Abstract
NF-E2-related factor 2 (Nrf2), known to protect against reactive oxygen species, has recently been reported to resolve acute inflammatory responses in activated macrophages. Consequently, disruption of Nrf2 promotes a proinflammatory macrophage phenotype. In the current study, we addressed the impact of this macrophage phenotype on CD8+ T cell activation by using an antigen-driven coculture model consisting of Nrf2-/- and Nrf2+/+ bone marrow-derived macrophages (BMDM) and transgenic OT-1 CD8+ T cells. OT-1 CD8+ T cells encode a T cell receptor that specifically recognizes MHC class I-presented ovalbumin OVA(257-264) peptide, thereby causing a downstream T cell activation. Interestingly, coculture of OVA(257-264)-pulsed Nrf2-/- BMDM with transgenic OT-1 CD8+ T cells attenuated CD8+ T cell activation, proliferation, and cytotoxic function. Since the provision of low-molecular-weight thiols such as glutathione (GSH) or cysteine (Cys) by macrophages limits antigen-driven CD 8+ T cell activation, we quantified the amounts of intracellular and extracellular GSH and Cys in both cocultures. Indeed, GSH levels were strongly decreased in Nrf2-/- cocultures compared to wild-type counterparts. Supplementation of thiols in Nrf2-/- cocultures via addition of glutathione ester, N-acetylcysteine, -mercaptoethanol, or cysteine itself restored T cell proliferation as well as cytotoxicity by increasing intracellular GSH. Mechanistically, we identified two potential Nrf2-regulated genes involved in thiol synthesis in BMDM: the cystine transporter subunit xCT and the modulatory subunit of the GSH-synthesizing enzyme -GCS (GCLM). Pharmacological inhibition of -GCS-dependent GSH synthesis as well as knockdown of the cystine antiporter xCT in Nrf2+/+ BMDM mimicked the effect of Nrf2-/- BMDM on CD8+ T cell function. Our findings demonstrate that reduced levels of GCLM as well as xCT in Nrf2-/- BMDM limit GSH availability, thereby inhibiting antigen-induced CD8+ T cell function.

: http://publica.fraunhofer.de/documents/N-351994.html