Multiple Sclerosis (MS) is a demyelinating/inflammatory disease of the central nervous system. Relapsing‐remitting MS is characterized by a relapsing phase with clinical symptoms and the production of inflammatory cell infiltrates, and a period of remission during which patients recover partially. Myeloid‐derived suppressor cells (MDSCs) are immature cells capable of suppressing the inflammatory response through Arginase‐I (Arg‐I) activity, among other mechanisms. Here, we have identified Arg‐I+‐MDSCs in the spinal cord during experimental autoimmune encephalomyelitis (EAE), cells that were largely restricted to the demyelinating plaque and that always exhibited the characteristic MDSC surface markers Arg‐I/CD11b/Gr‐1/M‐CSF1R. The presence and density of Arg‐I+‐cells, and the proportion of apoptotic but not proliferative T cells, were correlated with the EAE time course: peaked in parallel with the clinical score, decreased significantly during the remitting phase and completely disappeared during the chronic phase. Spinal cord‐isolated MDSCs of EAE animals augmented the cell death when co‐cultured with stimulated control splenic CD3 T cells. These data point to an important role for MDSCs in limiting inflammatory damage in MS, favoring the relative recovery in the remitting phase of the disease. Thus, the MDSC population should be considered as a potential therapeutic target to accelerate the recovery of MS patients.

We have previously described that IFN-γ induces cyclooxygenase 2 and inducible NO synthase expression by a mechanism that involved endogenously produced TNF-α. In this study, we report that TNF-α production is induced by IFN-γ treatment in the murine macrophage cell line RAW 264.7. TNF-α mRNA levels are increased in cells treated with IFN-γ in a time-dependent manner and IFN-γ also increased human TNF-α promoter-dependent transcription. Two regions in the TNF-α promoter seem to be responsible for the IFN-γ response: a distal region between −1311 and −615 bp of the human TNF-α promoter, and a proximal region located between −95 and −36 bp upstream of the transcriptional start. In contrast, IFN-γ stimulation induces the expression of the transcription factors IRF-1 and IRF-8. Overexpression of these transcription factors produces an increase in the transcriptional activity of the human TNF-α promoter. There is a correlation between the regions of the TNF-α promoter responsible of the transcriptional activation elicited by IRF-1 and IRF-8 and those required for IFN-γ response. In addition, IRF-1 and IRF-8 are recruited to the TNF-α promoter in IFN-γ-treated RAW 264.7 cells, as demonstrated by chromatin immunoprecipitation assays. Moreover, overexpression of IRF-1 and IRF-8 induces TNF-α production in unstimulated RAW 264.7 macrophages, comparable to the production of TNF-α elicited by IFN-γ stimulation, and silencing of IRF-1 and/or IRF-8 with specific small interfering RNAs, decreases IFN-γ-elicited TNF-α production. In summary, IFN-γ treatment induces TNF-α expression at transcriptional level requiring the coordinate action of IRF-1 and IRF-8.

IFN-gamma induces cyclooxygenase (COX)-2 expression and PG production in mouse macrophage cells. IFN-gamma activates COX-2 promoter-driven transcription. Deletion of the IFN sequence regulatory element (ISRE) I -1541/-1522 and ISRE II -1215/-1206 sites of the mouse COX-2 promoter minimally decrease this IFN-gamma induction. In contrast, deletion of the -965/-150 region from the COX-2 promoter abrogated IFN-gamma induction. In this region a NF-kappaB site has been described and mutation of this site impairs the induction of the full COX-2 promoter by IFN-gamma. Moreover, IFN-gamma induction of the COX-2 promoter was also strongly reduced by transfection of plasmid encoding the NF-kappaB inhibitor, IkappaBalpha. Interestingly, IFN-gamma induction of the COX-2 and PGE(2) synthesis was absent in macrophages from TNF(-/-) mice, and neutralizing anti-TNF Abs inhibited COX-2 promoter induction by IFN-gamma in RAW 264.7 macrophages. Moreover, NF-kappaB activity was induced late after stimulation with IFN-gamma correlating with the effect of autocrine TNF, and this NF-kappaB activation was absent in macrophages from TNF(-/-) mice. Taken together our results suggest a model in which IFN-gamma-induced TNF activates NF-kappaB, which is required for full COX-2 expression.