Fig. 5: MAIT cells and macrophages crosstalk induces an inflammatory response in vitro. | Nature Communications

Fig. 5: MAIT cells and macrophages crosstalk induces an inflammatory response in vitro.

From: Mucosal-associated invariant T cells promote inflammation and intestinal dysbiosis leading to metabolic dysfunction during obesity

Fig. 5

a, b Wild-type (clear background) or MR1−/− (gray background) M0 macrophages were co-cultured for 72 h with spleen MAIT cells, either naive (nMAIT) or pre-activated (acMAIT). a Data showing the frequency of M1 macrophages (CD206-CD11c+) and M2 macrophages (CD206+CD11c−) among CD11b+F4/80+ total macrophages and M1/M2 ratio (n = 12 samples over two independent experiments). b Wild-type or MR1−/− M0 macrophages were co-cultured for 72 h with spleen pre-activated MAIT cells (acMAIT) in presence of cytokine-neutralizing antibodies (anti-IL-17, anti-IFNγ, and/or anti-TNF). Data show the frequency of M1 macrophages (CD206−CD11c+) and M2 macrophages (CD206+CD11c−) among CD11b+F4/80+ total macrophages and M1/M2 ratio (n = 8 samples over two independent experiments). c Relative expression of Mr1 in M0, M2, or M1 macrophages and surface staining for MR1 in in vitro-differentiated macrophages. Culture media contained 100 μM of Ac-6-FP. d–e Spleen nMAIT cells were co-cultured with M0-, M2-, or M1-differentiated macrophages for 48 h in presence of 5-OP-RU agonist molecule. c Data showing the frequency of M1 macrophages (CD206−CD11c+) and M2 macrophages (CD206+CD11c−) among CD11b+F4/80+ total macrophages co-cultured with acMAIT cells in medium with or without anti-IL-17A, anti-IFNγ and/or anti-TNFα neutralizing antibodies (n = 4 samples). d CD44 and CD69 surface marker expression on nMAIT cells and intracellular staining of nMAIT cells for TNFα, IFNγ, and IL-17A (n = 20 samples from four independent experiments). e Intracellular staining of MAIT cells co-cultured either with WT M0, M2, or M1 macrophages (round symbol, clear background) or MR1−/− M0, M2, or M1 macrophages (diamond symbol, gray background) for TNF-α, IFN-γ, and IL-17A (n = 7 samples). Data are representative of at least three independent experiments. For a and a–e, each symbol represents an individual mouse (small horizontal lines indicate the mean ± S.E.M.). In b, in M1/M2 ratio graph, $ symbol represents statistics between WT macrophages vs. MR1−/− macrophages cultured in the same conditions. All statistical analyses were performed by two-tailed Mann–Whitney test. *P < 0.05, **P < 0.01, ***P < 0.001 (see also Supplementary Fig. 11).

Back to article page