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. 2023 Jun 21;14:1181823. doi: 10.3389/fimmu.2023.1181823

Figure 3.

Figure 3

NOD2 stimulation is responsible for a mo-Macs/mo-DCs switch. (A) Experimental protocol. (B) Gating strategy for flow cytometry analysis of BMDCs upon MDP treatment. Mouse BM-derived cells were generated in vitro for 7 days in GM-CSF in the presence or not of MDP (10ug/ml). Mo-DCs were gated as CD11c+ MHCII+ CD115- CD135+ CD64+ (green) and mo-Macs were gated as CD11c+ MHCII+ CD115+ (purple). Frequencies are calculated from the CD11c+ gate. (C) Frequencies of mo-DCs and mo-Macs in the mouse BMDC culture stimulated or not with MDP at the start of the culture. (D) Mo-DCs were generated by culturing CD14+ circulating human monocytes with GM-CSF and IL-4. MDP was added at the beginning of the 5 day-culture. Morphology of the differentiating cells at day 1 and day 5 of culture in the presence or not of MDP. (E) The impact of MDP or LPS stimulation at the start of the culture on mo-Macs (CD16+CD1a-) and mo-DCs (CD16-CD1a+) differentiation was evaluated by flow cytometry at day 6, after gating on live HLA-DR+ cells. (F) mo-DCs and mo-Macs frequencies. (G) CD1a and CD16 fluorescent mean intensity (GeoMeans) (n=5). These data are representative of at least 4 independent experiments with different mice (B-C) or donors (D-G). Bars indicate mean ± SEM. Statistical significance was assessed by the non-parametric Mann-Whitney test. *P<0.05; **P<0.01.