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. 2022 Jul 14;13:4074. doi: 10.1038/s41467-022-31723-4

Fig. 4. Clec2d deficiency impairs MDSC activation to alleviate C. neoformans infection.

Fig. 4

a 138 downregulated (green) and 314 upregulated genes (red) in BM-derived MDSCs from wild-type (WT) or Clec2d−/− mice, which were generated by GM-CSF (GM, 40 ng/ml) + GXM (10 μg/well) for 5 days and then treated with GXM (10 μg/well) for 3 h. The volcano plot showed differentially expressed genes based on absolute fold change >2 and FDR <0.05. b Heatmap of differentially expressed MDSC-related and pro-inflammatory genes as shown in a. c Schematic strategy for generating wild-type and Clec2d-deficient BM-derived MDSCs (MDSCWT and MDSCClec2d−/−), which were co-cultured with T-cells or adoptively transferred to wild-type or Clec2d-deficient mice (1 × 106 cells/mouse). d mRNA levels of Arg1 and Nos2 in wild-type and Clec2d-deficient BM-derived MDSCs, which were generated with GM-CSF (GM, 40 ng/ml, MDSCN), GM + IL-6 (40 ng/ml, MDSCIL-6), or GM + GXM (10 μg/well, MDSCGXM) for 5 days. e Proliferation frequency of CD4+ or CD8+ T-cells, which were co-cultured for 72 h with MDSCN, MDSCIL-6, or MDSCGXM as shown in d. f, g Fungal burden (f) or mRNA levels of Arg1 and Nos2 (g) in lungs of Cap59-infected mice (1 × 105 CFU/mouse) on day 14, which are adoptively transferred with MDSCN, MDSCIL-6, or MDSCGXM (1 × 106 cells/mouse) as shown in d. h–k Fungal burden (h), representative histological images (i), mRNA levels of Arg1 and Nos2 (j), or the frequency of IFNγ+ Th1 and IL-17A+ Th17 (k) in lungs of C. neoformans strain H99-infected Clec2d-/- mice (1 × 103 CFU/mouse) on Day 14, which are adoptively transferred with MDSCWT and MDSCClec2d-/- (1 × 106 cells/mouse) generated by GM (40 ng/ml) + GXM (10 μg/well). Scale bars = 2000 μm, arrows indicate yeasT-cells in granulomatous lesions. Data were presented as mean ± SEM, n = 3 (d–h, j, k) biologically independent samples. Data were analyzed by unpaired two-sided Student’s t-test in e, f–h, j, k and one-way ANOVA adjusted for multiple comparisons in d. Source data are provided as a Source Data file.