Fig. 4: Nasal aCD3 increased microglial phagocytic capacity after TBI in an IL-10-dependent manner. | Nature Neuroscience

Fig. 4: Nasal aCD3 increased microglial phagocytic capacity after TBI in an IL-10-dependent manner.

From: Nasal anti-CD3 monoclonal antibody ameliorates traumatic brain injury, enhances microglial phagocytosis and reduces neuroinflammation via IL-10-dependent Treg–microglia crosstalk

Fig. 4

a, Heatmap of microglial phagocytosis genes 7 d after TBI. Genes identified with FDR-corrected P < 0.05 using DESeq2 analysis are indicated by an asterisk (two-sided LRT, n = 4 mice per group). Genes were identified from the GOBP term phagocytosis and microglial phagocytosis genes51,61. b, Schematic presenting a phagocytosis functional study (created with BioRender.com). c, Immunofluorescence of lesion (7 d post-TBI) for apoptotic neurons (blue) and P2ry12 (red) showing engulfment of apoptotic neurons by P2ry12. Scale bars, 100 μm and 50 μm for the enlarged image. d, Phagocytosis experiment where mice were injected with labeled apoptotic neurons and sacrificed 4 h post-injection. The gating strategy shows phagocytic positive microglia and data are shown as box plots (min., max., IQR, median) and n = 5 mice per group were used. Data were analyzed by two-sided, unpaired Student’s t-test. e, Clustered heatmap of DEGs of aggregated samples for phagocytic (+P) and nonphagocytic (−P) microglia 7 d post-TBI and 4 h post-injection of apoptotic neurons identified using DESeq2 analysis (two-sided LRT, n = 3-4 mice per group, FDR-corrected P < 0.05). f, Bar plots with log2(fold-changes) of genes from e pertinent to microglial phagocytosis and related functions in the following comparisons: TBI-iso (+P) versus TBI-iso (−P) and TBI-aCD3 (+P) versus TBI-Iso (−P). g, GSEA analysis of GOBP 7 d post-TBI and 4 h post-injection of apoptotic neurons based on pairwise comparisons: TBI-iso (+P) versus TBI-Iso (−P), TBI-aCD3 (+P) versus TBI-Iso (−P) and TBI-aCD3 (−P) versus TBI-Iso (−P). The asterisk indicates enriched terms (q-value < 0.05). h, Selected top canonical pathways from IPA analysis of DEGs in phagocytic TBI-aCD3 microglia compared with phagocytic and nonphagocytic TBI-iso microglial groups at 7 d post-TBI and 4 h post-injection of apoptotic neurons. i, Predicted upstream regulator in TBI-aCD3 (+P) versus TBI-iso (−P). j, Phagocytosis experiment with similar design to b. Data are shown as box plots (min., max., IQR, median) and n = 5 mice per group were used. The data were analyzed by one-way ANOVA with Tukey’s multiple comparisons. All data are biological replicates and represent two independent experiments.

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