Extended Data Fig. 6: Nasal anti-CD3 modulates chronic microglial response after TBI for different treatment regimens and TBI severities. | Nature Neuroscience

Extended Data Fig. 6: Nasal anti-CD3 modulates chronic microglial response after TBI for different treatment regimens and TBI severities.

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

Extended Data Fig. 6

(a) Gating strategy for microglia. (b) Relative expression of cell types in Sham-Iso microglia (n = 3). (c) RT- qPCR of ipsilateral hemisphere 7 and 30 days post-TBI (immediate treatment males). Expression was normalized to GAPDH and presented relative to Sham-Iso. Data shown as mean ± SEM, (Sham-Iso n = 6, TBI-Iso n = 8, TBI-aCD3 n = 8) and analyzed by one-way ANOVA with Tukey’s multiple comparisons. (d) Heatmap signature of DEGs 30 days post-TBI (early treatment males) identified using DESeq2 analysis (two-sided likelihood ratio test, n = 5 mice/group, FDR-corrected P < 0.05). (e) Heatmap of genes in inflammatory response and genes of disease-associated microglia (DAM) and neurodegenerative microglia (MGnD) 30 days post-TBI (early treatment males). Genes identified with an FDR-corrected p-value < 0.05 using DESeq2 analysis are bolded (two-sided likelihood ratio test, n = 5 mice/group). (f) GSEA analysis of GO Biological Processes (BP) comparing TBI-aCD3 vs. TBI-Iso groups 30 days post-TBI (early treatment male mice). NES, normalized enrichment score. (g) RT- qPCR of ipsilateral hemisphere 30 days post-TBI (early treatment males). Expression normalized to GAPDH and presented relative to Sham-Iso. Data shown as mean ± SEM, (Sham-Iso n = 5, TBI-Iso n = 6, TBI-aCD3 n = 6) and analyzed by one-way ANOVA with Tukey’s multiple comparisons. (h) Venn diagram of DEGs in comparisons with Sham-Iso microglia group as baseline 30 days post-severe TBI (immediate treatment female mice): TBI-Iso, and TBI-aCD3. (i) Heatmap genes involved in inflammatory response and (DAM) and (MGnD) at 30 days following severe TBI (immediate treatment females). Genes identified with FDR-corrected p-value < 0.05 using DESeq2 analysis are bolded (two-sided likelihood ratio test, n = 5 mice/group). (j) RT- qPCR of ipsilateral hemisphere 30 days post-severe TBI (immediate treatment female). Expression was normalized to GAPDH and presented relative to Sham-Iso. Data shown as mean ± SEM, (Sham-Iso n = 5, TBI-Iso n = 6, TBI-aCD3 n = 6) and analyzed by one-way ANOVA with Tukey’s multiple comparisons. All data are biological replicates and are representative from two independent experiments. n.s. = non-significant.

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