Fig. 2
Local immune alterations in the brain 3 days after TBI and antibiotic induced gut microbial dysbiosis. a Experimental design. b–j Brain leucocyte profile characterization by flow cytometry. b Gating strategy of Ly6Chigh cells. Quantification of cell absolute numbers in the hippocampi, amygdalae, and cortices for c leukocytes (CD45), d CD3 T cells (CD11b−CD3+), e CD4 T cells (CD11b−CD3+CD4+), f CD8 T cells (CD11b−CD3+CD8+), g DN T cells (CD11b−CD3+CD4−CD8−), h T reg cells (CD11b−CD4+CD25+), i monocytes (CD45highCD11b+Ly6C+) and j microglia (CD45lowCD11b+). Mean values are plotted ± SEM, unpaired t test, **p < 0.01. k Representatives 20X images of ipsilateral hippocampi of sham, control-injured and antibiotic-injured mice. Scale bars are 100 µm. l Quantification of fluorescent intensity from (k), two-way ANOVA, antibiotic F(1,14) = 0.015, p = 0.9, injury F(1,14) = 29.2, p < 0.0001. m Protein cytokine and chemokine quantification from whole ipsilateral hippocampi. Mean values are plotted ± SEM, unpaired t-test, *p < 0.05. Abbreviations: VNAM, vancomycin, neomycin-sulfate, ampicillin and metronidazole; CCI, controlled cortical impact. Hip: hippocampus; Amy, amygdala; Cx, cortex