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Fig. 3 | Journal of Neuroinflammation

Fig. 3

From: A GM-CSF-neuroantigen tolerogenic vaccine elicits inefficient antigen recognition events below the CD40L triggering threshold to expand CD4+ CD25+ FOXP3+ Tregs that inhibit experimental autoimmune encephalomyelitis (EAE)

Fig. 3

Administration of a CD40 agonist in vivo inhibited the Treg-inductive activity of GMCSF-MOG. On day − 2 and 0, 2D2-FIG mice (n = 7/group) were injected i.p. with 100 μg of an anti-CD40 mAb (clone FGK4.5, rat anti-mouse CD40, IgG2a) or control mAb (clone 2A2, rat anti-trinitrophenol, IgG2a) in 500 μl saline. All mice were injected with 4 nmol of GMCSF-MOG on day 0. PBMC were analyzed on day − 8 before vaccination and day 3 post-vaccination for side-scatter (SSC), CD3, and FOXP3. Lymph nodes were harvested and analyzed on day 4 for CD4, CD11b, MHCII, and FOXP3. Shown are a representative histograms analyzed for MHCII expression (x-axis), b MHCII median fluorescence intensity, and c numbers of CD4− CD11b− MHCII+ cells (B cells) and CD11b+ CD4− cells (myeloid APC) from lymph nodes on day 4. Also shown are d representative dot plots of CD3+ T cells analyzed for SSC (y-axis) and FOXP3 (x-axis) from blood on day 3 together with percentages e and numbers f of Tregs (per μl of blood) on days − 8 and 3. Statistical significance was analyzed by use of a one-tailed t test. (*p < 0.05, ***p < 0.001). These data are representative of two independent experiments. Error bars represent SEM

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Journal of Neuroinflammation

ISSN: 1742-2094