Figure 2 B-cell very late antigen-4 (VLA-4) deficiency reduced CNS accumulation of B cells, but not proinflammatory or regulatory T cells (Treg), in myelin.

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Figure 2 B-cell very late antigen-4 (VLA-4) deficiency reduced CNS accumulation of B cells, but not proinflammatory or regulatory T cells (Treg), in myelin oligodendrocyte glycoprotein (MOG) p35-55-induced experimental autoimmune encephalomyelitis (EAE)‏ B-cell very late antigen-4 (VLA-4) deficiency reduced CNS accumulation of B cells, but not proinflammatory or regulatory T cells (Treg), in myelin oligodendrocyte glycoprotein (MOG) p35-55-induced experimental autoimmune encephalomyelitis (EAE) CD19cre/α4f/f or control mice were immunized with MOG p35-55. Analyses were performed 14 days after immunization. (A) CNS-infiltrating CD19+ B cells (gated on viable CD45hi lymphocytes) were collected at peak disease. Flow cytometry plots show 1 representative mouse each (n = 5–6 mice/group; pooled from 2 independent experiments). **p ≤ 0.01; Mann-Whitney U test. (B) Immunohistochemistry for T (CD3+) and B (B220+) cells in CNS infiltrates. B cells were not detected in spinal cord meningeal and inflammatory foci in CD19cre/α4f/f mice with EAE (upper left panel), whereas B cells were detected in CD19cre control mice (upper right panel, arrows). There were comparable T-cell infiltrates in inflammatory foci in corresponding adjacent serial sections (lower panels). Each panel shows a spinal nerve root. Scale bar, 30 μm, applies to the 4 panels. Semiquantitative analysis of n = 6 CD19cre/α4f/f mice and 3 control mice (right). **p ≤ 0.01; ns = not significant; unpaired t test. (C, D) CNS-infiltrating Th1 (interferon [IFN]–γ-secreting) and Th17 (interleukin [IL]–17A-secreting) cells (C) and CD25+FoxP3+ Treg (D) were measured by intracellular cytokine staining (ICS). Flow cytometry plots (gated on viable CD4+ T cells) show 1 representative mouse per group (n = 6 mice/group; pooled from 2 independent experiments). Data shown in graphs represent mean frequency (%) ± SEM. ns = not significant; unpaired t test. Klaus Lehmann-Horn et al. Neurol Neuroimmunol Neuroinflamm 2016;3:e212 © 2016 American Academy of Neurology