Fig. 1: IL-2 and HS colocalize at sites of autoimmune neuroinflammation.
From: Regulatory T cells use heparanase to access IL-2 bound to extracellular matrix in inflamed tissue
a Imaris surface rendering of immunofluorescent staining for IL-2 (green), HS (magenta) and CD45 (yellow) in naïve and EAE spinal cord tissue (29 days post immunization (dpi)). DAPI nuclear counterstain shown in blue. Representative of 3 separate EAE experiments. b Detail of IL-2 (green), HS (magenta) and CD45 (yellow) immunoreactivity in an EAE lesion. c Imaris quantification of abundance of IL-2 and HS immunoreactivity, and fraction of IL-2 colocalized with HS in naïve and EAE spinal cord tissue at different time points (early acute: 14 dpi, late acute: 29 dpi, chronic 40 dpi). Imaris colocalization analysis was done in areas surrounding CD45 cell infiltration (perilesion), areas with CD45 cell infiltration (lesion) and in the glia limitans underlying the meninges (pia). Shown are mean + SEM, n = 3 (naïve), 5 (early acute), 4 (late acute and chronic) separate areas analyzed. P values determined by unpaired two-tailed Welch’s t test with Benjamini, Krieger and Yekutieli correction for multiple comparisons. d Immunofluorescent staining for IL-2 (green) and HS (magenta) of a cerebellar EAE lesion that was treated with heparinase from Flavobacterium heparinum before immunofluorescent staining, or buffer treatment of a serial section as a control. CD45 (blue) staining is shown to depict areas with immune cell infiltration. Representative of 2 separate experiments. e Imaris quantification of IL-2 colocalized with HS before (red) and after heparinase (gray) treatment. Shown are mean + SEM, n = 3 separate areas analyzed. P values determined by two-way ANOVA with Sidak’s multiple-comparison correction.
