Journal: Mediators of Inflammation

Article Title: Exaggerated IL-15 and Altered Expression of foxp3+ Cell-Derived Cytokines Contribute to Enhanced Colitis in Nlrp3−/− Mice

doi: 10.1155/2016/5637685

Figure Lengend Snippet: Nlrp3−/− mice exhibit increased numbers of IL-15 expressing DCs in the MLN and colonic following a 7-day course of DSS. (a) The quantification of IL-15 in colonic tissue isolated from WT and Nlrp3−/− on day 7 of DSS. ∗ denote p < 0.05 compared to WT; n = 5/group. Representative images of (b) WT and (c) Nlrp3−/− colonic sections stained for IL-15; scale bar = 100 microns. Flow cytometric analysis of IL-15 expression on the surface of all DCs isolated from the MLNs of WT and Nlrp3−/− on day 7 of DSS, expressed as a percentage of IL-15 expressing cells (d) and mean fluorescence intensity (MFI) per cell (e); ∗ denotes p < 0.05 compared to WT; n = 6/group. Flow cytometric analysis of IL-15 expression on the surface of lymphoid (CD11c+/CD8+; LDC) and plasmacytoid (CD11b+/B220+/CD11c−; pDC) DCs isolated from the LP of WT and Nlrp3−/− mice on day 7 of DSS, expressed as a percentage of IL-15 expressing cells (f) and MFI per cell (g); ∗ denotes p < 0.05 compared to WT; n = 5/group.

Article Snippet: Sections were incubated with anti-mouse foxp3 (BioLegend Cat.# 320001; San Diego, CA, USA) or biotin-conjugated anti-mouse IL-15 (R&D Systems Cat.# BAF 447; Burlington, ON, CAN).

Techniques: Expressing, Isolation, Staining, Fluorescence

Journal: Mediators of Inflammation

Article Title: Exaggerated IL-15 and Altered Expression of foxp3+ Cell-Derived Cytokines Contribute to Enhanced Colitis in Nlrp3−/− Mice

doi: 10.1155/2016/5637685

Figure Lengend Snippet: BM-derived DCs from Nlrp3−/− mice exhibit increased frequency of IL-15 expression an effect that cannot be normalized by exogenous IL-1 β . (a) Flow cytometric analysis of IL-15 expression on the surface of WT and Nlrp3−/− DCs differentiated from BM progenitor cells for 10 days with Flt3L (100 ng/mL), expressed as a percentage of IL-15 expressing cells (a) and MFI per cell (b). (c) Flow cytometric analysis of IL-15 surface expression on WT and Nlrp3−/− DCs differentiated from BM progenitor cells for 10 days with Flt3L (100 ng/mL) and treated with IL-1 β (100 ng/mL) and/or recombinant IL-1 receptor antagonist (IL-1Ra; 100 μ g/mL) for 12 hours. ∗ denotes p < 0.05 compared to WT mice; # denotes p < 0.05 compared to WT, NoRx; $ denotes p < 0.05 compared to WT-IL-1 β ; n = 5/group.

Article Snippet: Sections were incubated with anti-mouse foxp3 (BioLegend Cat.# 320001; San Diego, CA, USA) or biotin-conjugated anti-mouse IL-15 (R&D Systems Cat.# BAF 447; Burlington, ON, CAN).

Techniques: Derivative Assay, Expressing, Recombinant

Journal: Mediators of Inflammation

Article Title: Exaggerated IL-15 and Altered Expression of foxp3+ Cell-Derived Cytokines Contribute to Enhanced Colitis in Nlrp3−/− Mice

doi: 10.1155/2016/5637685

Figure Lengend Snippet: A neutralizing anti-IL-15 antibody normalizes the sensitivity of Nlrp3−/− mice to DSS and reduces colonic IL-17 expression. (a) Changes in body mass in WT mice, Nlrp3−/− mice treated with 5 μ g IgG isotype (iso) control, or Nlrp3−/− mice treated with an anti-IL-15 ( α -IL-15) antibody through a 7-day course of DSS (3.5% w/v). ∗ denotes p < 0.05 compared to WT and Nlrp3−/− α -IL-15; n = 8/group. (b) Representative hematoxylin and eosin stained colonic sections (scale bar = 100 microns) and (c) blind assessment of tissue damage and inflammation in WT mice, Nlrp3−/− mice treated with 5 μ g IgG isotype (iso) control, or Nlrp3−/− mice treated with an anti-IL-15 ( α -IL-15) antibody on day 7 of DSS. ∗ denotes p < 0.05 compared to WT mice; # denotes p < 0.05 compared to Nlrp3−/− mice treated with the IgG isotype control antibody; n = 8/group. (d) Assessment of colonic tissue MPO levels and (e) colonic foxp3+ cells/HPF in WT mice, Nlrp3−/− mice treated with an IgG isotype (iso) control antibody, and Nlrp3−/− mice treated with a neutralizing anti-IL-15 ( α -IL-15) antibody on day 7 of DSS. ∗ denotes p < 0.05 compared to WT mice; # denotes p < 0.05 compared to Nlrp3−/− mice treated with the IgG isotype control antibody; ## denotes p < 0.005 compared to Nlrp3−/− mice treated with the IgG isotype control antibody; n = 8/group. (f) The expression of IL-10, IL-12p40, and IL-17 in colonic tissue isolated from Nlrp3−/− mice treated with an IgG isotype (iso) control antibody and Nlrp3−/− mice treated with a neutralizing anti-IL-15 ( α -IL-15) antibody on day 7 of DSS. ∗ denotes p < 0.05 compared to Nlrp3−/− mice treated with an IgG isotype (iso) control antibody; n = 8/group.

Article Snippet: Sections were incubated with anti-mouse foxp3 (BioLegend Cat.# 320001; San Diego, CA, USA) or biotin-conjugated anti-mouse IL-15 (R&D Systems Cat.# BAF 447; Burlington, ON, CAN).

Techniques: Expressing, Staining, Isolation