Figure 2.

Promotion of STZ-induced immune cell infiltration into islets by the deficiency of EBI3 and WSX-1. A: Representative photomicrographs of H&E- stained islets from WT, EBI3^−/−, and WSX-1^−/− mice on day 7 after treatment with control vehicle and STZ (top panel). Arrows indicate polymorphonuclear cells. Scale bar = 20 μm. Representative photomicrographs of phycoerythrin-conjugated anti-neutrophil antibody 7/4–stained islets from WT, EBI3^−/−, and WSX-1^−/− mice on day 7 after STZ treatment (middle panel). Arrows indicate 7/4⁺ neutrophils. Scale bar = 20 μm. The number of polymorphonuclear cells (left bottom panel) and 7/4⁺ neutrophils (right bottom panel) in islets of mice treated with STZ is shown. No significant difference in the number of polymorphonuclear cells and 7/4⁺ neutrophils in islets of mice treated with STZ was detected between three groups. B: Representative photomicrographs of islets triple stained with Alexa Fluor 488–conjugated anti-F4/80, Alexa Fluor 647–conjugated anti-CD11c and phycoerythrin-conjugated 7/4 antibodies from WT, EBI3^−/−, and WSX-1^−/− mice on day 28 after treatment with control vehicle and STZ (top panel). Scale bar = 20 μm. Insets: Representative F4/80⁺CD11c⁻7/4⁻ cells are shown. Scale bar = 5 μm. The number of F4/80⁺CD11c⁻7/4⁻ cells in islets of mice treated with STZ is shown (left bottom panel). *P < 0.05 versus WT mice. mRNA expression levels of F4/80 antigen in pancreatic tissues from STZ-treated mice that were examined by real-time PCR, normalized with respect to β-actin expression, and expressed relative to that of WT mice, are shown (right bottom panel). *P < 0.05 versus WT mice. C: Representative photomicrographs of islets double stained with FITC-conjugated anti-CD4 antibody and phycoerythrin-conjugated anti-CD8 antibody from WT, EBI3^−/−, and WSX-1^−/− mice on day 28 after treatment with control vehicle and STZ (top panel). Scale bar = 20 μm. Insets: Representative CD4⁺ T cells (green) and CD8⁺ T cells (red) are shown. Scale bar = 5 μm. The number of CD4⁺ T cells (left middle panel) and CD8⁺ T cells (left bottom panel) in islets of mice treated with STZ is shown. *P < 0.05 versus WT mice. mRNA expression levels of CD4 and CD8 in pancreatic tissues from STZ-treated mice that were examined by real-time PCR, normalized with respect to β-actin expression, and expressed relative to that of WT mice are shown (middle right and bottom right panels, respectively). *P < 0.05 versus WT mice. For quantification, 10 nonconsecutive standard islets were randomly selected from 10 sections per mouse, and the numbers of cells were counted in samples from seven mice in each group. Data shown are the mean ± SE of cell number.