Figure 3.

Quantitative analyses of immune cells in mesenteric lymph nodes (MLNs) showed a reduction in CD4⁺ T-lymphocytes in NPP7 KO mice. MLNs from NPP7 HT and KO mice, and WT mice, were stained for various immune cell populations by immunohistochemistry. Computerized image analysis was used to quantify positivity (stained fraction of tissue area) reflecting cell numbers. (A) Representative images of MLNs from WT mouse (left) and NPP7 KO mouse (right), out of 32 and 46 images, respectively, illustrating the staining of CD3ε⁺ T-lymphocytes in brown (DAB chromogen); (B) CD3ε⁺, CD4⁺ and CD8α⁺ T-lymphocytes; (C) Foxp3⁺ T_regs; (D) B-lymphocytes; (E) CD138⁺ plasma cells (F) IgA⁺ cells; (G) Representative images of MLNs from WT mouse (left) and NPP7 KO mouse (right), out of 32 and 46 images, respectively, illustrating the staining of macrophages in brown (DAB chromogen) and DCs in green (Vina green chromogen) [upper panels], and quantitative image analysis of CD11c⁺F4/80⁻CD163⁻ DCs shown in green with the macrophage signal quenched [lower panels]; (H) CD11c⁺F4/80⁻CD163⁻ DCs; and (I) Macrophages. Each dot represents an individual mouse and is the mean of two staining experiments. Columns show the group mean ± SD, ***P<0.001.