Figure 2

Stimulation of cells by L. acidophilus strains. Excised colons from untreated (PBS) mice (n = 5/group) and mice (n = 5/group) treated with NCK2031, NCK56, NCK2025, or S. aureus LTA were stained with CD11c, TNFα, IL-12, or CD4, IFNγ, and visualized by UV laser scanning microscope. White size bar represents 20 μm. White arrows point to the co-localized cells in yellow dots. (a) Representation of CD11c⁺/TNFα⁺, CD11c⁺/IL-12⁺ cells (yellow, merged red/green/blue). White size bar represents 50 μm. White arrows point to the co-localized cells in yellow dots. (b) Quantitative analyses of IL-12 and TNFα⁺ by TissueGnostics Tissue/Cell High-Throughput Imaging and Analysis System. (c) Induction of colonic DC-activation by L. acidophilus expressing LTA. Single cells were derived from colons of the mice (n = 5/group) treated with NCK20331, NCK56, NCK2025, or PBS. Cells were stained with anti-CD11c, CD11b, CD103, MHC II, CD19, F4/80, IL-12, or TNFα antibodies. Live CD11c⁺ cells that were F4/80^-CD19^- were analyzed for their cytokine expression and expression of surface molecules (i.e., MHC II) by FACSCanto II. Experiments were repeated at least three times with similar outcomes.