B-cell infiltration in lacrimal gland (LG). (A) Representative images of CD19+ cells in the LGs of IFN-γKO, γDKO, and CD25KO mice at ages 8, 12, and 16 weeks (8W,12W,16W, respectively). CD25KO 1 and 2 are two different animals of the same age. Positive cells are stained red. Scale bar, 100 μm. (B) High magnification of black square in LG at age 16 weeks in all three strains. Scale bar, 50 μm. (C) Representative flow-cytometry plots of freshly isolated cells from LGs of different strains stained with B220-APC conjugated antibody (x axis) versus forward-scatter area (FSC-A, y axis). Lymphocytes were gated based on characteristic light-scatter properties; single lymphocytes were gated based on forward-scatter height versus forward-scatter area (FSC-A), and dead cells were excluded with propidium iodide. (D) Bar graph showing mean ± standard deviation of percentage of B220+ cells gated in LG isolates with flow cytometry at ages 8, 12, and 16 weeks (8W,12W,16W, respectively). *P < 0.05 intrastrain comparison; **P < 0.01 intrastrain comparison versus age 8 weeks.