Figure 5.

Results of adoptive of CD4⁺ T cells from young (8 weeks), middle-aged (6 to 9 months), and old (24 months) C57BL/6 mice to 8-week-old RAG1KO recipients. Mice subjected to DS for 5 days were used as positive controls. A: Representative images of conjunctiva sections stained with PAS used to generate the bar graph in B. B: Number of PAS⁺ conjunctival goblet cells counted in paraffin-embedded sections expressed as number per millimeter. Bar graphs show the means ± SD of two independent experiments with two to three animals per age and sex, yielding a final sample of five right eyes per age and sex). Both eyes/animals were independently evaluated, and results were averaged. C: Representative images of conjunctiva frozen sections immunostained for CD4 (in red) used to generate the bar graph in D. D: CD4⁺ T cells infiltrating the conjunctival epithelium. Bar graphs show the means ± SD of two independent experiments with two to three animals per age and sex, yielding a final sample of five right eyes for each group). Both eyes/animals were independently evaluated, and results were averaged. E: Representative images of LG frozen sections immunostained for CD4 (in red) used to generate the bar graph in F. F: CD4⁺ T cells in the LG. Bar graphs show means ± SD of two independent experiments with two to three animals per age and sex, yielding a final sample of five individual left LGs per group and age). ^∗P < 0.05, ^∗∗P < 0.01, and ^∗∗∗P < 0.001 for within strain comparison. ^††P < 0.01 for sex comparison. Original magnification: ×40 (A, C, and E). CJ, conjunctiva; DS, desiccating stress; DS5, desiccating stress for 5 days; LG, lacrimal gland; RAG1KO, recombination activating gene-1 knockout; 8W, 8 weeks; 6-9M, 6 to 9 months; 24M, 24 months. Dashed lines indicate female-to-female comparison; solid lines indicate male-to-male comparisons.