Figure 1. CS particles stimulated CD4+TRM cell emergence and expansion along with silicosis progression.
(A) Schematic showed CD45-APC-Cy7 antibody intravenous (i.v. ) injection to distinguish tissue-resident or circulating leukocytes. Circulating leukocytes were labeled. (B) Flow cytometry (FC) analysis of pulmonary CD4+ TRM cells (CD45injectCD44+) and TEM (CD45inject+CD44+) cells. Percentages and counts of the CD4+ TRMcells were compared at the indicated time points (n = 5). W, week. (C) Representative FC analysis of CD4+ TRM cells for CD69, CD103 and CXCR6 expression respectively. (D) FC analysis of CD4+TRM cells for CD69 and CD103 expression. The graph showed percentages and counts of CD69+CD103 (upper) and CD69+CD103+ subsets (lower) in CD4+ TRM cells of saline or CS-treated mice at the specified time points (n = 5). (E) FC analysis of lung circulating TEM cells for CD69 and CD103 expression of saline or CS-treated mice at specified time points. The bar graph illustrated ratios of CD69+ in circulating CD4+ TEMcells (n = 4). (F) FC analysis of splenic TEM cells for CD69 and CD103. The graph compared percentages of CD69+ on splenic CD4+TEM cells of saline or CS-treated mice (n = 6). The bar graphs are the combined results of at least three independent experiments. Individual mice are plotted on the graphs. Values are reported as the mean ± SD. P value was determined by one-way ANOVA followed by Tukey’s test.