Fig. 2: Pulmonary CD4+ TRM cells mediated severe lung inflammatory response to CS particles. | Communications Biology

Fig. 2: Pulmonary CD4+ TRM cells mediated severe lung inflammatory response to CS particles.

From: Crystalline silica-induced recruitment and immuno-imbalance of CD4+ tissue resident memory T cells promote silicosis progression

Fig. 2

a The reconstitution sketch of Rag1–/– mice with different cell origins. The MACS-purified CD4+ T cells and PBS were i.v. transferred into Rag1–/– mice before CS instillation. The recipient mice were analyzed one week after the CS instillation. b H&E staining to the lung sections of distinct reconstituted Rag1–/– mice. Scale Bar = 500 μm (upper) and 200 μm (lower). c The inflammation scores were assessed in the lung sections. d Relative RNA levels of Ifng, Tnfa, Il17a, and Il6 in each group. e FC analysis of pulmonary resident CD4+ T cells in transferred Rag1–/– mice. f Flow histogram indicated Ki-67 intensity in lung-resident CD4+ T cells. g FC analysis compared the proportions of CD103– to CD103+ in the CD69+ CD4+ TRM cells. h The scheme indicated the time points of CS instillation: Single CS instillation was treated at 0 W (3 mg in 50 μL), while 50 μL sterile saline per week was instilled at 1 and 2 weeks. Repeated CS instillation was treated at 0, 1, and 2 W (1 mg in 50 μL per week), respectively. Both groups were sacrificed 10 W after the first CS treatment, with an equal amount of CS particles in total. i Masson’s trichrome staining to the lung sections of different groups. Scale bar = 500 and 200 μm. The bar graph showed the fibrosis-positive area. j Relative mRNA levels of Col1a1 and Fn in each group. k FC analysis of pulmonary CD4+ TRM cells in single or repeated CS-treated mice. l Relative mRNA levels of Il17a, Ifng, and Tnfa in each group. m FC analysis showed the ratios of CD103– to CD103+ subsets in CD69+ CD4+ TRM cells. For panels a–g, n = 4–5 biologically independent animals, n number also indicated independent experimental replicates. Littermate recipient Rag1–/– mice and donor C57BL/6J mice were used in these experiments. Values are reported as mean ± SD. P value was determined by one-way ANOVA followed by Tukey’s test. For panels, h–m, n = 6–7 biologically independent animals, the bar graphs are the combined results of at least three independent experiments. Individual mice are plotted on the graphs. Values are reported as mean ± SD. P value was determined by unpaired two-tailed Student’s t-test.

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