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. 2023 Oct 17;26(12):108225. doi: 10.1016/j.isci.2023.108225

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© 2023 The Author(s)

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

PMC Copyright notice

CD4⁺LAG3⁺T cells improved bleomycin-induced pulmonary fibrosis in mice

(A) Experiment protocol for pulmonary fibrosis model.

(B, C, and E) Morphological staining and ELISA results show that compared with Bleomycin group, tail vein injection of CD4⁺LAG3⁺T cells or TGF-β3 improved mouse lung pathological morphology (200X) and reduced hydroxyproline content, while antagonistic TGF-β3 had the opposite effect. Data are represented as mean ± SD. Statistical significance was analyzed by one-way ANOVA (n = 5), ∗p < 0.05, ∗∗p < 0.01.

(D and F) ELISA and flow cytometry results showed respectively, compared with the control group, Bleomycin group of peripheral blood TGF - beta 3 content decreased significantly, CD4⁺LAG3⁺T cells in peripheral blood and spleen ratio decreased significantly. Data are represented as mean ± SD. Statistical significance was analyzed by unpaired two-tail t-test (n = 5), ∗p < 0.05.

(G) Immunofluorescence results showed that compared with Bleomycin group, the expression of CD4⁺LAG3⁺T cells in lung tissue were significantly increased after tail vein injection.

(H) Immunofluorescence results showed that the expression of α-SMA and CD68 in Bleomycin group was significantly higher than that in the control group, and the expression of MMT markers was downregulated after the treatment of CD4⁺LAG3⁺T cells or TGF-β3, and up-regulated after the antagonism of TGF-β3.

(I) Immunofluorescence results showed that the expression of α-SMA and fibronectin in Bleomycin group was significantly higher than that in the control group, and the expression of mesenchymal markers was downregulated after the treatment of CD4⁺LAG3⁺T cells or TGF-β3, and upregulated after the antagonism of TGF-β3.