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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 regulate macrophage polarization and mesenchymal transition

(A) Flow cytometry results show that adding CD4⁺LAG3⁺T cells supernatant to macrophages inhibited macrophages polarization to M1 and promoted macrophages polarization to M2.

(B) Transwell results show thatCD4⁺LAG3⁺T cells inhibited TGF-β1-induced M1 macrophage migration, while after antagonism TGF-β3 the effects were weakened. Data are represented as mean ± SD. Statistical significance was analyzed by one-way ANOVA (n = 3), ∗p < 0.05, ∗∗p < 0.01.

(C) ELISA results show that CD4⁺LAG3⁺T cells promote TGF-β1-induced M2 macrophage secretion of IL-10 and Arg1, while after antagonism TGF-β3 the effects was weakened. Data are represented as mean ± SD. Statistical significance was analyzed by one-way ANOVA (n = 3), ∗p < 0.05, ∗∗∗p < 0.001.

(D and E) PCR results show that CD4⁺LAG3⁺T cells inhibited TGF-β1 induced collagen Ⅰ and Ⅲ secretion, and inhibited the expression of interstitial markers fibronectin, vimentin, and α-SMA mRNA expression, while the effect was weakened after antagonist TGF-β3. Data are represented as mean ± SD. Statistical significance was analyzed by one-way ANOVA (n = 3), ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

(F) Western blot results show that CD4⁺LAG3⁺T cells inhibited TGF-β1 induced mesenchymal transition, while the effect was weakened after antagonist TGF-β3. Data are represented as mean ± SD. Statistical significance was analyzed by one-way ANOVA (n = 3), ∗p < 0.05, ∗∗p < 0.01.

(G) Western blot results show that CD4⁺LAG3⁺T cells regulated macrophages through non-contact secretion of TGF-β3. Data are represented as mean ± SD. Statistical significance was analyzed by one-way ANOVA (n = 3), ns = not significant, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.