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. 2022 Sep 20;13:892254. doi: 10.3389/fimmu.2022.892254

Figure 3.

Figure 3

Migrated LCs more efficiently prime functional Treg responses. (A) Flow cytometry analysis of Tregs induced after co-culture of steady-state and migrated LC with CD4+ naive T cells as in Fig. 2C. n=8 control, n=5 steady-state LCs and n=6 migrated LCs from independent donors. *p<0.05, **p<0.01, ***p<0.001. (B) Proliferation analysis of CD4+ T cells using CFSE labelled PBMCs after 3-day co-culture with autologous purified CD3+CD4+CD127-CD25+ Tregs. The percentages of proliferating CD4+ cells stimulated with plate bound anti-CD3 and soluble anti-CD28 are displayed at ratios of 1:1 and 1:3 Treg : PBMC (n=5 from 3 independent LC donors). *p<0.05, **p<0.01. (C) Proliferation analysis of CD8+ T cells using CFSE labelled PBMCs after 3-day co-culture with autologous purified CD3+CD4+CD127-CD25+ Tregs. The percentages of of proliferating CD8+ cells stimulated with plate bound anti-CD3 and soluble anti-CD28 are displayed at ratios of 1:1 and 1:3 Treg : PBMC (n=5 from 3 independent LC donors). *p<0.05, ***p<0.001. (D) Flow cytometry assessment of the percentage of Tregs induced after 5-day co-culture of migrated LC with autologous TRMs extracted from human epidermis. 5-day cultures of TRMs alone were used as control. Tregs were identified as CD3+CD4+CD127-CD25+FOXP3+ cells. n=5 independent LC donors. **p<0.01. (E) Percentage of IL-10 producing CD4+ cells after co-culture of TRMs in the presence or absence of migrated LC. n=8. *p<0.05, ****p<0.0001.