Figure 1. Production of inflammatory cytokines by Treg cells from AHA patients.

(A–C) PBMCs from AHA patients were stimulated with anti-CD3/CD28 antibodies, and cytokine production (TNF-α, IFN-γ, and IL-17A) was analyzed by ICS. Percentages of cells producing each cytokine among CD4⁺CD25⁺CD127^lo/−Foxp3⁺ T cells from AHA patients were determined (n=19 for TNF-α and IFN-γ; n=11 for IL-17A) and compared to those from healthy controls (n= 19 for TNF-α and IFN-γ; n=14 for IL-17A) (A). Horizontal bars represent mean percentages. Representative flow cytometry dot plots of a patient and a healthy donor are shown in (B). Plots are gated on CD4⁺CD25⁺CD127^lo/−Foxp3⁺ T cells. Mean fluorescence intensity of TNF-α in TNF-α-producing CD4⁺CD25⁺Foxp3⁺ T cells is compared between AHA patients (n=19) and healthy controls (n=19) (C). (D) PBMCs from AHA patients (n=4) were stimulated with HAV VP2 protein, HAV VP2 OLP mix or anti-CD3/CD28 antibodies, and TNF-α production was analyzed by ICS. Percentage of TNF-α-producing cells among CD4⁺CD25⁺CD127^lo/−Foxp3⁺ Treg cells was presented. (E) PBMCs from AHA patients (n=4) were stimulated with HAV VP2 protein or HAV VP2 OLP mix, and TNF-α production was analyzed by ICS. In the gate of TNF-α⁺ T cells, percentages of CD4⁺CD25⁺CD127^lo/−Foxp3⁺ Treg cells, Foxp3⁻ non-Treg CD4⁺ T cells and CD8⁺ T cells were determined and presented.