Figure 6. Suppressive activity of TNF-α-producing CD4+CD25+Foxp3+ Treg cells.
(A and B) Expression levels of proteins known to be related with the Treg cell suppression function, including Foxp3 (A) and CD39 (B), were analyzed by flow cytometry. (C) Anti-CD3/CD28-stimulated proliferation of non-Treg CD4+ (left) and CD8+ (right) responder T cells was assessed by measuring the percentage of CFSElo cells in Treg-depleted PBMCs and Treg-reconstituted PBMCs at various ratios. Reconstitution at the original ratio of Treg and non-Treg cells in the peripheral blood is marked as ‘1:1’ and indicated by the arrow. The percentage of suppression was calculated as [1-(% Tresp proliferation in the reconstituted PBMCs/% Tresp proliferation in the Treg-depleted PBMCs)]×100 and was compared between AHA patients (n=11) and healthy controls (n=8). (D) Anti-CD3/CD28-stimulated proliferation of non-Treg CD4+ (left) and CD8+ (right) responder T cells was assessed by measuring the percentage of CFSElo cells in Treg-depleted PBMCs and Treg-reconstituted PBMCs from AHA patients (n=5) and healthy controls (n=5) in the presence or absence of anti-TNF-α blocking antibody. The percentage of suppression was calculated as shown above. Error bars represent standard deviations.
