FIGURE 7.

EmACT promotes host TGF-beta-dependent Foxp3⁺ Treg conversion in vitro. Freshly generated BMDCs (Day 8) were co-cultured with naïve (CD25^–) OT-II.RAG-1^–/^– CD4⁺ T-cells at a DC:T-cell ratio of 1:3 in R10 medium supplemented with OVA peptide (200 ng/ml) in the presence of supernatant from pSecTag2-transfected HEK (Control) or pSecTag2-emact-transfected HEK (rEmACT) supplemented or not with rhTGF-β1 (1 ng/ml). After 5 days of incubation, cells were harvested and stained for CD4, CD25 and Foxp3 prior to flow cytometry analysis. (A) Representative plots of two independently performed Treg conversion assays with two different DC/T cell preparations with supernatant from 2 batches of transfected HEK cells summarized in (B). The bars represent the mean ± SD.