FIGURE 4.
Expression of GARP/LAP on TGF-β1-, Furin-, and GARP-deficient Tregs. Staining of GARP/LAP CD4+Foxp3+ gated cells from lymph nodes or on CD4-enriched cells after activation with plate-bound anti-CD3 and IL-2 in serum-free media. Cells are from (A.) tgfb1fl/fl x CD4-CRE, (C) furinfl/fl x CD4-CRE, and (D) lrrc32fl/fl x CD4-CRE (GARP deficient). (B.) CD4+ T-cells from tgfb1fl/fl x CD4-CRE were stimulated as in (A) for 48 hours, then incubated in fresh media with (black lines) or without (grey lines) recombinant human latent TGF-β1 for 30′ at 37°C, then thoroughly washed and stained with anti-human LAP. Gating on CD4+Foxp3- cells (left panel) or CD4+Foxp3+ cells (right panel).