One subset of DR⁻ Treg clones can suppress or secrete IL-17 in response to different stimuli. The clones that grew from wells seeded at 1 DR⁻ Treg per well were analyzed for several features and compared with clones derived from Tresp cells. After 5 weeks of expansion, a portion of each clone was stained for FoxP3 and IL-17 expression and tested for ability to suppress the proliferation of freshly isolated Tresp cells in cocultures stimulated with αCD3/αCD2 beads. On day 4, half the media in each well replaced with ³H thymidine to monitor proliferation. Data are representative of 2 independent cloning experiments. Intracellular expression of FoxP3/IL-17 (A), IL-2 (B), and IFNγ (C) of DR⁻ Treg clones after 4 hours of stimulation with PMA/ionomyicin and GolgiStop. One representative clone from each pattern is shown. (D) Mean FoxP3 expression by each clone. (E) The suppressive capacity of each clone represented as percentage suppression. (F) Mean FoxP3 expression by each clone is shown relative to its suppressive ability. (G) Suppression by each pattern of clone.