Th17 cell clone generation and characterization. (A, B) Boolean gating analysis showing all combinations of IL-17A, IL-22 and IFN-γ production by CD4 + CD45RA- memory T cells before and after stepwise enrichment based on CCR6 + CCR4 + CCR10- and CD161+ surface expression (A), and in the 20 T cell clones expanded after enrichment (B). Intracellular cytokine staining was performed after PMA and ionomycin stimulation. Color codes are shown in the right panel of A. The green circular segment indicates the total number of IL-17A-producing T cells after each step of enrichment. (C) T cell cytokine levels were measured in supernatants of five of the twenty activated Th17 cell clones by ELISA and bead immunoassay.
CCR: CC-chemokine receptor; CD: cluster of differentiation; ELISA: enzyme immunosorbent assay; IFN: interferon; IL: interleukin; PMA: phorbol myristate acetate.