Mtb specific HLA-E restricted T-cell clones produce Th2 cytokines.

<p>A. T-cell clones were cultured and RNA was isolated for gene-expression measurement using dcRT-MLPA, data are normalized to GAPDH as housekeeping gene (left panel); clones were stimulated for 24 hours with αCD3/28 beads before supernatants were collected to determine their maximum cytokine secretion profiles (in pg/ml) using multiplex bead arrays (middle panel); similarly, cells were stimulated for 16 hours with αCD3/28 beads in the presence of brefeldin A followed by intracellular cytokine staining to determine intracellular cytokine levels (right panel). Data are expressed as % of the CD3⁺CD8⁺ T-cell population. Data are coloured according to the amount of the molecules detected, according to the legend in the figure. Nt = not tested. B. HLA-E restricted CD8⁺ T-cell clones were stimulated for 24 hours with αCD3/28 T-cell activator beads and stained intracellular for IL-4 or IL-5 or IL-13 as well as IFN-γ. C. T-cell clones were cultured with peptide pulsed macrophages to assess their specific cytokine production in response to peptide presented by professional antigen presenting cells (top panel), or with BCG infected macrophages to assess their specific cytokine production induced by naturally presented antigen during in vitro mycobacterial infection (bottom panel). Supernatants were collected and cytokine/ chemokine levels were determined using multiplex bead arrays.