Th1 and Th2 cytokine production by sooty mangabey NKT clones.

<p>A) Intracellular cytokine staining of one representative NKT clone with antibodies against IFN-γ, TNF-α, IL-2, and IL-13 following 16 hours <i>in vitro</i> stimulation with PMA/Ca or the specific ligand α-GalCer presented on C1R.d cells (C1R.d/α-GC). Medium alone (R10) served as a negative control. Dot plots gated on NKT lymphocytes show intracellular expression of recent activation marker CD69 versus each cytokine with stimulation conditions denoted on the right of each panel. Numbers denote the % of NKT cells in each quadrant. B) Mean frequency of cytokine-positive NKT cells. Data on 9 NKT clones shown. Error bars denote SEM. ND: Not determined. C) Cytokine ELISA for IFN-γ, IL-4, IL-13, and IL-10 on culture supernatants collected at 24 hours post-stimulation. Mean and SEM of four clones shown. D) Cytokine profile of the NKT clones with regards to the concurrent production of IFN-γ, IL-2, and IL-13. Data on 9 NKT clones after 16 hours of stimulation shown. Pie charts (top panel) showing the proportion of one-, two-, and three-functional responses in cytokine-secreting 6B11⁺ cells. Bar charts (bottom panel) showing the mean proportion of responding cells for each of seven functional combinations. Boxes represent interquartile ranges. E) Representative dot plots (on the left) gated on NKT clones showing CD107a surface expression in CD1d/α-GalCer stimulated NKT lymphocytes in comparison to cells stimulated with C1R/α-GalCer or medium alone for 4 hours. Mean frequency of NKT clones degranulating after 4-hour stimulation (on the right). Data on surface upregulation of CD107a on four NKT cell clones shown. Error bars denote SEM.