FIG. 8.

The antigen-specific response was elevated in γ/δ TCR KO mice as evidenced both by increased frequencies of cytokine-producing cells and by increased amounts of secreted cytokine after culture with Listeria antigens. PEC were harvested and pooled (five mice/group) from B6 control and γ/δ TCR KO mice 10 days following an i.p. infection with 2.4 × 10⁴ CFU of Listeria. They were incubated for 5 h with brefeldin A (BFA) in either medium alone or with 10⁷ HKLM/ml or with thioglycolate-elicited peritoneal macrophages infected with live Listeria. Cells were stained with anti-CD4- or anti-CD8-PerCP and then fixed and permeabilized and stained with anti-IFN-γ–PE and anti-IL-2–APC prior to analysis by flow cytometry. Samples were gated on CD4⁺ or CD8⁺ lymphocytes (A). Numbers in the upper right corner, frequencies of cells in each quadrant of that plot. Culture supernatants harvested after 24 h from parallel cultures without BFA were analyzed by ELISA for IFN-γ (B) and IL-2 (C). Data are representative of two independent experiments.