Figure 2.

Aspergillus antigens stimulate Th cells in vitro and indicate increased local Th17 cells in SEA. Cryopreserved BALC and PBMC from HE (BALC, n=9; PBMC, n=7), MEA (n=3), or SEA (BALC n=5, PBMC n=8) were restimulated in vitro for 24 h with recombinant A. fumigatus antigens (Asp f 1, or mixed Asp f 7 and Asp f 8) and evaluated for cytokine expression by flow cytometry in comparison with medium-incubated controls (net percentage). (A) An example of SEA BALC gated on singlet live lymphocytes is shown (pseudocolour with enlarged dots for visibility). (B–D) Each cytokine was evaluated in CD4⁺ Th cells within BALC and PBMC. (B) Asp f 1 stimulated higher percentages of CD4⁺IL-17⁺ Th17 cells in SEA compared with HE BALC. Asp f 7/8 stimulation yielded a similar trend. (C) Asp f 7/8 stimulated more CD4⁺IL-4⁺ Th2 cells in SEA compared with HE BALC but (D) CD4⁺IFN-γ⁺ Th1 stimulations were comparable between the groups. The stimulation of PBMC did not yield group differences in Th responses to the antigens.