Figure 1

Example of allergen specific Th and B cells. Peripheral blood MNCs (in this example from Timothy-allergic individual per skin prick test) labeled with carboxyfluorescein diacetate succinimidyl ester (CFSE) were cultured for 7 days with Timothy allergen. Glycerol saline (negative control) and anti-CD3 (positive control) were used as controls. Monensin was used to stop cytokine secretion. At the end of the culture, cells were stained for CD3, CD4, CD19, CD20, IFNg and IL-4 and analyzed using FACS Aria flow cytometer. Timothy-specific Th, Th1, Th2 and B cells are defined as CFSE^lowCD3^posCD4^pos (P1), CFSE^lowCD3^posCD4^posIFNg^pos (P2), CFSE^lowCD3^posCD4^pos IL-4 ^pos (P3) and CFSE^lowCD3^pos(CD19 ^pos or CD20 ^pos) (P4) cells, respectively.