Mononuclear lymphocytes from the lungs were isolated on day 14, following nasal-pulmonary immunizations of wild-type, IFN-γ−/− and IL-4−/− mice. Pooled lung lymphoid cells from 4 mice/group were cultured, in vitro, in the absence (grey bars) and presence (black bars) of mycoplasma antigen. Supernatants were collected 4 days later, and the levels of IFN-γ, IL-4, IL-5 and IL-17 were determined using a suspension array. The lowest detection level for the cytokines was 3.2 pg/ml. Detection was done in samples collected from 2 separate experiments.