Figure 2.

Functional analysis of cellular adaptive immune compartment. (A) Lymphoproliferative responses of peripheral blood mononuclear cells (PBMC) from the patient (P) and control (C) individuals to different mitogenic stimuli [concanavalin A (Con A), phytohemagglutinin (PHA), Pokeweed (PKW), candidin, and antigens of Mycobacterium tuberculosis or Trychophyton spp.] or medium alone (mock). Cells were cultured (3.0x10⁵/well) in quadruplicate for each condition and incubated for 96 h at 37°C/5% CO₂ as previously described (33). During the last 18 h of culture, wells were pulsed with 1 µCi of [methyl-3H] thymidine in fresh medium. Cells were harvested onto glass fiber filters and labeled material was counted in a β scintillation counter. Results were expressed as counts per minute (cpm). (B) IFNγ, IL-17A, TNF-α and IL-10 concentrations in PBMCs culture supernatants determined by sandwich ELISA and results expressed as pg/ml. Experiments were performed at least in quadruplicate and repeated twice with similar results. Data are shown as mean ± SEM, the patient and sex and age-matched control individuals (n=6 healthy females, aged 3-7 years old). Red bars correspond to the average values of the patient and white bars correspond to the average values of the controls analyzed. Mann-Whitney test; *p< 0.05, **p< 0.01 and ***p< 0.005.