Pulmonary Humoral Immune Response
Pulmonary immune response was evaluated by measurements of the levels of IL-2, IL-10, IL-17 and IFN-gamma in the breath condensate by DuoSet ELISA kits (R&D Systems; MN, USA). Breath condensate was collected for 10 minutes using RT Tube (Respiratory Research, TX, USA) according to the manufacturer’s recommendations and immediately stored in liquid nitrogen and further transferred to -86°C freezer, until the measurements were done, using a microplate reader Spectramax I3 (Molecular Devices, CA, USA).