Bronchospasm and rhinitis induced in allergic patients by in vitro released mediators. I. Methodology

This paper reports the development of an in vitro model of mediator production to be applied to patients with allergic asthma or rhinitis. All the selected patients were atopics and exhibited type I hypersensitivity to Candida alb., D. pter., or both. Heparinized blood was taken. The lymphomonocytes were separated and challenged in vitro with the specific antigen. After two hours of incubation and careful washing, the cells were further cultured for 24 hours and the supernatants removed. Skin tests and bronchial provocation tests were done with supernatants with known concentration of proteins. As a control for cell activation, two lymphokines, i.e. MIF and MF were determined in supernatants. The results demonstrated a dose-related bronchial and nasal response in allergic individuals with specific supernatants. On the other hand they failed to respond to supernatants recovered from non-sensitized cells, cells sensitized with unrelated antigens or sensitized cells obtained from non-allergic individuals. In almost 50% of the cases the immediate bronchial response, which was always present was sustained and persisted for several hours. Active supernatants always induced strong immediate skin responses. In conclusion, this in vitro model of mediator production seems to be valuable for the analysis of the physiopathological mechanism of these diseases.