Study on the assessment of industrial respiratory allergens using the analysis of cytokine

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To develop the method for assessment of respiratory sensitivity of industrial chemicals in mice, we have examined the proper methods to induce sensitization with chemicals and the indices of respiratory allergy. Firstly, mice were sensitized with inhaled toluene diisocyanate (TDI) by intranasal route for several schedules of exposure and challenged with inhaled TDI.In addition to this method, mice were sensitized by epicutaneous route and challenged by inhalation. The analysis of cytokine secretion patterns in bronchoalveolar lavage fluid (BALF) and the supernatants in which splenocytes were cultured with PHA, and the measurement of serum total IgE antibody were carried out using sandwich ELISA.Consequently, the significant elevation of total IgE antibody was shown in mice which were sensitized with inhaled TDI in the twice of 4 consecutive days-exposure which has 6 days of rest and 7 days after last sensitization exposure. At the same time, the significant elevation of IL-4 and GM-CSF production levels in PHA-stimulated splenocytes were shown in the sensitized groups. While IFN-gamma production level elevated with the control groups 24 and 48 h after challenge. The similar patterns of cytokine secretion were also shown in mice sensitized by the epicutaneous route. Assessment of cytokine secretion patterns in BALF was disturbed by the significant elevation of cytokine levels in the control groups. These results indicated that the sensitization by intranasal or epicutaneous route and the challenge by intranasal route about 20 days after the first sensitization exposure was useful. The analysis of cytokine, IL-4, GM-CSF, and IFN-gamma, secretion patterns in PHA-stimulated splenocytes using sandwich ELISA was also suggested to be available for the indices of assessment of chemicals causing respiratory allergy.