Repeated Inhalation Challenge with Diphenylmethane-4,4′-Diisocyanate in Brown Norway Rats Leads to a Time-Related Increase of Neutrophils in Bronchoalveolar Lavage After Topical Induction

Abstract

Diphenylmethane-4,4′-diisocyanate (MDI) is a low-molecular-weight chemical known to cause occupational asthma. The objective of this study was to evaluate the topical and inhalation routes of sensitization on the elicitation response of MDI in the Brown Norway (BN) rat model following repeated challenge exposures. BN rats were either induced topically (150 μl MDI on the flanks, booster administration to the skin of the dorsum of both ears using 75 μl/dorsum of each ear) or by inhalation (5 × 3 h/d, 28.3 ± 3.0 mg MDI/m³ [± SD]). Inhalation challenge exposures with MDI (15.7 ± 1.4 mg/m³, duration 30 min) were made on d 21, 35, 50, and 64. One day after each challenge, rats were rechallenged with methacholine (MCh) aerosol. Respiratory changes were monitored during challenges. One day after the MCh challenge, selected endpoints in bronchoalveolar lavage (BAL), the weights of lungs, and auricular and lung-associated lymph nodes were determined. After the first and last challenge, lymph nodes and lungs were examined by histopathology. Repeated challenge with MDI or MCh did not elicit marked changes in respiratory patterns at any time point. Mild but consistent time-related increased BAL neutrophils and slightly increased lung and lymph-node weights occurred in topically sensitized rats as compared to the remaining groups. In topically sensitized rats, in the lung histopathology revealed activated lymphatic tissue and an increased recruitment of airway eosinophils. Immunoglobulin (Ig) E determinations (serum and BAL) did not show any differences amongst the groups. Thus, high-dose topical induction with MDI was associated with a neutrophilic and eosinophilic inflammatory response in the lung after repeated inhalation challenge with MDI, with magnitude of effect dependent on the specific methodology used.