Author:

Keywords:

Science & Technology, Life Sciences & Biomedicine, Biochemical Research Methods, Biochemistry & Molecular Biology, 2D-DIGE, auricular lymph nodes, BALB/c mouse, bronchoalveolar lavage, serum, toluene-2,4-diisocyanate, BRONCHOALVEOLAR LAVAGE FLUID, INDUCED OCCUPATIONAL ASTHMA, IMMUNOLOGICAL DETERMINANTS, DERMAL SENSITIZATION, TOLUENE DIISOCYANATE, T-LYMPHOCYTES, LUNG-DISEASE, GC-GLOBULIN, IN-VITRO, COMPLEMENT, OCCUPATIONAL ASTHMA, EXPRESSION, SERUM, LUNG, EXPOSURE, Animals, Asthma, Bronchoalveolar Lavage Fluid, Disease Models, Animal, Inflammation, Mass Spectrometry, Methacholine Chloride, Mice, Neutrophils, Oxidative Stress, Proteome, Toluene 2,4-Diisocyanate, 03 Chemical Sciences, 06 Biological Sciences, 31 Biological sciences, 34 Chemical sciences

Abstract:

Occupational asthma is the principal cause of work-related respiratory disease in the industrial world. Toluene-2,4-diisocyanate (TDI) is one of the most common respiratory sensitizers leading to occupational asthma. Using a mouse model of chemical-induced asthma, we explored proteome changes in multiple compartments of mice sensitized and challenged with TDI or acetone-olive oil (AOO; vehicle). Airway reactivity to methacholine and a bronchoalveolar lavage (BAL) cell count was assessed in treated and control mice, 1 day after challenge. Subsequently, two-dimensional differential gel electrophoresis (2D-DIGE) was performed on auricular lymph nodes, BAL, and serum comparing TDI-treated and vehicle-treated control mice. The differentially expressed proteins were identified by mass spectrometry and pathway analysis was performed. TDI-treated mice exhibit increased airway reactivity (2.6-fold increase) and a neutrophilic inflammation in the BAL fluid, compared to control mice. 2D-DIGE showed 53, 210, and 40 differentially expressed proteins in the auricular lymph nodes, BAL, and serum of TDI-treated versus vehicle-treated mice, respectively. Several of the identified proteins could be linked with inflammation, neutrophil chemotaxis, and/or oxidative stress. Physiologic and immunologic readouts of the asthmatic phenotype, such as inflammation, were confirmed in three compartments by several of the differentially expressed proteins via 2D-DIGE and computerized pathway analysis.