title: Two-dimensional electrophoresis protein profiling and identification in rat bronchoalveolar lavage fluid following allergen and endotoxin challenge.
abstract: The protein content of bronchoalveolar lavage fluid (BALF) from actively sensitised Brown Norway (BN) rats challenged with allergen (ovalbumin, OA) and from naïve Brown Norway rats challenged with endotoxin (lipopolysaccharide, LPS) was analyzed and compared to healthy controls treated with vehicle only. BALF proteins were analyzed by one-dimensional (1-D) and two-dimensional (2-D) gel electrophoresis and identified by peptide mass fingerprinting matrix assisted laser desorption/ionization-mass spectrometry (MALDI-MS) or nanoliquid chromatography-tandem MS (nanoLC-MS/MS) after in-gel trypsin digestion of selected 2-D gel spots. Our study shows that the BALF protein profile is significantly different in animals after allergen (OA) or endotoxin (LPS) challenge as compared to controls, concerning the content of proteins derived from plasma or produced locally in the lung. In both challenges the following proteins presented patterns which differed qualitatively compared to control: T-kininogen I and II, alpha-1-antitrypsin, calgranulin A, fetuin A and B, and haptoglobin. Other proteins were diminished in both challenges, such as Clara cell 10 kDa secretory protein (CC10) and pulmonary surfactant associated protein B (SP-B); c-reactive protein increased in the OA-challenge and decreased in the LPS-challenge, and pulmonary surfactant associated protein A (SP-A) was decreased in the OA-challenge and was not significantly changed in the LPS-challenge. The identified proteins could be important not only for the diagnosis but have also interesting implications for medical treatment of lung inflammatory conditions. Furthermore, even if based on a limited number of animals, our results are of interest for the identification of lung protein markers and a better understanding of the mechanisms involved in the pathogenesis of lung diseases.
authors.affiliation: Novartis Institutes for Biomedical Research, Central Technologies-Analytical & Imaging Sciences Unit, CH-4002 Basel, Switzerland.
authors.forename: Luca
authors.initials: L
authors.lastname: Signor
authors.forename: Bruno
authors.initials: B
authors.lastname: Tigani
authors.forename: Nicolau
authors.initials: N
authors.lastname: Beckmann
authors.forename: Rocco
authors.initials: R
authors.lastname: Falchetto
authors.forename: Markus
authors.initials: M
authors.lastname: Stoeckli
mesh_terms: D000485:Allergens / Q000378:metabolism*; D000818:Animals; D001992:Bronchoalveolar Lavage Fluid; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D004731:Endotoxins / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000737:chemistry / Q000378:metabolism; D008168:Lung / Q000378:metabolism; D008297:Male; D051381:Rats; D011914:Rats, Inbred BN; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013997:Time Factors; D014357:Trypsin / Q000494:pharmacology
chemical_list: D000485:Allergens; D004731:Endotoxins; D008070:Lipopolysaccharides; D014357:Trypsin