Evaluation of protein expression in bovine bronchoalveolar fluid following challenge with Mannheimia haemolytica |
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Authors: | Boehmer Jamie L Degrasse Jeffrey A Lancaster Vicki A McFarland Melinda A Callahan John H Ward Jeffrey L |
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Institution: | U.S. Food and Drug Administration Center for Veterinary Medicine, Laurel, MD 20708, USA. jamie.boehmer@fda.hhs.gov |
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Abstract: | Proteomics analysis of bovine bronchoalveolar fluid (BAF) following induction of pneumonia with Mannheimia haemolytica using nanoflow liquid chromatography coupled with tandem mass spectrometry (nanoLC-MS/MS) resulted in the identification of 88 unique proteins. Proteins detected in BAF included antimicrobial peptides (AMPs), complement factors, acute-phase proteins, protease inhibitors, and proteins involved in oxidation-reduction. Notwithstanding biological variation, differences in relative protein abundance, determined using normalized peptide counts, were detected for select proteins in BAF from genuinely infected versus sham-infected animals. To demonstrate the applicability of using normalized peptide counts to assess protein expression trends, LC-MS/MS data for the acute-phase protein haptoglobin (HPT) were compared with ELISA data, and statistical evaluation of the relationship between the data revealed a strong measure of association. Differences were detected between sham- and genuinely infected animals for haptoglobin, as well as the AMPs cathelicidin-1 and cathelicidin-4, and inter-α-trypsin inhibitor heavy chain-4, a fairly novel protein involved in the acute phase response. Though the small sample size limited the scope of the inferences, the results indicate the likely importance of AMPs and acute-phase proteins during respiratory infection, and provide additional information regarding potential mechanisms involved in the bovine mucosal barrier defense. |
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Keywords: | Animal proteomics Antimicrobial protein Bovine bronchoalveolar fluid MS/MS Nanoflow LC |
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