Quantitative proteomics using tandem mass tags in relation to the acute phase protein response in chicken challenged with Escherichia coli lipopolysaccharide endotoxin

Horvatić, A., Guillemin, N., Kaab, H., McKeegan, D. , O'Reilly, E., Bain, M. , Kuleš, J. and Eckersall, P. D. (2019) Quantitative proteomics using tandem mass tags in relation to the acute phase protein response in chicken challenged with Escherichia coli lipopolysaccharide endotoxin. Journal of Proteomics, 192, pp. 64-77. (doi: 10.1016/j.jprot.2018.08.009) (PMID:30114510)

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The inflammatory response in chickens (Gallus Gallus domesticus) is an integral part of the bird's response to infection. Detailing proteomic changes occurring during infection would be beneficial to the poultry industry, offering opportunities for comparative pathophysiological analysis. The objective of this study was to quantify the changes in the plasma proteome in chickens challenged with lipopolysaccharide (LPS), a bacterial endotoxin known to stimulate the host innate immune system. Plasma from chicken (N = 6) challenged with Escherichia coli (LPS) (2 mg/kg body weight) was collected pre (0 h) and at 12, 24, 48, and 72 h post-injection along with plasma from a control group (N = 6) challenged with sterile saline. Samples were analysed by a quantitative Tandem Mass Tags approach using a Q-Exactive-Plus mass-spectrometer. Identification and relative quantification were performed using Proteome Discoverer, and data were analysed using R. Gene Ontology terms were analysed by Cytoscape based on the Gallus gallus database. Finally, 87 significantly regulated proteins were found, including serum-amyloid-A, ovotransferrin and alpha-1-acid-glycoprotein, showing a significant effect of time post-injection in the LPS-treated group. Different pathways related with protein activation cascade and heterotopic cell-cell adhesion were affected by LPS-challenge. LPS-challenged chickens demonstrate significant changes to the plasma proteome with both increases and decreases of individual proteins within 12 h of challenge. Significance: The injection of chicken with bacterial lipopolysaccharide followed by sequential plasma and clinical analysis of the bird, is a long established and a widely used model for inflammation and infection studies. This study, utilising and combining proteomic and immunoassay analysis with bioinformatic analysis, revealed that several biological pathways are modulated during this early period of inflammation. In addition, proteins with biomarker potential were identified and successfully validated. This experimental model also demonstrated potential for pathophysiological mechanism investigation and as an inflammatory model for biomedical research. There is, despite plasma being an easily accessible biological matrix which is representative of the health status of the bird, scarce data on the chicken plasma proteome. This research makes a positive contribution to the current field, generating significant data for continuing comparative analysis.

Item Type:Articles
Additional Information:This work was supported by the European Commission FP7 “VetMedZg” Project (grant number 621394) (AH, NG, JK and PDE). The authors gratefully acknowledge Kufa University and Iraqi Ministry of Higher Education for a PhD scholarship for HK at University of Glasgow and funding from the Biotechnology and Biological Science Research Council BBSRC (BB/H016171/1) for support of E O'R.
Keywords:Broiler chicken, Innate immunity, lipopolysaccharide endotoxin, pathway analysis, TMT-based proteomics.
Glasgow Author(s) Enlighten ID:Eckersall, Professor David and O'Reilly, Dr Emily and McKeegan, Dr Dorothy and Bain, Professor Maureen and KAAB, Haider and Kules, Mrs Josipa
Authors: Horvatić, A., Guillemin, N., Kaab, H., McKeegan, D., O'Reilly, E., Bain, M., Kuleš, J., and Eckersall, P. D.
College/School:College of Medical Veterinary and Life Sciences > School of Biodiversity, One Health & Veterinary Medicine
Journal Name:Journal of Proteomics
ISSN (Online):1876-7737
Published Online:13 August 2018

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
542011Biomarkers of the innate immune response to disease in chickens: acute phase proteins and resistance to diseasePeter EckersallBiotechnology and Biological Sciences Research Council (BBSRC)BB/H016171/1III - PARASITOLOGY