How to cite this article: Vet. Res. (2008) 39:58
DOI: 10.1051/vetres:2008035

Proteomic approach to identify candidate effector molecules during the in vitro immune exclusion of infective Teladorsagia circumcincta in the abomasum of sheep

Spiridoula Athanasiadou^{1, 2}, Alan Pemberton³, Frank Jackson², Neil Inglis⁴, Hugh R.P. Miller³, Frank Thévenod⁵, Annie Mackellar² and John F. Huntley<sup>2

1</sup>  Animal Nutrition and Health, Scottish Agricultural College, Bush Estate, Penicuik, EH26 0PH, Scotland, UK
²  Division of Parasitology, Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik, EH26 0PZ, Scotland, UK
³  Division of Veterinary Clinical Studies, University of Edinburgh, Easter Bush Veterinary Centre, Roslin, EH25 9RG, Scotland, UK
⁴  Moredun Proteomics Facility, Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik, EH26 0PZ, Scotland, UK
⁵  Department of Physiology and Pathophysiology, University of Witten/Herdecke, Stockumer Str. 12, D-58453 Witten, Germany

Received 26 March 2008; accepted 13 August 2008; published online 22 August 2008

Abstract - In the present study we have employed an in vitro organ challenge model to study the post-challenge responses in parasite naïve and immune gastric tissue of sheep, in an attempt to identify the host derived factors involved in immune exclusion of Teladorsagia circumcincta larvae. Proteins present in the epithelial cells and mucus from ovine abomasa following parasite challenge in previously naïve and immune animals were analysed through Matrix Assisted Laser Desorption/Ionization-Time of Flight (MALDI-Tof)-MS and shotgun proteomics. MALDI-ToF analysis of epithelial cell lysates revealed that a number of proteins identified were differentially expressed in naïve and immune cells. These included intelectin and lysozymes, which were present at higher levels in epithelial cell lysates derived from immune samples. A large number of proteins were identified in the mucosal wash from immune tissue which were not present in the mucosal wash of the naïve tissue. Some of these proteins were present in washes of immune tissue prior to the parasite challenge including immunoglobulin A, galectin 14 and 15 and sheep mast cell protease 1. However, other proteins, such as calcium activated chloride channel and intelectin were only detected in the washings from the challenged tissue. The latter may be related to an enhanced mucus release, which may result in entrapment of infective larvae and thus reduced establishment in tissue that has been previously challenged with the parasite. In conclusion, several proteins have been identified which may be involved, either directly or indirectly, in the exclusion and immune elimination of incoming infective larvae. In the present study, the usefulness of the in vitro model has been confirmed, and the global proteomic approach has identified proteins that had not previously been associated with parasite exclusion from abomasal mucosa, such as the calcium activated chloride channel.

Key words: immunity / mucus / proteomics / sheep / Teladorsagia circumcincta

Corresponding author: spiridoula.athanasiadou@sac.ac.uk

© INRA, EDP Sciences 2008