Identification of immuno-reactive proteins from a sheep gastrointestinal nematode, Trichostrongylus colubriformis, using two-dimensional electrophoresis and mass spectrometry

• Published in: International journal for parasitology 2007-11, Vol.37 (13), p.1419-1429
• Main Authors: Kiel, Markus, Josh, Peter, Jones, Alun, Windon, Ross, Hunt, Peter, Kongsuwan, Kritaya
• Format: Article
• Language: English
• Subjects: 2DE; animal proteins; Animals; antigen-antibody reactions; antigens; Antigens, Helminth - immunology; Antigens, Helminth - isolation & purification; Biological and medical sciences; Blotting, Western; electrophoresis; Fundamental and applied biological sciences. Psychology; Gastrointestinal Diseases - immunology; Gastrointestinal Diseases - parasitology; Gastrointestinal Diseases - veterinary; gastrointestinal nematodes; IgG; immune response; Immuno-reactive; Immunoelectrophoresis, Two-Dimensional; Immunoglobulin G; Invertebrates; Life cycle. Host-agent relationship. Pathogenesis; Mass Spectrometry; Nemathelminthia. Plathelmintha; Nematoda; nematode infections; Nematode Infections - immunology; Nematode Infections - parasitology; Nematode Infections - veterinary; Nematodes; parasite antigens; protein identifciation; Proteomics; Protozoa; Sheep; Sheep Diseases - immunology; Sheep Diseases - parasitology; soluble antigens; Trichostrongylus; Trichostrongylus - immunology; Trichostrongylus - isolation & purification; Trichostrongylus colubriformis; two-dimensional gel electrophoresis
• ISSN: 0020-7519; 1879-0135
• DOI: 10.1016/j.ijpara.2007.04.016

Gastrointestinal nematode infections of livestock animals are prevalent and costly problems worldwide. Currently, infections are controlled by anthelmintic chemicals but increasing drug resistance has prompted research interest to shift towards alternative methods of control such as vaccine development and selection of worm-resistant animals. The present study analyses proteins from Trichostrongylus colubriformis infective L3s that are recognised by IgG of immune sheep. Following protein separation via two-dimensional electrophoresis and Western blot probing with plasma from sheep resistant to T. colubriformis, mass spectrometry-based proteomic analyses were used to identify immuno-reactive protein spots. We were able to identify 28 immune targets, including aspartyl protease inhibitor, enolase, chaperone proteins, galectin, glycolytic enzymes, kinase, phosphatase and structural muscle proteins such as myosin, paramyosin, calponin and DIM-1. The data suggest that immune responses to T. colubriformis are dispersed over a relatively large number of parasite antigens, including several cytoplasmically expressed proteins. The results have new implications for understanding the molecular mechanisms that underpin host–parasite interaction during gastrointestinal nematode infections.