Open Access Research

A novel strategy for the identification of antigens that are recognised by bovine MHC class I restricted cytotoxic T cells in a protozoan infection using reverse vaccinology

Simon P Graham1*, Yoshikazu Honda1, Roger Pellé1, Duncan M Mwangi1, E Jane Glew1, Etienne P de Villiers1, Trushar Shah1, Richard Bishop1, Pierre van der Bruggen3, Vishvanath Nene2 and Evans LN Taracha1

Author Affiliations

1 International Livestock Research Institute, P. O. Box 30709, Nairobi 00100, Kenya

2 The Institute for Genomic Research, 9712 Medical Center Drive, Rockville, MD 20850, USA

3 Ludwig Institute for Cancer Research – Brussels branch, Avenue Hippocrate 74 – UCL 7459, B-1200 Brussels, Belgium

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Immunome Research 2007, 3:2  doi:10.1186/1745-7580-3-2

Published: 9 February 2007

Abstract

Background

Immunity against the bovine protozoan parasite Theileria parva has previously been shown to be mediated through lysis of parasite-infected cells by MHC class I restricted CD8+ cytotoxic T lymphocytes. It is hypothesized that identification of CTL target schizont antigens will aid the development of a sub-unit vaccine. We exploited the availability of the complete genome sequence data and bioinformatics tools to identify genes encoding secreted or membrane anchored proteins that may be processed and presented by the MHC class I molecules of infected cells to CTL.

Results

Of the 986 predicted open reading frames (ORFs) encoded by chromosome 1 of the T. parva genome, 55 were selected based on the presence of a signal peptide and/or a transmembrane helix domain. Thirty six selected ORFs were successfully cloned into a eukaryotic expression vector, transiently transfected into immortalized bovine skin fibroblasts and screened in vitro using T. parva-specific CTL. Recognition of gene products by CTL was assessed using an IFN-γ ELISpot assay. A 525 base pair ORF encoding a 174 amino acid protein, designated Tp2, was identified by T. parva-specific CTL from 4 animals. These CTL recognized and lysed Tp2 transfected skin fibroblasts and recognized 4 distinct epitopes. Significantly, Tp2 specific CD8+ T cell responses were observed during the protective immune response against sporozoite challenge.

Conclusion

The identification of an antigen containing multiple CTL epitopes and its apparent immunodominance during a protective anti-parasite response makes Tp2 an attractive candidate for evaluation of its vaccine potential.