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Infection and Immunity, February 2008, p. 685-694, Vol. 76, No. 2
0019-9567/08/$08.00+0     doi:10.1128/IAI.01244-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Characterization of the Fine Specificity of Bovine CD8 T-Cell Responses to Defined Antigens from the Protozoan Parasite Theileria parva{triangledown}

Simon P. Graham,1* Roger Pellé,1 Mat Yamage,1,{ddagger} Duncan M. Mwangi,1 Yoshikazu Honda,1,§ Ramadhan S. Mwakubambanya,1,¶ Etienne P. de Villiers,1 Evelyne Abuya,1 Elias Awino,1 James Gachanja,1 Ferdinand Mbwika,1 Anthony M. Muthiani,1 Cecelia Muriuki,1 John K. Nyanjui,1 Fredrick O. Onono,1,|| Julius Osaso,1 Victor Riitho,1 Rosemary M. Saya,1 Shirley A. Ellis,2 Declan J. McKeever,3,4,# Niall D. MacHugh,4 Sarah C. Gilbert,5 Jean-Christophe Audonnet,6 W. Ivan Morrison,4 Pierre van der Bruggen,7 and Evans L. N. Taracha1

International Livestock Research Institute, P.O. Box 30709, Nairobi 00100, Kenya,1 Immunology Division, Institute for Animal Health, Compton RG20 7NN,2 Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik, Midlothian EH26 0PZ,3 Division of Veterinary Clinical Sciences, Royal (Dick) School of Veterinary Studies, University of Edinburgh, Easter Bush, Roslin EH25 9RG,4 Wellcome Trust Centre for Human Genetics, Roosevelt Drive, Headington, Oxford OX3 7BN, United Kingdom,5 Discovery Research, Merial SAS, Lyon Gerland Laboratory, 254, rue Marcel Merieux, 69007 Lyon, France,6 Ludwig Institute for Cancer Research—Brussels Branch, Avenue Hippocrate 74, UCL 7459, B-1200 Brussels, Belgium7

Received 11 September 2007/ Returned for modification 15 October 2007/ Accepted 27 November 2007

Immunity against the bovine intracellular protozoan parasite Theileria parva has been shown to be mediated by CD8 T cells. Six antigens targeted by CD8 T cells from T. parva-immune cattle of different major histocompatibility complex (MHC) genotypes have been identified, raising the prospect of developing a subunit vaccine. To facilitate further dissection of the specificity of protective CD8 T-cell responses and to assist in the assessment of responses to vaccination, we set out to identify the epitopes recognized in these T. parva antigens and their MHC restriction elements. Nine epitopes in six T. parva antigens, together with their respective MHC restriction elements, were successfully identified. Five of the cytotoxic-T-lymphocyte epitopes were found to be restricted by products of previously described alleles, and four were restricted by four novel restriction elements. Analyses of CD8 T-cell responses to five of the epitopes in groups of cattle carrying the defined restriction elements and immunized with live parasites demonstrated that, with one exception, the epitopes were consistently recognized by animals of the respective genotypes. The analysis of responses was extended to animals immunized with multiple antigens delivered in separate vaccine constructs. Specific CD8 T-cell responses were detected in 19 of 24 immunized cattle. All responder cattle mounted responses specific for antigens for which they carried an identified restriction element. By contrast, only 8 of 19 responder cattle displayed a response to antigens for which they did not carry an identified restriction element. These data demonstrate that the identified antigens are inherently dominant in animals with the corresponding MHC genotypes.

* Corresponding author. Present address: Virology Department, Veterinary Laboratories Agency, Woodham Lane, New Haw, Addlestone, Surrey KT15 3NB, United Kingdom. Phone: 44 (0)1932 357 298. Fax: 44 (0)1932 357 239. E-mail: s.graham{at}vla.defra.gsi.gov.uk

{triangledown} Published ahead of print on 10 December 2007.

Editor: J. F. Urban, Jr.

{ddagger} Present address: World Organization for Animal Health (OIE), Sanseido Building, 4F, 2-4-10 Kojimachi, Chiyoda-ku, Tokyo 102-0083, Japan.

§ Present address: Vaxine Pty. Ltd., Department of Endocrinology and Diabetes, Flinders Medical Centre, Bedford Park, South Australia 5042, Australia.

Present address: Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology Hellas, P.O. Box 1527, Heraklion 711 10, Crete, Greece.

|| Present address: Department of Cardiology and Angiology, Hannover Medical School, Carl-Neuberg-Strasse 1, D-30625 Hannover, Germany.

# Present address: Royal Veterinary College, Hawkshead Lane, North Mymms, Hatfield, Hertfordshire AL9 7TA, United Kingdom.

Infection and Immunity, February 2008, p. 685-694, Vol. 76, No. 2
0019-9567/08/$08.00+0     doi:10.1128/IAI.01244-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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