The project is funded by The Danish Council for Independent research | Technology and Production 274-09-0281
Project period: July 2009 – December 2012.
Project participants:
Professor Gregers Jungersen, Project leader, DTU Vet
PhD student Lasse Eggers Pedersen, DTU Vet
Senior Scientist Ulla Riber, DTU Vet
Senior Scientist Morten Nielsen, DTU Systemsbiology
Post doc Dorte Rosenbek Fink, DTU Vet
Professor Søren Buus, Copenhagen University, Department of International Health, Immunology and Microbiology
Supervisory Scientist William T. Golde, United States Department of Agriculture, Agricultural Research Service, Plum Island Animal Disease Center
Danish Summary
Virusinfektioner i husdyr udgør en konstant velfærdsmæssig og økonomisk belastning verden over. Projektet vil levere nye højteknologiske reagenser til at måle immunsystemets dræberceller over for virusinficerede celler i svin. Dette vil være et ekstremt vigtigt værktøj i arbejdet med at udvikle nye effektive vacciner mod sygdomme som mund- og klovesyge og influenza.
Alle celler i kroppen viser på deres overflade små brudstykker af, hvad de indeholder. En celle angrebet af virus vil derfor fremvise virus brudstykker, der som en nøgle i en lås på dræbercellerne vil aktivere dem til at slå den angrebne celle ihjel. Herved standses virus i at dele sig, og den cellemedierede immunitet bliver således ofte det vigtigste forsvar mod virusinfektioner.
Virus-nøglen består af cellens egne vævstypemolekyler og et lille stykke virus-peptid (kæde af 8 - 11 aminosyrer). Vi vil fremstille selvlysende kunstige virus-nøgler (MHC klasse I tetramerer) til svin, så vi direkte kan farve og påvise dræberceller med aktivitet over for virus. Med tetramererne kan vi bestemme præcis, hvilke virus-peptider der giver det ønskede immunsvar og dermed også hvilke virus-komponenter, der kan anvendes i nye målrettede vacciner. Vi kan med tetramererne videre måle, om grisen har de ønskede dræberceller efter vaccination og vi vil lave computermodeller, der kan forudsige peptid-antigener i nye virus.
Projektgruppen består af forskere fra DTU og KU i samarbejde med førende veterinære virus og MHC forskere i USA.
Overall Objectives and Perspectives
Traditional vaccine technologies against virus infections employ live or dead virus particles to mimic the natural immune response seen after an infection. These vaccines are inefficient at inducing cell-mediated immune responses and are often associated with some degree of undesirable side-effects. Since viruses often change the immuno-dominant epitopes, both the natural and vaccine induced immunity may be inefficient against forthcoming infections. There is therefore a need for development of new vaccines targeting more conserved epitopes against virus infections in man and animals.
In this project we will develop tetramers of recombinant porcine peptide presenting molecules to directly identify virus specific immune cells with antigen recognition of the presented peptides. Tetramers will allow direct verification of peptide immune reactivity and provide a tremendously more detailed assessment of cell-mediated immune responses following infectious diseases or vaccination. In the current project we will develop porcine class I tetramers for staining of virus-specific CD8+ cytotoxic T cells in pigs, but in the long run we also aim for bovine class I reagents, and for class II tetramers so we can analyze all CD4+ and CD8+ T cell subsets of both species. Then we will have a rapid and refined assessment of vaccine performance without the need for viral challenge, and we can accelerate vaccine development tremendously.
Results dissemination
4th European Veterinary Immunology Workshop (EVIW), Edinburgh, Scotland, UK, 2012
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Lasse Eggers Pedersen (Oral presentation): From Viral genome to specific peptide epitopes - Methods for identifying porcine T cell epitopes based on in silico predictions, in vitro identification and ex vivo verification
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Lasse Eggers Pedersen et al (Poster): Swine Leukocyte Antigen (SLA) class I allele typing of Danish swine herds and the identification of commonly expressed haplotypes using sequence specific low- and high resolution primers
9th International Veterinary Immunology Symposium (IVIS), Tokyo, Japan, 2010
AAI Annual meeting i Baltimore, USA, 2010
Publications
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Lasse Eggers Pedersen, Dorte Rosenbek Fink, Gregers Jungersen. Swine Leukocyte Antigen (SLA) class I allele typing of Danish swine herds and identification of commonly expressed haplotypes using sequence specific low and high resolution primers. Manuscript submitted June 2012, Animal Genetics.
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Lasse Eggers Pedersen, Michael Rasmussen, Mikkel Harndahl, Morten Nielsen, Søren Buus, Gregers Jungersen. An analysis of affinity and stability in the identification of peptides bound by Swine Leukocyte Antigens (SLA) combining matrix- and NetMHCpan based peptide selection. Manuscript in preparation, Oct. 2012.
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Lasse Eggers Pedersen, Solvej Østergaard Breum, Charlotte Kristiane Hjulsager, Lars Erik Larsen, Gregers Jungersen. Identification of T cell epitopes by staining of cytotoxic T cells from animals previously immunized with Influenza. Manuscript in preparation, Oct. 2012.
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Thakur A, Pedersen LE, Jungersen G. 2012. Immune Markers and Correlates of Protection for Vaccine Induced Immune Responses. Vaccine 30: 4907-20.
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