title: T cell epitopes of the timothy grass pollen allergen Phl p 5 of mice and men and the detection of allergen-specific T cells using Class II Ultimers.
abstract: BACKGROUND
  Knowledge of allergen-specific T cell epitopes is a prerequisite not only for therapeutic approaches but also for elucidating immunological mechanisms of type I allergy. Ex vivo detection of allergen-specific T cells using class II tetramer technology has become an important tool for investigating immune responses in atopic and healthy individuals.
  
  
  METHODS
  Using (3)H-thymidine incorporation assays, T cell epitopes specific for the major timothy grass pollen allergen Phl p 5.0101 were mapped in 11 allergic donors and two different mouse strains. Different protocols for expansion/restimulation of T cells from the blood of allergic donors and detection of allergen-specific T cells by Class II Ultimer staining were evaluated.
  
  
  RESULTS
  We identified several new Phl p 5.0101 class II T cell epitopes in allergic patients and confirmed previously published ones. Additionally, we discovered the major T cell epitopes in BALB/c and C57BL/6 mice. Using a novel Class II Ultimer, we detected epitope-specific T cells expanded from the blood of an allergic donor.
  
  
  CONCLUSIONS
  Epitope mapping of Phl p 5.0101 revealed an immunodominant epitope in BALB/c and C57BL/6 mice and an immunodominant region in humans (amino acids 259-282), which was recognized by 8 out of 11 allergic donors. Detection of Phl p 5-specific T cells was demonstrated using a Class II Ultimer specific for epitope 196-210. Successful detection of ultimer-positive T cells was strongly dependent on a resting phase after in vitro expansion.
authors.affiliation: Christian Doppler Laboratory for Allergy Diagnosis and Therapy, Department of Molecular Biology, University of Salzburg, Salzburg, Austria.
authors.forename: Angelika
authors.initials: A
authors.lastname: Stoecklinger
authors.forename: Sandra
authors.initials: S
authors.lastname: Scheiblhofer
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Roesler
authors.forename: Andreas
authors.initials: A
authors.lastname: Lang
authors.forename: Gerd
authors.initials: G
authors.lastname: Fastner
authors.forename: Felix
authors.initials: F
authors.lastname: Sedlmayer
authors.forename: Roland
authors.initials: R
authors.lastname: Lang
authors.forename: Martin
authors.initials: M
authors.lastname: Danzer
authors.forename: Josef
authors.initials: J
authors.lastname: Thalhamer
authors.forename: Richard
authors.initials: R
authors.lastname: Weiss
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D018604:Epitope Mapping; D018984:Epitopes, T-Lymphocyte / Q000276:immunology*; D005260:Female; D006801:Humans; D016056:Immunodominant Epitopes / Q000276:immunology; D007073:Immunoglobulin E / Q000097:blood / Q000276:immunology; D008213:Lymphocyte Activation / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000276:immunology*; D013194:Staining and Labeling; D013601:T-Lymphocytes / Q000276:immunology*
chemical_list: D018984:Epitopes, T-Lymphocyte; D016056:Immunodominant Epitopes; C076049:Phl p V protein, Phleum pratense; D010940:Plant Proteins; D007073:Immunoglobulin E