TY - DATA T1 - Supplementary Material for: 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 PY - 2012/04/03 AU - Stoecklinger A. AU - Scheiblhofer S. AU - Roesler E. AU - Lang A. AU - Fastner G. AU - Sedlmayer F. AU - Lang R. AU - Danzer M. AU - Thalhamer J. AU - Weiss R. UR - https://karger.figshare.com/articles/dataset/Supplementary_Material_for_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/5122975 DO - 10.6084/m9.figshare.5122975.v1 L4 - https://ndownloader.figshare.com/files/8708356 KW - C57BL/6 mouse KW - Phl p 5 KW - Epitope mapping KW - Tetramers KW - BALB/c mouse N2 - 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 3H-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. ER -