Novel fluorescence assay for tracking molecular and cellular allergen-protein interactions.
T cells recognizing nickel (Ni) are key mediators in human Ni allergy, which represents the most common form of human contact hypersensitivity. In contrast to well-characterized Ni-specific human T cell clones, molecular knowledge about the extra- and intracellular route(s) of antigen/allergen presentation and processing of Ni-specific epitopes is still fragmentary. Here, we demonstrate a new metal-specific fluorescent technique to detect and quantify metal ions, like Ni(2+), while they are associated with isolated metalloproteins. Moreover, utilizing the fluorescent metal sensor molecule Newport Green (NPG) a novel method has been developed, which permits the metal-specific detection of Ni(2+) binding to surface or intracellular structures of individual human antigen presenting cells by flow cytometry. We expect such metal-specific fluorescent analyses to contribute to a better basic understanding of molecular and cellular immune processes involved in Ni-specific T cell epitope generation and the pathogenesis of human nickel allergy.
Thierse
Hermann-Josef
HJ
Research Group Immunology and Proteomics, Department of Dermatology, University Medical Center Mannheim, University of Heidelberg, Mannheim, Germany. hermann-josef.thierse@haut.ma.uni-heidelberg.de
Helm
Stefanie
S
Pink
Matthias
M
Weltzien
Hans Ulrich
HU
D000485:Allergens; D000938:Antigen-Presenting Cells; D001402:B-Lymphocytes / Q000276:immunology*; D002460:Cell Line; D005434:Flow Cytometry / Q000379:methods*; D005455:Fluorescent Antibody Technique / Q000379:methods*; D005456:Fluorescent Dyes; D005470:Fluorometry; D006801:Humans; D008667:Metalloproteins / Q000378:metabolism*; D009000:Monocytes; D009532:Nickel / Q000032:analysis* / Q000276:immunology / Q000378:metabolism
D000485:Allergens; D005456:Fluorescent Dyes; D008667:Metalloproteins; D009532:Nickel