HLA class II peptide tetramers vs allergen-induced proliferation for identification of allergen-specific CD4 T cells
Van Hemelen D, Mahler V, Fischer G, Fae I, Reichl-Leb V, Pickl W, Jutel M, Smolinska S, Ebner C, Bohle B, Jahn-Schmid B (2015)
Publication Type: Journal article
Publication year: 2015
Journal
Book Volume: 70
Pages Range: 49-58
Journal Issue: 1
DOI: 10.1111/all.12524
Abstract
Fluorescence-labeled MHC class II/peptide tetramer complexes are considered as optimal tools to characterize allergen-specific CD4(+) T cells, but this technique is restricted to frequently expressed HLA class II molecules and knowledge of immunodominant epitopes. In contrast, allergen-stimulated proliferation assessed by CFSE dilution is less sophisticated and widely applicable. The major mugwort allergen, Art v 1, contains only one single, immunodominant, HLA-DR1-restricted epitope (Art v 125-36 ). Thus, essentially all Art v 1-reactive cells should be identified by a HLA-DRB1*01:01/Art v 119-36 tetramer.We compared specificity and sensitivity of tetramer(+) and allergen-induced proliferating (CFSE(lo) ) CD4(+) T cells by flow cytometry.The frequency of tetramer(+) CD4(+) T cells determined ex vivo in PBMC of mugwort-allergic individuals ranged from 0 to 0.029%. After 2-3 weeks of in vitro expansion, sufficient tetramer(+) T cells for phenotyping were detected in 83% of Art v 125-36 -reactive T-cell lines (TCL) from mugwort-allergic individuals, but not in TCL from healthy individuals. The tetramers defined bona fide Th2 cells. Notably, Art v 125-36 -reactive TCL depleted of tetramer(+) T cells still reacted to the peptide, and only 44% of Art v 125-36 -specific T-cell clones were detected by the tetramer. CFSE(lo) CD4(+) T cells contained only 0.3-10.7% of tetramer(+) T cells and very low proportions of Th2 cells.Allergen-specific T cells can be identified by HLA class II tetramers with high specificity, but unexpected low sensitivity. In contrast, allergen-stimulated CFSE(lo) CD4(+) T cells contain extremely high fractions of bystander cells. Therefore, for T-cell monitoring, either method should be interpreted with caution.
Authors with CRIS profile
Additional Organisation(s)
Involved external institutions
Medizinische Universität Wien
Austria (AT)
Wrocław Medical University / Uniwersytet Medyczny we Wrocławiu
Poland (PL)
Allergie-Ambulatorium Reumannplatz
Austria (AT)
Austria (AT)
Wrocław Medical University / Uniwersytet Medyczny we Wrocławiu
Poland (PL)
Allergie-Ambulatorium Reumannplatz
Austria (AT)
How to cite
APA:
Van Hemelen, D., Mahler, V., Fischer, G., Fae, I., Reichl-Leb, V., Pickl, W.,... Jahn-Schmid, B. (2015). HLA class II peptide tetramers vs allergen-induced proliferation for identification of allergen-specific CD4 T cells. Allergy, 70(1), 49-58. https://dx.doi.org/10.1111/all.12524
MLA:
Van Hemelen, D., et al. "HLA class II peptide tetramers vs allergen-induced proliferation for identification of allergen-specific CD4 T cells." Allergy 70.1 (2015): 49-58.
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