Mapping T cell epitopes by flow cytometry.
Epitope mapping by flow cytometry is a very modern approach that not only identifies T-cell epitopes but simultaneously allows for detailed analysis of the responding T-cell subsets including lineage, activation marker expression, and other markers of interest. The most frequently used approach is based on the identification of intracellular cytokines in secretion-inhibited activated T cells following stimulation with peptides or peptide pools. A more recently developed assay analyzes T-cell proliferation by measuring the decrease in carboxyfluorescein diacetate succinimidyl ester staining in proliferated cells. This article includes information on peptide configuration, a section on the design and efficient application of peptide pools, and working laboratory protocols for both assays.
Department of Medical Immunology, Charité, Humboldt Universität zu Berlin, Schumannstrasse 21, 10117 Berlin, Germany.
Bodo
B
Hoffmeister
Felix
F
Kiecker
Lydia
L
Tesfa
Hans-Dieter
HD
Volk
Louis J
LJ
Picker
Florian
F
Kern
D018604:Epitope Mapping / Q000379:methods*; D018984:Epitopes, T-Lymphocyte / Q000032:analysis*; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans
D018984:Epitopes, T-Lymphocyte