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.
Hoffmeister
Bodo
B
Department of Medical Immunology, Charité, Humboldt Universität zu Berlin, Schumannstrasse 21, 10117 Berlin, Germany.
Kiecker
Felix
F
Tesfa
Lydia
L
Volk
Hans-Dieter
HD
Picker
Louis J
LJ
Kern
Florian
F
D018604:Epitope Mapping / Q000379:methods*; D018984:Epitopes, T-Lymphocyte / Q000032:analysis*; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans
D018984:Epitopes, T-Lymphocyte