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Despite the demonstrated efficacy and broad applicability of checkpoint blockade, the mechanisms by which it exerts its antitumor effects are incompletely understood. A recent article in Nature Medicine describes an ex vivo platform for assessing early responses to checkpoint blockade and the properties of tumor immune contexture in correlation to clinical responses., Competing Interests: Declaration of interests C.C.B. is a Bridge Scholar of the Parker Institute for Cancer Immunotherapy. N.B. is an extramural member of the Parker Institute for Cancer Immunotherapy; receives research funds from Regeneron, Harbor Biomedical, DC Prime, and Dragonfly Therapeutics; and is on the advisory boards of Neon Therapeutics, Novartis, Avidea, Boehringer Ingelheim, Rome Therapeutics, Roswell Park Comprehensive Cancer Center, BreakBio, Carisma Therapeutics, CureVac, Genotwin, BioNTech, Gilead Therapeutics, Tempest Therapeutics, and the Cancer Research Institute., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Determining the antigen specificities of the endogenous T-cell repertoire is important for screening naturally occurring or therapy-induced T-cell immunity and may help identify novel targets for T-cell-based therapies. Here, we describe a rapid, sensitive, and high-throughput protocol for expanding antigen-specific T cells from human peripheral blood mononuclear cells in vitro following peptide stimulation and detecting antigen-specific effector cytokine formation by flow cytometry. Our approach can be applied to examining specific T-cell subsets from various tissues. For complete details on the use and execution of this protocol, please refer to Roudko et al. (2020) and Cimen Bozkus et al. (2019)., Competing Interests: N.B. is an extramural member of the Parker Institute for Cancer Immunotherapy, receives research funds from 10.13039/100009857Regeneron, Harbor Biomedical, DC Prime, and Dragonfly Therapeutics, and is on the advisory boards of Neon Therapeutics, Novartis, Avidea, Boehringer Ingelheim, Rome Therapeutics, Roswell Park Comprehensive Cancer Center, BreakBio, Carisma Therapeutics, CureVac, Genotwin, BioNTech, Gilead Therapeutics, Tempest Therapeutics, and the Cancer Research Institute. C.C.B. is a Bridge Scholar of the Parker Institute for Cancer Immunotherapy. N.B. and C.C.B. have a patent application (application no. 17/290,128), which utilizes the T-cell immunogenicity assay protocol described here. A.B.B. is currently an employee of Bristol Myers Squibb. T.E. declares no interests., (© 2021 The Authors.)