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. 2020 Apr 3;10:5885. doi: 10.1038/s41598-020-62244-z

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Identification of IFNγ response against peptide-reactive T cells using high-throughput imaging. (a) The 384-well screening system used to detect IFNγ-producing CD8⁺ T cells. (b) Detection of intracellular IFNγ levels in CD8⁺ T cells following peptide administration. PMA and ionomycin were used for non-specific T cell stimulation. Representative images are shown. (c) Bar graphs depicting the frequency of antigen-reactive CD8⁺ T cells. *P < 0.005, one-way analysis of variance, followed by Tukey’s multiple comparison test. (d) Peptide/A*11:01 MHC-tetramer staining for antigen-specific T cells analyzed by flow cytometry.