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. 2020 Apr 21;18:177. doi: 10.1186/s12967-020-02337-5

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© The Author(s) 2020

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 3 — Enrichment of JC-targeted T cells. Top: After stimulation with five non-overlapping JC virus peptide libraries (after stimulation, before selection: ORI), cells were stained with anti-CD45 (FITC), anti-CD3 (APC), anti-CD4 (APC/Cy7), anti-CD8 (PACBlue) and anti-IFNγ (PE), washed and resuspended in acquisition buffer containing 7-AAD. Note a minimal frequency of IFNγ + CD4 + and CD8 + T-cells, yet the absence of IFNγ-bright T-cells. Bottom: Final product (TARGET), after immunomagnetic selection of ORI with anti-IFNγ immunomagnetic beads. JC virus + T-cells are recognized as 7AAD-/CD45bright/SSClow/CD3 + cells with IFNγ on the surface. Enrichment of JC-virus specific T-cells to 65% among CD4 + and 68% among CD8 + T-cells is observed