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. 2018 Aug 29;20:191. doi: 10.1186/s13075-018-1692-y

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© The Author(s). 2018

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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 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.

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Fig. 2 — Representative gating strategy to identify CD8-positive and CD8-negative NK cell subsets. To increase parameter diversity detectable in a single seven-colour staining setup we have combined specifically expressed cell lineage markers that are labelled by the same fluorochrome, such as CD14-PE/CD56-PE/IgD-PE. Its cell-specific expression can be deconvoluted by sequential gating as exemplified by a: (left panel) gating on small cells (G1) with low granularity according to FSC/SSC characteristics allows exclusion of monocytes (CD14) and granulocytes (CD16); (middle panel) exclusion of B lymphocytes by CD19 and T lymphocytes by CD3 (G2); and (right panel) CD19/CD3-double negative cells were analysed for CD56 versus CD16 allowing us to identify NK cells by the co-expression of CD56 and CD16 (G3). b Classical NK cells were further analysed according to their expression of the CD8 receptor in five exemplary responders (R; top row) and non-responders (NR; bottom row)