Furthermore, there are challenges to obtaining sufficient numbers of functionally active NK cells from a patient’s blood because, despite being widespread throughout the body, NK cells represent just 2–18% of lymphocytes in human peripheral blood and it is challenging to obtain sufficient numbers of NK cells needed to overwhelm the number of tumor cells. While the number of target cells killed per effector varies by study, it is consistently noted that a minority of NK cells is responsible for the majority of killing events.
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Unfortunately for adoptive immunotherapies, many NK cells kill only once, with less than 30% capable of serial killing. Moreover, serial killer cells are often faster at delivering lytic hits and inducing target cell death than non-serial killer NK cells. Studies show that some cytotoxic immune cells, including T- and NK cells, are capable of killing multiple target cells, sequentially, in a process termed serial killing. Natural killer (NK) cells are appealing cells for immunotherapy because they are very potent effector lymphocytes of the innate immune system that can attack and kill many different tumor target cells without prior sensitization.
Red samples are stained CD16A and/or CD56. Light blue samples are the unstained control cells respective for each group. Expression of CD16A & CD56 by resting and IL-2 activated (LAK) bNK cells. NK cell staining for BV711 anti-CD16/CD56. CD244 is expressed by CD8+ T-cells (gated out with CD3) and all NK cells. NK cells were gated on as AF647 anti-CD244 positive cells. T-cells were gated out by Boolean not gate of FITC anti-CD3 positive T-cells. Live cells were gated by Boolean not gate, taking 7-AAD positive cells as dead cells. All cells were gated on by Pacific Blue anti-CD45. Single cells (without doublets) were gated by SSC height versus SSC width. Cells without debris were gated by forward scatter (FSC-A) v side scatter (SSC-A). First, beads were gated by high Pacific Orange signal.
Flow cytometry to determine the number of primary NK cells within peripheral blood mononuclear cells. The Creative Commons Public Domain Dedication waiver ( ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.Īdditional file 2: Figure S1. 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. 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. 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.
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