Cargando…
Impedance-based analysis of Natural Killer cell stimulation
The use of impedance-based label free cell analysis is increasingly popular and has many different applications. Here, we report that a real-time cell analyzer (RTCA) can be used to study the stimulation of Natural Killer (NK) cells. Engagement of NK cells via plate-bound antibodies directed against...
Autores principales: | , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2018
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5862859/ https://www.ncbi.nlm.nih.gov/pubmed/29563530 http://dx.doi.org/10.1038/s41598-018-23368-5 |
_version_ | 1783308296813281280 |
---|---|
author | Fasbender, Frank Watzl, Carsten |
author_facet | Fasbender, Frank Watzl, Carsten |
author_sort | Fasbender, Frank |
collection | PubMed |
description | The use of impedance-based label free cell analysis is increasingly popular and has many different applications. Here, we report that a real-time cell analyzer (RTCA) can be used to study the stimulation of Natural Killer (NK) cells. Engagement of NK cells via plate-bound antibodies directed against different activating surface receptors could be measured in real time using the label-free detection of impedance. The change in impedance was dependent on early signal transduction events in the NK cells as it was blocked by inhibitors of Src-family kinases and by inhibiting actin polymerization. While CD16 was the only receptor that could induce a strong change in impedance in primary NK cells, several activating receptors induced changes in impedance in expanded NK cells. Using PBMCs we could detect T cell receptor-mediated T cell activation and CD16-mediated NK cell activation in the same sample. Performing a dose-response analysis for the Src-family kinases inhibitor PP1 we show that T cells are more sensitive to inhibition compared to NK cells. Our data demonstrate that the RTCA can be used to detect physiological activation events in NK cells in a label-free and real-time fashion. |
format | Online Article Text |
id | pubmed-5862859 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-58628592018-03-27 Impedance-based analysis of Natural Killer cell stimulation Fasbender, Frank Watzl, Carsten Sci Rep Article The use of impedance-based label free cell analysis is increasingly popular and has many different applications. Here, we report that a real-time cell analyzer (RTCA) can be used to study the stimulation of Natural Killer (NK) cells. Engagement of NK cells via plate-bound antibodies directed against different activating surface receptors could be measured in real time using the label-free detection of impedance. The change in impedance was dependent on early signal transduction events in the NK cells as it was blocked by inhibitors of Src-family kinases and by inhibiting actin polymerization. While CD16 was the only receptor that could induce a strong change in impedance in primary NK cells, several activating receptors induced changes in impedance in expanded NK cells. Using PBMCs we could detect T cell receptor-mediated T cell activation and CD16-mediated NK cell activation in the same sample. Performing a dose-response analysis for the Src-family kinases inhibitor PP1 we show that T cells are more sensitive to inhibition compared to NK cells. Our data demonstrate that the RTCA can be used to detect physiological activation events in NK cells in a label-free and real-time fashion. Nature Publishing Group UK 2018-03-21 /pmc/articles/PMC5862859/ /pubmed/29563530 http://dx.doi.org/10.1038/s41598-018-23368-5 Text en © The Author(s) 2018 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/. |
spellingShingle | Article Fasbender, Frank Watzl, Carsten Impedance-based analysis of Natural Killer cell stimulation |
title | Impedance-based analysis of Natural Killer cell stimulation |
title_full | Impedance-based analysis of Natural Killer cell stimulation |
title_fullStr | Impedance-based analysis of Natural Killer cell stimulation |
title_full_unstemmed | Impedance-based analysis of Natural Killer cell stimulation |
title_short | Impedance-based analysis of Natural Killer cell stimulation |
title_sort | impedance-based analysis of natural killer cell stimulation |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5862859/ https://www.ncbi.nlm.nih.gov/pubmed/29563530 http://dx.doi.org/10.1038/s41598-018-23368-5 |
work_keys_str_mv | AT fasbenderfrank impedancebasedanalysisofnaturalkillercellstimulation AT watzlcarsten impedancebasedanalysisofnaturalkillercellstimulation |