Measurement of natural killer cell apoptotic inducing lytic proteins in peripheral blood mononuclear cells and isolated natural killer cells.
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1
Griffith University, Griffith Health Institute , Australia
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2
Bond University, Australia
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3
Queensland Health, Australia
Natural Killer (NK) cells are immune cells responsible for protecting the body from infection and malignancy. Cytotoxic NK cells contain secretory granules which store and release apoptotic inducing proteins such as perforin, granzyme A (GrzA) and granzyme B (GrzB). The aim of this study was to measure lytic proteins in samples of peripheral blood mononuclear cells (PBMCs) and isolated NK cells to determine if there is a difference in lytic protein expression. PBMCs from 19 healthy participants (mean age: 31±7.2SD) were isolated using density gradient centrifugation and negative isolation was used to separate the NK cells. Intracellular staining determined the content of perforin, GrzA and GrzB via flow cytometry in control and K562 stimulated samples. Statistical analysis included a repeated measures analysis of variance and significance was set at p<0.05. No significant changes were identified in lytic protein content in the control and K562 stimulated samples between the PBMCs and isolated NK cells. A significant increase in lytic proteins was observed in the isolated NK cells. The lymphocyte composition of the PBMC samples may account for this increase. NK cells only comprise of 15±8% of total circulating lymphocytes in the PBMC sample. Compared to the number of NK cells present in the isolated sample, there were fewer NK cells in the PBMC sample, translating to reduced levels of lytic proteins. Measurement of lytic proteins provides a direct measurement of NK cytotoxic activity and it may be beneficial for identifying cytotoxic activity deficiencies in the clinical setting.
Keywords:
Natural Killer,
cytotoxic activity,
Perforin,
GrzA,
GrzB.
Conference:
15th International Congress of Immunology (ICI), Milan, Italy, 22 Aug - 27 Aug, 2013.
Presentation Type:
Abstract
Topic:
Innate immunity
Citation:
Huth
TK,
Brenu
EW,
Ashton
KJ,
Staines
DR and
Gradisnik
SM
(2013). Measurement of natural killer cell apoptotic inducing lytic proteins in peripheral blood mononuclear cells and isolated natural killer cells..
Front. Immunol.
Conference Abstract:
15th International Congress of Immunology (ICI).
doi: 10.3389/conf.fimmu.2013.02.00940
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Received:
27 Jun 2013;
Published Online:
22 Aug 2013.
*
Correspondence:
Miss. Teilah K Huth, Griffith University, Griffith Health Institute, Gold Coast, Queensland, 4222, Australia, t.huth@griffith.edu.au