Flow cytometric visualisation of cytokine production by CD3-CD56+ NK cells and CD3+CD56+ NK-T cells in whole blood.
BACKGROUND: Natural killer (NK) cells produce multiple cytokines with potential immune regulatory roles. We standardised a whole-blood flow cytometry method to visualise intracellular cytokine production by NK cells for the study of NK cell biology and for clinical monitoring. METHODS: With a three-...
| Main Authors: | , , , , , , |
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| Format: | Journal article |
| Language: | English |
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2000
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| _version_ | 1826303240973582336 |
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| author | Mendes, R Bromelow, K Westby, M Galea-Lauri, J Smith, I O'Brien, M Souberbielle, B |
| author_facet | Mendes, R Bromelow, K Westby, M Galea-Lauri, J Smith, I O'Brien, M Souberbielle, B |
| author_sort | Mendes, R |
| collection | OXFORD |
| description | BACKGROUND: Natural killer (NK) cells produce multiple cytokines with potential immune regulatory roles. We standardised a whole-blood flow cytometry method to visualise intracellular cytokine production by NK cells for the study of NK cell biology and for clinical monitoring. METHODS: With a three-colour fluorescent labelling technique, specific cytokine production by NK or T cells was visualised directly in whole blood in the same sample after stimulation by phorbol 12-myristate 13-acetate (PMA) and ionomycin and by electronically gating on the CD3-ve/CD56+ve NK population or on the CD3+/CD56+ NK-T-cell population. RESULTS: Detectable levels of tumour necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma) but not of interleukin-2 (IL-2) or IL-4 were easily observed in NK cells. The visualisation of the cytokine production by NK cells was dependent on the addition of a Golgi transport inhibitor, Brefeldin A. Other known stimuli for NK cells (IL-2 and CD16 monoclonal antibody and incubation with K562, the NK-sensitive cell line) promoted IFN-gamma and TNF-alpha production in NK cells to a lesser extent than did PMA and ionomycin stimulation. CONCLUSIONS: This whole-blood flow cytometric assay appears to be an useful and easy method to examine cytokine production by NK cells and/or by CD3+CD56+ NK-T lymphocytes in patients with relevant diseases. |
| first_indexed | 2024-03-07T05:59:39Z |
| format | Journal article |
| id | oxford-uuid:ebbb8221-83a8-4b04-a9b1-46ce192be0fc |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T05:59:39Z |
| publishDate | 2000 |
| record_format | dspace |
| spelling | oxford-uuid:ebbb8221-83a8-4b04-a9b1-46ce192be0fc2022-03-27T11:12:03ZFlow cytometric visualisation of cytokine production by CD3-CD56+ NK cells and CD3+CD56+ NK-T cells in whole blood.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:ebbb8221-83a8-4b04-a9b1-46ce192be0fcEnglishSymplectic Elements at Oxford2000Mendes, RBromelow, KWestby, MGalea-Lauri, JSmith, IO'Brien, MSouberbielle, BBACKGROUND: Natural killer (NK) cells produce multiple cytokines with potential immune regulatory roles. We standardised a whole-blood flow cytometry method to visualise intracellular cytokine production by NK cells for the study of NK cell biology and for clinical monitoring. METHODS: With a three-colour fluorescent labelling technique, specific cytokine production by NK or T cells was visualised directly in whole blood in the same sample after stimulation by phorbol 12-myristate 13-acetate (PMA) and ionomycin and by electronically gating on the CD3-ve/CD56+ve NK population or on the CD3+/CD56+ NK-T-cell population. RESULTS: Detectable levels of tumour necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma) but not of interleukin-2 (IL-2) or IL-4 were easily observed in NK cells. The visualisation of the cytokine production by NK cells was dependent on the addition of a Golgi transport inhibitor, Brefeldin A. Other known stimuli for NK cells (IL-2 and CD16 monoclonal antibody and incubation with K562, the NK-sensitive cell line) promoted IFN-gamma and TNF-alpha production in NK cells to a lesser extent than did PMA and ionomycin stimulation. CONCLUSIONS: This whole-blood flow cytometric assay appears to be an useful and easy method to examine cytokine production by NK cells and/or by CD3+CD56+ NK-T lymphocytes in patients with relevant diseases. |
| spellingShingle | Mendes, R Bromelow, K Westby, M Galea-Lauri, J Smith, I O'Brien, M Souberbielle, B Flow cytometric visualisation of cytokine production by CD3-CD56+ NK cells and CD3+CD56+ NK-T cells in whole blood. |
| title | Flow cytometric visualisation of cytokine production by CD3-CD56+ NK cells and CD3+CD56+ NK-T cells in whole blood. |
| title_full | Flow cytometric visualisation of cytokine production by CD3-CD56+ NK cells and CD3+CD56+ NK-T cells in whole blood. |
| title_fullStr | Flow cytometric visualisation of cytokine production by CD3-CD56+ NK cells and CD3+CD56+ NK-T cells in whole blood. |
| title_full_unstemmed | Flow cytometric visualisation of cytokine production by CD3-CD56+ NK cells and CD3+CD56+ NK-T cells in whole blood. |
| title_short | Flow cytometric visualisation of cytokine production by CD3-CD56+ NK cells and CD3+CD56+ NK-T cells in whole blood. |
| title_sort | flow cytometric visualisation of cytokine production by cd3 cd56 nk cells and cd3 cd56 nk t cells in whole blood |
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