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Bioenergetic analysis of human peripheral blood mononuclear cells

N. Jones, J. Piasecka, A. H. Bryant, R. H. Jones, D. O. F. Skibinski, Nigel Francis, C. A. Thornton, David Skibinski Orcid Logo, Ruth Jones Orcid Logo, Cathy Thornton Orcid Logo, Aled Bryant Orcid Logo, Nick Jones Orcid Logo

Clinical & Experimental Immunology, Volume: 182, Issue: 1, Pages: 69 - 80

Swansea University Authors: Nigel Francis , David Skibinski Orcid Logo, Ruth Jones Orcid Logo, Cathy Thornton Orcid Logo, Aled Bryant Orcid Logo, Nick Jones Orcid Logo

DOI (Published version): 10.1111/cei.12662

Abstract

Leukocytes respond rapidly to pathogenic and other insults with responses ranging from cytokine production through to migration and phagocytosis. These are bioenergetically expensive and increased glycolytic flux provides ATP rapidly to support these essential functions. However, much of this work i...

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Published in: Clinical & Experimental Immunology
Published: 2015
URI: https://cronfa.swan.ac.uk/Record/cronfa23517
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These are bioenergetically expensive and increased glycolytic flux provides ATP rapidly to support these essential functions. However, much of this work is from animal studies. To better understand the relative role of glycolysis and oxidative phosphorylation in human leukocytes, especially their utility in a translational research setting, we undertook a study of human peripheral blood mononuclear cells (MNCs) bioenergetics. Glycolysis was essential during LPS-mediated IL-1&#xF062;, IL-6, and TNF&#xF061; production as 2-deoxy-D-glucose significantly decreased output of all three cytokines. After optimising cell numbers and the concentrations of all activators and inhibitors, oxidative phosphorylation and glycolysis profiles of fresh and cryopreserved/resuscitated MNCs were determined to explore the utility of MNCs for determining the bioenergetics health profile in multiple clinical settings. While the LPS-induced cytokine response did not differ significantly between fresh and resuscitated cells from the same donors, cryopreservation/resuscitation significantly affected some measures of oxidative phosphorylation mostly, but also glycolysis. Bioenergetics analysis of human MNCs provides a quick, effective means to measure the bioenergetics health index of many individuals but cryopreserved cells are not suitable for such an analysis. The translational utility of this approach was tested by comparing MNCs of pregnant and non-pregnant women to reveal increased bioenergetics health index with pregnancy but significantly reduced basal glycolysis and glycolytic capacity. 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spelling 2017-01-23T14:20:49.9550871 v2 23517 2015-09-30 Bioenergetic analysis of human peripheral blood mononuclear cells a726d297bbed7e5cd4c320f8f9dbf4d7 Nigel Francis Nigel Francis true false 328d16903f98c2b03a1cc64a7530322a 0000-0003-4077-6236 David Skibinski David Skibinski true false a1a281c8720685c422892ef168d4b279 0000-0001-5811-8827 Ruth Jones Ruth Jones true false c71a7a4be7361094d046d312202bce0c 0000-0002-5153-573X Cathy Thornton Cathy Thornton true false 021f7adc0923f6d2c2e2fc269758b8fe 0000-0002-4650-4672 Aled Bryant Aled Bryant true false 0fce0f7ddbdbfeb968f4e2f1e3f86744 0000-0003-4846-5117 Nick Jones Nick Jones true false 2015-09-30 FGMHL Leukocytes respond rapidly to pathogenic and other insults with responses ranging from cytokine production through to migration and phagocytosis. These are bioenergetically expensive and increased glycolytic flux provides ATP rapidly to support these essential functions. However, much of this work is from animal studies. To better understand the relative role of glycolysis and oxidative phosphorylation in human leukocytes, especially their utility in a translational research setting, we undertook a study of human peripheral blood mononuclear cells (MNCs) bioenergetics. Glycolysis was essential during LPS-mediated IL-1, IL-6, and TNF production as 2-deoxy-D-glucose significantly decreased output of all three cytokines. After optimising cell numbers and the concentrations of all activators and inhibitors, oxidative phosphorylation and glycolysis profiles of fresh and cryopreserved/resuscitated MNCs were determined to explore the utility of MNCs for determining the bioenergetics health profile in multiple clinical settings. While the LPS-induced cytokine response did not differ significantly between fresh and resuscitated cells from the same donors, cryopreservation/resuscitation significantly affected some measures of oxidative phosphorylation mostly, but also glycolysis. Bioenergetics analysis of human MNCs provides a quick, effective means to measure the bioenergetics health index of many individuals but cryopreserved cells are not suitable for such an analysis. The translational utility of this approach was tested by comparing MNCs of pregnant and non-pregnant women to reveal increased bioenergetics health index with pregnancy but significantly reduced basal glycolysis and glycolytic capacity. More detailed analysis of discrete leukocyte populations would be required to understand the relative roles of glycolysis and oxidative phosphorylation during inflammation and other immune responses. Journal Article Clinical & Experimental Immunology 182 1 69 80 31 12 2015 2015-12-31 10.1111/cei.12662 COLLEGE NANME Medicine, Health and Life Science - Faculty COLLEGE CODE FGMHL Swansea University 2017-01-23T14:20:49.9550871 2015-09-30T12:35:13.4112942 Swansea University Medical School Medicine N. Jones 1 J. Piasecka 2 A. H. Bryant 3 R. H. Jones 4 D. O. F. Skibinski 5 Nigel Francis 6 C. A. Thornton 7 David Skibinski 0000-0003-4077-6236 8 Ruth Jones 0000-0001-5811-8827 9 Cathy Thornton 0000-0002-5153-573X 10 Aled Bryant 0000-0002-4650-4672 11 Nick Jones 0000-0003-4846-5117 12 0023517-14012016094417.pdf JonesetalBioenergeticsofHumanPBMCsacceptedversion.pdf 2016-01-14T09:44:17.2200000 Output 856535 application/pdf Accepted Manuscript true 2016-07-31T00:00:00.0000000 true
title Bioenergetic analysis of human peripheral blood mononuclear cells
spellingShingle Bioenergetic analysis of human peripheral blood mononuclear cells
Nigel, Francis
David, Skibinski
Ruth, Jones
Cathy, Thornton
Aled, Bryant
Nick, Jones
title_short Bioenergetic analysis of human peripheral blood mononuclear cells
title_full Bioenergetic analysis of human peripheral blood mononuclear cells
title_fullStr Bioenergetic analysis of human peripheral blood mononuclear cells
title_full_unstemmed Bioenergetic analysis of human peripheral blood mononuclear cells
title_sort Bioenergetic analysis of human peripheral blood mononuclear cells
author_id_str_mv a726d297bbed7e5cd4c320f8f9dbf4d7
328d16903f98c2b03a1cc64a7530322a
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author_id_fullname_str_mv a726d297bbed7e5cd4c320f8f9dbf4d7_***_Nigel, Francis_***_
328d16903f98c2b03a1cc64a7530322a_***_David, Skibinski_***_0000-0003-4077-6236
a1a281c8720685c422892ef168d4b279_***_Ruth, Jones_***_0000-0001-5811-8827
c71a7a4be7361094d046d312202bce0c_***_Cathy, Thornton_***_0000-0002-5153-573X
021f7adc0923f6d2c2e2fc269758b8fe_***_Aled, Bryant_***_0000-0002-4650-4672
0fce0f7ddbdbfeb968f4e2f1e3f86744_***_Nick, Jones_***_0000-0003-4846-5117
author Nigel, Francis
David, Skibinski
Ruth, Jones
Cathy, Thornton
Aled, Bryant
Nick, Jones
author2 N. Jones
J. Piasecka
A. H. Bryant
R. H. Jones
D. O. F. Skibinski
Nigel Francis
C. A. Thornton
David Skibinski
Ruth Jones
Cathy Thornton
Aled Bryant
Nick Jones
format Journal article
container_title Clinical & Experimental Immunology
container_volume 182
container_issue 1
container_start_page 69
publishDate 2015
institution Swansea University
doi_str_mv 10.1111/cei.12662
college_str Swansea University Medical School
hierarchytype
hierarchy_top_id swanseauniversitymedicalschool
hierarchy_top_title Swansea University Medical School
hierarchy_parent_id swanseauniversitymedicalschool
hierarchy_parent_title Swansea University Medical School
department_str Medicine{{{_:::_}}}Swansea University Medical School{{{_:::_}}}Medicine
document_store_str 1
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description Leukocytes respond rapidly to pathogenic and other insults with responses ranging from cytokine production through to migration and phagocytosis. These are bioenergetically expensive and increased glycolytic flux provides ATP rapidly to support these essential functions. However, much of this work is from animal studies. To better understand the relative role of glycolysis and oxidative phosphorylation in human leukocytes, especially their utility in a translational research setting, we undertook a study of human peripheral blood mononuclear cells (MNCs) bioenergetics. Glycolysis was essential during LPS-mediated IL-1, IL-6, and TNF production as 2-deoxy-D-glucose significantly decreased output of all three cytokines. After optimising cell numbers and the concentrations of all activators and inhibitors, oxidative phosphorylation and glycolysis profiles of fresh and cryopreserved/resuscitated MNCs were determined to explore the utility of MNCs for determining the bioenergetics health profile in multiple clinical settings. While the LPS-induced cytokine response did not differ significantly between fresh and resuscitated cells from the same donors, cryopreservation/resuscitation significantly affected some measures of oxidative phosphorylation mostly, but also glycolysis. Bioenergetics analysis of human MNCs provides a quick, effective means to measure the bioenergetics health index of many individuals but cryopreserved cells are not suitable for such an analysis. The translational utility of this approach was tested by comparing MNCs of pregnant and non-pregnant women to reveal increased bioenergetics health index with pregnancy but significantly reduced basal glycolysis and glycolytic capacity. More detailed analysis of discrete leukocyte populations would be required to understand the relative roles of glycolysis and oxidative phosphorylation during inflammation and other immune responses.
published_date 2015-12-31T03:39:33Z
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