Advances in the quantification of mitochondrial function in primary human immune
cells through extracellular flux analysis.
Authors Nicholas D, Proctor EA, Raval FM, Ip BC, Habib C, Ritou E, Grammatopoulos TN,
Steenkamp D, Dooms H, Apovian CM, Lauffenburger DA, Nikolajczyk BS
Submitted By Barbara Nikolajczyk on 4/3/2017
Status Published
Journal PLoS ONE
Year 2017
Date Published
Volume : Pages 12 : e0170975
PubMed Reference 28178278
Abstract Numerous studies show that mitochondrial energy generation determines the
effectiveness of immune responses. Furthermore, changes in mitochondrial
function may regulate lymphocyte function in inflammatory diseases like type 2
diabetes. Analysis of lymphocyte mitochondrial function has been facilitated by
introduction of 96-well format extracellular flux (XF96) analyzers, but the
technology remains imperfect for analysis of human lymphocytes. Limitations in
XF technology include the lack of practical protocols for analysis of archived
human cells, and inadequate data analysis tools that require manual quality
checks. Current analysis tools for XF outcomes are also unable to automatically
assess data quality and delete untenable data from the relatively high number of
biological replicates needed to power complex human cell studies. The objectives
of work presented herein are to test the impact of common cellular manipulations
on XF outcomes, and to develop and validate a new automated tool that
objectively analyzes a virtually unlimited number of samples to quantitate
mitochondrial function in immune cells. We present significant improvements on
previous XF analyses of primary human cells that will be absolutely essential to
test the prediction that changes in immune cell mitochondrial function and fuel
sources support immune dysfunction in chronic inflammatory diseases like type 2
diabetes.


Investigators with authorship
NameInstitution
Barbara NikolajczykUniversity of Kentucky

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