Advances in the Quantification of Mitochondrial Function in Primary Human Immune Cells Through Extracellular Flux Analysis

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2017 Feb 9

PMID 28178278

Citations 45

Authors

Dequina Nicholas

Elizabeth A Proctor

Forum M Raval

Blanche C Ip

Chloe Habib

Eleni Ritou

Tom N Grammatopoulos

Devin Steenkamp

Hans Dooms

Caroline M Apovian

Douglas A Lauffenburger

Barbara S Nikolajczyk

Affiliations

Soon will be listed here.

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.

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