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Reconciling Estimates of Cell Proliferation from Stable Isotope Labeling Experiments.

Ahmed, R; Westera, L; Drylewicz, J; Elemans, M; Zhang, Y; Kelly, E; Reljic, R; Tesselaar, K; de Boer, RJ; Macallan, DC; et al. Ahmed, R; Westera, L; Drylewicz, J; Elemans, M; Zhang, Y; Kelly, E; Reljic, R; Tesselaar, K; de Boer, RJ; Macallan, DC; Borghans, JA; Asquith, B (2015) Reconciling Estimates of Cell Proliferation from Stable Isotope Labeling Experiments. PLoS Comput Biol, 11 (10). e1004355. ISSN 1553-7358
SGUL Authors: Macallan, Derek Clive Zhang, Yan

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Stable isotope labeling is the state of the art technique for in vivo quantification of lymphocyte kinetics in humans. It has been central to a number of seminal studies, particularly in the context of HIV-1 and leukemia. However, there is a significant discrepancy between lymphocyte proliferation rates estimated in different studies. Notably, deuterated (2)H2-glucose (D2-glucose) labeling studies consistently yield higher estimates of proliferation than deuterated water (D2O) labeling studies. This hampers our understanding of immune function and undermines our confidence in this important technique. Whether these differences are caused by fundamental biochemical differences between the two compounds and/or by methodological differences in the studies is unknown. D2-glucose and D2O labeling experiments have never been performed by the same group under the same experimental conditions; consequently a direct comparison of these two techniques has not been possible. We sought to address this problem. We performed both in vitro and murine in vivo labeling experiments using identical protocols with both D2-glucose and D2O. This showed that intrinsic differences between the two compounds do not cause differences in the proliferation rate estimates, but that estimates made using D2-glucose in vivo were susceptible to difficulties in normalization due to highly variable blood glucose enrichment. Analysis of three published human studies made using D2-glucose and D2O confirmed this problem, particularly in the case of short term D2-glucose labeling. Correcting for these inaccuracies in normalization decreased proliferation rate estimates made using D2-glucose and slightly increased estimates made using D2O; thus bringing the estimates from the two methods significantly closer and highlighting the importance of reliable normalization when using this technique.

Item Type: Article
Additional Information: © 2015 Ahmed et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Keywords: Algorithms, Cell Proliferation, Deuterium, Deuterium Oxide, Glucose, Humans, Isotope Labeling, Lymphocyte Count, Lymphocytes, Radioisotope Dilution Technique, Radiopharmaceuticals, Reproducibility of Results, Sensitivity and Specificity, Lymphocytes, Humans, Deuterium, Deuterium Oxide, Glucose, Radiopharmaceuticals, Radioisotope Dilution Technique, Lymphocyte Count, Sensitivity and Specificity, Reproducibility of Results, Isotope Labeling, Cell Proliferation, Algorithms, Bioinformatics, 06 Biological Sciences, 08 Information And Computing Sciences, 01 Mathematical Sciences
SGUL Research Institute / Research Centre: Academic Structure > Infection and Immunity Research Institute (INII)
Journal or Publication Title: PLoS Comput Biol
ISSN: 1553-7358
Language: eng
5 October 2015Published
26 May 2015Accepted
Publisher License: Creative Commons: Attribution 4.0
Project IDFunderFunder ID
103865Wellcome TrustUNSPECIFIED
G1001052Medical Research CouncilUNSPECIFIED
J007439Medical Research CouncilUNSPECIFIED
MR/J007439/1Medical Research CouncilUNSPECIFIED
PubMed ID: 26437372
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