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Reconciling Estimates of Cell Proliferation from Stable Isotope Labeling Experiments
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 r...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4593553/ https://www.ncbi.nlm.nih.gov/pubmed/26437372 http://dx.doi.org/10.1371/journal.pcbi.1004355 |
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author | Ahmed, Raya Westera, Liset Drylewicz, Julia Elemans, Marjet Zhang, Yan Kelly, Elizabeth Reljic, Rajko Tesselaar, Kiki de Boer, Rob J. Macallan, Derek C. Borghans, José A. M. Asquith, Becca |
author_facet | Ahmed, Raya Westera, Liset Drylewicz, Julia Elemans, Marjet Zhang, Yan Kelly, Elizabeth Reljic, Rajko Tesselaar, Kiki de Boer, Rob J. Macallan, Derek C. Borghans, José A. M. Asquith, Becca |
author_sort | Ahmed, Raya |
collection | PubMed |
description | 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)H(2)-glucose (D(2)-glucose) labeling studies consistently yield higher estimates of proliferation than deuterated water (D(2)O) 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. D(2)-glucose and D(2)O 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 D(2)-glucose and D(2)O. This showed that intrinsic differences between the two compounds do not cause differences in the proliferation rate estimates, but that estimates made using D(2)-glucose in vivo were susceptible to difficulties in normalization due to highly variable blood glucose enrichment. Analysis of three published human studies made using D(2)-glucose and D(2)O confirmed this problem, particularly in the case of short term D(2)-glucose labeling. Correcting for these inaccuracies in normalization decreased proliferation rate estimates made using D(2)-glucose and slightly increased estimates made using D(2)O; thus bringing the estimates from the two methods significantly closer and highlighting the importance of reliable normalization when using this technique. |
format | Online Article Text |
id | pubmed-4593553 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-45935532015-10-14 Reconciling Estimates of Cell Proliferation from Stable Isotope Labeling Experiments Ahmed, Raya Westera, Liset Drylewicz, Julia Elemans, Marjet Zhang, Yan Kelly, Elizabeth Reljic, Rajko Tesselaar, Kiki de Boer, Rob J. Macallan, Derek C. Borghans, José A. M. Asquith, Becca PLoS Comput Biol Research Article 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)H(2)-glucose (D(2)-glucose) labeling studies consistently yield higher estimates of proliferation than deuterated water (D(2)O) 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. D(2)-glucose and D(2)O 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 D(2)-glucose and D(2)O. This showed that intrinsic differences between the two compounds do not cause differences in the proliferation rate estimates, but that estimates made using D(2)-glucose in vivo were susceptible to difficulties in normalization due to highly variable blood glucose enrichment. Analysis of three published human studies made using D(2)-glucose and D(2)O confirmed this problem, particularly in the case of short term D(2)-glucose labeling. Correcting for these inaccuracies in normalization decreased proliferation rate estimates made using D(2)-glucose and slightly increased estimates made using D(2)O; thus bringing the estimates from the two methods significantly closer and highlighting the importance of reliable normalization when using this technique. Public Library of Science 2015-10-05 /pmc/articles/PMC4593553/ /pubmed/26437372 http://dx.doi.org/10.1371/journal.pcbi.1004355 Text en © 2015 Ahmed et al http://creativecommons.org/licenses/by/4.0/ 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 properly credited. |
spellingShingle | Research Article Ahmed, Raya Westera, Liset Drylewicz, Julia Elemans, Marjet Zhang, Yan Kelly, Elizabeth Reljic, Rajko Tesselaar, Kiki de Boer, Rob J. Macallan, Derek C. Borghans, José A. M. Asquith, Becca Reconciling Estimates of Cell Proliferation from Stable Isotope Labeling Experiments |
title | Reconciling Estimates of Cell Proliferation from Stable Isotope Labeling Experiments |
title_full | Reconciling Estimates of Cell Proliferation from Stable Isotope Labeling Experiments |
title_fullStr | Reconciling Estimates of Cell Proliferation from Stable Isotope Labeling Experiments |
title_full_unstemmed | Reconciling Estimates of Cell Proliferation from Stable Isotope Labeling Experiments |
title_short | Reconciling Estimates of Cell Proliferation from Stable Isotope Labeling Experiments |
title_sort | reconciling estimates of cell proliferation from stable isotope labeling experiments |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4593553/ https://www.ncbi.nlm.nih.gov/pubmed/26437372 http://dx.doi.org/10.1371/journal.pcbi.1004355 |
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