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Reconciling Estimates of Cell Proliferation from Stable Isotope Labeling Experiments
- Source :
- PLoS Computational Biology, Vol 11, Iss 10, p e1004355 (2015), PLoS Computational Biology, 11(10). Public Library of Science, PLoS Computational Biology
- Publication Year :
- 2015
-
Abstract
- 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 2H2-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.<br />Author Summary Stable isotope labeling is used to quantify the rate at which living cells proliferate and die in humans. It has been central to a number of seminal studies, particularly in viral infections such as HIV-1, and leukemia. However, different labels (deuterated water or deuterated glucose) yield different estimates for the rate of cell proliferation and loss; this hampers our understanding and weakens our confidence in this important technique. We performed in vitro and in vivo experiments as well as a new analysis of existing data to directly compare the two labels. This reveals that a major source of the discrepancy lies in the difficulty of assessing deuterated glucose availability. We reconcile published studies and provide recommendations to avoid this problem in the future.
- Subjects :
- Normalization (statistics)
Radioisotope Dilution Technique
Lymphocyte
Context (language use)
Lymphocyte proliferation
Sensitivity and Specificity
Cellular and Molecular Neuroscience
In vivo
Genetics
medicine
Humans
Lymphocyte Count
Lymphocytes
Deuterium Oxide
Molecular Biology
lcsh:QH301-705.5
Ecology, Evolution, Behavior and Systematics
Cell Proliferation
Ecology
Chemistry
Cell growth
Reproducibility of Results
Deuterium
In vitro
3. Good health
Glucose
medicine.anatomical_structure
Computational Theory and Mathematics
Biochemistry
lcsh:Biology (General)
Isotope Labeling
Modeling and Simulation
Stable Isotope Labeling
Radiopharmaceuticals
Algorithms
Research Article
Subjects
Details
- Language :
- English
- ISSN :
- 1553734X and 15537358
- Volume :
- 11
- Issue :
- 10
- Database :
- OpenAIRE
- Journal :
- PLoS Computational Biology
- Accession number :
- edsair.doi.dedup.....7415c0ee4bfcc172383aa0f1f8dcddb5