Estimating average cellular turnover from 5–bromo–2'–deoxyuridine (BrdU) measurements

Boer, Rob J. de; Mohri, Hiroshi; Ho, David D.; Perelson, Alan S.

doi:10.1098/rspb.2002.2316

Cited by 37 publications

(82 citation statements)

References 36 publications

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“…Division and death are normally quantified by following the accumulation and loss of cells labeled in vivo with BrdU or deuterium from heavy water or deuterated glucose, taken up by dividing cells during administration of label and diluted following its withdrawal (10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20). These experiments are typically performed over days to weeks and collectively have revealed that cell populations initially assumed to be homogenous may in fact comprise multiple subpopulations dividing and dying at different rates (kinetic heterogeneity) and/or that cells that are quiescent or have recently divided may have different susceptibilities to death (temporal heterogeneity).…”

mentioning

confidence: 99%

Temporal fate mapping reveals age-linked heterogeneity in naive T lymphocytes in mice

Hogan

Gossel

Yates

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

111

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Understanding how our T-cell compartments are maintained requires knowledge of their population dynamics, which are typically quantified over days to weeks using the administration of labels incorporated into the DNA of dividing cells. These studies present snapshots of homeostatic dynamics and have suggested that lymphocyte populations are heterogeneous with respect to rates of division and/or death, although resolving the details of such heterogeneity is problematic. Here we present a method of studying the population dynamics of T cells in mice over timescales of months to years that reveals heterogeneity in rates of division and death with respect to the age of the host at the time of thymic export. We use the transplant conditioning drug busulfan to ablate hematopoetic stem cells in young mice but leave the peripheral lymphocyte compartments intact. Following their reconstitution with congenically labeled (donor) bone marrow, we followed the dilution of peripheral host T cells by donor-derived lymphocytes for a year after treatment. Describing these kinetics with mathematical models, we estimate rates of thymic production, division and death of naive CD4 and CD8 T cells. Population-averaged estimates of mean lifetimes are consistent with earlier studies, but we find the strongest support for a model in which both naive T-cell pools contain kinetically distinct subpopulations of older host-derived cells with self-renewing capacity that are resistant to displacement by naive donor lymphocytes. We speculate that these incumbent cells are conditioned or selected for increased fitness through homeostatic expansion into the lymphopenic neonatal environment.T-cell homeostasis | mathematical modeling | Ki67 | kinetic heterogeneity N ormal adaptive immunity depends on maintaining populations of naive CD4 and CD8 T cells of sufficient sizes and diversities of antigen receptors. Mature naive cells are generated by the thymus and, once in the periphery, divide slowly and are lost either to death or differentiation into effector cells. It is known qualitatively how both cytokine (1-7) and T-cell receptor (TCR) (4, 8, 9) signals influence their survival and self-renewal through division, but we still lack a quantitative understanding of the rules that govern the development and persistence of our naive T-cell repertoires.To develop our understanding of lymphocyte homeostasis, much effort has been directed at defining the kinetics of T cells under normal physiological conditions. Division and death are normally quantified by following the accumulation and loss of cells labeled in vivo with BrdU or deuterium from heavy water or deuterated glucose, taken up by dividing cells during administration of label and diluted following its withdrawal (10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20). These experiments are typically performed over days to weeks and collectively have revealed that cell populations initially assumed to be homogenous may in fact comprise multiple subpopulations dividing and dying at different rates (kinetic ...

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mentioning

confidence: 99%

Temporal fate mapping reveals age-linked heterogeneity in naive T lymphocytes in mice

Hogan

Gossel

Yates

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

111

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“…Representative fits of our novel model to the data on BrdU labelling of CD3 þ CD45RA 2 CD4 þ and CD4 2 memory T cells in one monkey from each group are depicted in figure 3. The new model describes these data at least as well as previous models did [13,26,27]. Thanks to the kinetic heterogeneity, the model readily accounts for the biphasic down-slopes in the monkey H1284 with a high viral load.…”

Section: Kinetic Heterogeneitymentioning

confidence: 63%

“…Thus, if the first division is sufficient to breach the detection limit, the up-slope equation (2.4) is identical to that of equation (1.1) in §1 (which is an expected result). The de-labelling phase starts at T ¼ 10 days and we see that the down-slope depends on the detection limit, and hence does not reflect p, d, or p 2 d, as was derived in previous models [13,[25][26][27]. Importantly, the new model allows for a decline in the fraction of BrdU þ cells in self-renewing populations, in the absence of additional assumptions such as a source of unlabelled cells [13,[25][26][27] or a faster death rate of recently divided cells [28,29].…”

Section: Homogeneous Populationmentioning

confidence: 81%

“…To solve this problem, different authors have proposed different solutions. Several authors [13,[25][26][27] allowed for an external source of cells, for example coming from the thymus or from a compartment of quiescent cells, and by allowing the generation of unlabelled cells during the de-labelling phase they were able to explain the observed down-slopes. Others [28][29][30] argued that labelled cells have recently divided, and that recently divided cells should have a faster death rate than non-divided unlabelled cells, which also allows for a decline of the fraction of BrdU þ cells.…”

Section: Introductionmentioning

confidence: 99%

“…Thanks to the kinetic heterogeneity, the model readily accounts for the biphasic down-slopes in the monkey H1284 with a high viral load. Summarizing, we can account for the decline in the fraction of BrdU þ cells during de-labelling without having to invoke an external source of unlabelled cells [13,26,27] or a faster death rate of recently divided cells [28,29].…”

Section: Kinetic Heterogeneitymentioning

confidence: 99%

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A mechanistic model for bromodeoxyuridine dilution naturally explains labelling data of self-renewing T cell populations

Ganusov

Boer²

2013

J. R. Soc. Interface.

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Bromodeoxyuridine (BrdU) is widely used in immunology to detect cell division, and several mathematical models have been proposed to estimate proliferation and death rates of lymphocytes from BrdU labelling and delabelling curves. One problem in interpreting BrdU data is explaining the de-labelling curves. Because shortly after label withdrawal, BrdU þ cells are expected to divide into BrdU þ daughter cells, one would expect a flat down-slope. As for many cell types, the fraction of BrdU þ cells decreases during de-labelling, previous mathematical models had to make debatable assumptions to be able to account for the data. We develop a mechanistic model tracking the number of divisions that each cell has undergone in the presence and absence of BrdU, and allow cells to accumulate and dilute their BrdU content. From the same mechanistic model, one can naturally derive expressions for the mean BrdU content (MBC) of all cells, or the MBC of the BrdU þ subset, which is related to the mean fluorescence intensity of BrdU that can be measured in experiments. The model is extended to include subpopulations with different rates of division and death (i.e. kinetic heterogeneity). We fit the extended model to previously published BrdU data from memory T lymphocytes in simian immunodeficiency virusinfected and uninfected macaques, and find that the model describes the data with at least the same quality as previous models. Because the same model predicts a modest decline in the MBC of BrdU þ cells, which is consistent with experimental observations, BrdU dilution seems a natural explanation for the observed down-slopes in self-renewing populations.

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Reduced mitotic activity at the periphery of human embryonic stem cell colonies cultured in vitro with mitotically‐inactivated murine embryonic fibroblast feeder cells

Heng

Cao

Liu

et al. 2005

Cell Biochemistry & Function

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This study attempted to investigate whether different levels of mitotic activity exist within different physical regions of a human embryonic stem (hES) cell colony. Incorporation of 5-bromo-2-deoxyuridine (BrdU) within newly-synthesized DNA, followed by immunocytochemical staining was used as a means of detecting mitotically-active cells within hES colonies. The results showed rather surprisingly that the highest levels of mitotic activity are primarily concentrated within the central regions of hES colonies, whereas the peripheral regions exhibited reduced levels of cellular proliferation. Two hypothetical mechanisms are therefore proposed for hES colony growth and expansion. Firstly, it is envisaged that the less mitotically-active hES cells at the periphery of the colony are continually migrating outwards, thereby providing space for newly-divided daughter cells within the more mitotically-active central region of the hES colony. Secondly, it is proposed that the newly-divided hES cells within the central region of the colony somehow migrate to the outer periphery. This could possibly explain why the periphery of hES colonies are less mitotically-active, since there would obviously be an extended time-lag before newly-divided daughter cells are ready again for the next cell division. Further investigations need to be carried out to characterize the atypical mechanisms by which hES colonies grow and expand in size.

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Estimating average cellular turnover from 5–bromo–2'–deoxyuridine (BrdU) measurements

Cited by 37 publications

References 36 publications

Temporal fate mapping reveals age-linked heterogeneity in naive T lymphocytes in mice

Temporal fate mapping reveals age-linked heterogeneity in naive T lymphocytes in mice

A mechanistic model for bromodeoxyuridine dilution naturally explains labelling data of self-renewing T cell populations

Reduced mitotic activity at the periphery of human embryonic stem cell colonies cultured in vitro with mitotically‐inactivated murine embryonic fibroblast feeder cells

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