Horton, Miles B. and Prevedello, Giulio and Marchingo, J.M. and Zhou, Jie H.S. and Duffy, Ken R. and Heinzel, Susanne and Hodgkin, Philip D.
(2018)
Multiplexed Division Tracking Dyes for Proliferation-Based
Clonal Lineage Tracing.
Journal of Immunology, 201.
pp. 1097-1103.
ISSN 0022-1767
Abstract
The generation of cellular heterogeneity is an essential feature of immune responses. Understanding the heritability and asymmetry
of phenotypic changes throughout this process requires determination of clonal-level contributions to fate selection. Evaluating
intraclonal and interclonal heterogeneity and the influence of distinct fate determinants in large numbers of cell lineages, however,
is usually laborious, requiring familial tracing and fate mapping. In this study, we introduce a novel, accessible, high-throughput method
for measuring familial fate changes with accompanying statistical tools for testing hypotheses. The method combines multiplexing of
division tracking dyes with detection of phenotypic markers to reveal clonal lineage properties. We illustrate the method by studying
in vitro–activated mouse CD8+ T cell cultures, reporting division and phenotypic changes at the level of families. This approach has
broad utility as it is flexible and adaptable to many cell types and to modifications of in vitro, and potentially in vivo, fate monitoring
systems.
| Item Type: |
Article
|
| Additional Information: |
This work was supported by the National Health and Medical Research Council of
Australia via Project Grant 1010654, Program Grant 1054925, and a fellowship to
P.D.H., as well as Australian Government National Health and Medical Research
Council Independent Research Institutes Infrastructure Support Scheme Grant
361646. K.R.D. was supported by Science Foundation Ireland Grant 12IP1263.
J.M.M. was the recipient of a Sydney Parker Smith Postdoctoral Research Fellowship
from the Cancer Council of Victoria. J.H.S.Z. was the recipient of an Australian
Postgraduate Award. The research leading to these results has received funding from
the European Union Seventh Framework Programme (FP7/2007–2013) under Grant
Agreement 317040 (QuanTI) |
| Keywords: |
Multiplexed; Tracking Dyes; Proliferation-Based;
Clonal Lineage Tracing; |
| Academic Unit: |
Faculty of Science and Engineering > Research Institutes > Hamilton Institute |
| Item ID: |
13075 |
| Identification Number: |
https://doi.org/10.4049/jimmunol.1800481 |
| Depositing User: |
Dr Ken Duffy
|
| Date Deposited: |
19 Jun 2020 15:21 |
| Journal or Publication Title: |
Journal of Immunology |
| Publisher: |
American Association of Immunologists |
| Refereed: |
Yes |
| Funders: |
National Health and Medical Research Council of Australia, Science Foundation Ireland (SFI), Cancer Council of Victoria, European Union Seventh Framework Programme |
| URI: |
|
| Use Licence: |
This item is available under a Creative Commons Attribution Non Commercial Share Alike Licence (CC BY-NC-SA). Details of this licence are available
here |
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