Multiplexed division tracking dyes for proliferation-based clonal lineage tracing

Miles B. Horton; Giulio Prevedello; Julia M. Marchingo; Jie H.S. Zhou; Ken R. Duffy; Susanne Heinzel; Philip D. Hodgkin

doi:10.4049/jimmunol.1800481

Multiplexed division tracking dyes for proliferation-based clonal lineage tracing

Miles B. Horton, Giulio Prevedello, Julia M. Marchingo, Jie H.S. Zhou, Ken R. Duffy, Susanne Heinzel, Philip D. Hodgkin

Cell Signalling and Immunology

Research output: Contribution to journal › Article › peer-review

12 Citations (Scopus)

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.

Original language	English
Pages (from-to)	1097-1103
Number of pages	7
Journal	Journal of Immunology
Volume	201
Issue number	3
Early online date	23 Jul 2018
DOIs	https://doi.org/10.4049/jimmunol.1800481
Publication status	Published - 1 Aug 2018

ASJC Scopus subject areas

Immunology

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10.4049/jimmunol.1800481

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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.",

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AU - Heinzel, Susanne

AU - Hodgkin, Philip D.

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Multiplexed division tracking dyes for proliferation-based clonal lineage tracing

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