Simultaneous mRNA and protein quantification at the single-cell level delineates trajectories of CD4+ T-cell differentiation

Dominik Trzupek, Melanie Dunstan, Antony J. Cutler, Mercede Lee, Leila Godfrey, Dominik Aschenbrenner, Holm H. Uhlig, Linda S. Wicker, John A. Todd and Ricardo C. Ferreira

Aug 12, 2019
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Received Date: 26th July 19

The transcriptomic and proteomic characterisation of CD4+ T cells at the single-cell level has been performed traditionally by two largely exclusive types of technologies: single cell RNA-sequencing (scRNA-seq) technologies and antibody-based cytometry. Here we demonstrate that the simultaneous targeted quantification of mRNA and protein expression in single-cells provides a high-resolution map of human primary CD4+ T cells, and reveals precise trajectories of canonical T-cell lineage differentiation in blood and tissue. We report modest correlation between mRNA and protein in primary CD4+ T cells at the single-cell level, highlighting the importance of including quantitative protein expression data to characterise cell effector function. This approach provides a massively-parallel, cost-effective, solution to dissect the heterogeneity of immune cell populations and is ideally suited for the detailed immunophenotypic characterisation of rare and potentially pathogenic immune subsets. This transcriptomic and proteomic hybrid technology could have important clinical applications to re-define differentiation and activation of tissue-resident and blood human immune cells.

Read in full at bioRxiv.

This is an abstract of a preprint hosted on an independent third party site. It has not been peer reviewed but is currently under consideration at Nature Communications.

Nature Communications

Nature Research, Springer Nature