T-cell stimuli independently sum to regulate an inherited clonal division fate

J M Marchingo,A Kan,P D Hodgkin,G Prevedello,K R Duffy,S Heinzel

doi:10.1038/ncomms13540

Abstract

In the presence of antigen and costimulation, T cells undergo a characteristic response of expansion, cessation and contraction. Previous studies have revealed that population-level reproducibility is a consequence of multiple clones exhibiting considerable disparity in burst size, highlighting the requirement for single-cell information in understanding T-cell fate regulation. Here we show that individual T-cell clones resulting from controlled stimulation in vitro are strongly lineage imprinted with highly correlated expansion fates. Progeny from clonal families cease dividing in the same or adjacent generations, with inter-clonal variation producing burst-size diversity. The effects of costimulatory signals on individual clones sum together with stochastic independence; therefore, the net effect across multiple clones produces consistent, but heterogeneous population responses. These data demonstrate that substantial clonal heterogeneity arises through differences in experience of clonal progenitors, either through stochastic antigen interaction or by differences in initial receptor sensitivities.

Highlights

In the presence of antigen and costimulation, T cells undergo a characteristic response of expansion, cessation and contraction
Any clonal level answer to the question of relative concordance in division destiny (DD) must be reconciled with a further striking population level observation: T-cell DD is regulated by the type and the strength of the signals received, and many signal combinations result in both the means and variances of population DD distributions summing linearly, illustrated in Fig. 1c
We sought to identify the source of DD variation, and identify how signal integration that is additive at the population level results from, and is consistent with, clonal family behaviour

Summary

Introduction

In the presence of antigen and costimulation, T cells undergo a characteristic response of expansion, cessation and contraction. The net behaviour of these rare pathogen-specific clones dictates the characteristics of the population response and, for a given infection, results in a highly reproducible response magnitude Despite this consistency in population responses, in vivo measurements of clonal burst size and phenotype have revealed substantial heterogeneity between clones[2,3,4,5,6,7], highlighting the requirement for single-cell information in understanding T-cell fate regulation. To address these questions we develop and utilize a novel multiplex clonal division-tracking assay based on the combinatorial use of multiple division tracking dyes Using this technique we reveal that CD8 þ T-cell clones are imprinted with highly correlated division fates during the early immune response, such that progeny cells from clonal families cease dividing in the same or adjacent generations, with inter-clonal variation producing burst-size diversity. This clonal addition of signals results in reproducible population-level responses, with substantial clonal heterogeneity arising through differences in stochastic antigen interaction and initial receptor sensitivity

Methods

Results

Conclusion