Polyfunctional responses by human T cells result from sequential release of cytokines

Department of Chemical Engineering, Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Proceedings of the National Academy of Sciences (Impact Factor: 9.67). 12/2011; 109(5):1607-12. DOI: 10.1073/pnas.1117194109
Source: PubMed

ABSTRACT The release of cytokines by T cells defines a significant part of their functional activity in vivo, and their ability to produce multiple cytokines has been associated with beneficial immune responses. To date, time-integrated end-point measurements have obscured whether these polyfunctional states arise from the simultaneous or successive release of cytokines. Here, we used serial, time-dependent, single-cell analysis of primary human T cells to resolve the temporal dynamics of cytokine secretion from individual cells after activation ex vivo. We show that multifunctional, Th1-skewed cytokine responses (IFN-γ, IL-2, TNFα) are initiated asynchronously, but the ensuing dynamic trajectories of these responses evolve programmatically in a sequential manner. That is, cells predominantly release one of these cytokines at a time rather than maintain active secretion of multiple cytokines simultaneously. Furthermore, these dynamic trajectories are strongly associated with the various states of cell differentiation suggesting that transient programmatic activities of many individual T cells contribute to sustained, population-level responses. The trajectories of responses by single cells may also provide unique, time-dependent signatures for immune monitoring that are less compromised by the timing and duration of integrated measures.

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Available from: David A Hafler, Sep 29, 2015
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    • "Two elegant studies have demonstrated that CD8+ T cells have an astounding functional and phenotypic diversity [29], and that they can evolve with time and exhibit programmatic trajectories of cytokine secretion [30]. Among our new RSV-specific TCR Tg lines, we found that the “built-in” profile of each TCR clone was retained through a large range of peptide concentrations. "
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    ABSTRACT: Our previous work has characterized the functional and clonotypic features of two respiratory syncytial virus (RSV) epitope-specific T cell responses in mice. Following single-cell sequencing, we selected T cell receptor sequences to represent both a public and a private clone specific for the dominant KdM282-90 epitope for the generation of T cell receptor transgenic (TCR Tg) mice. We evaluated cells from these TCR Tg strains for three major functions of CD8+ T cells: proliferation, cytokine production and cytolytic activity. In vitro comparisons of the functional characteristics of T cells from the newly-generated mice demonstrated many similarities in their responsiveness to cognate antigen stimulation. Cells from both TRBV13-1 (private) and TRBV13-2 (public) TCR Tg mice had similar affinity, and proliferated similarly in vitro in response to cognate antigen stimulation. When transferred to BALB/c mice, cells from both strains demonstrated extensive proliferation in mediastinal lymph nodes following RSV infection, with TRBV13-2 demonstrating better in vivo proliferation. Both strains similarly expressed cytokines and chemokines following stimulation, and had similar Granzyme B and perforin expression, however cells expressing TRBV13-1 demonstrated better cytolytic activity than TRBV13-2 cells. These new, well-characterized mouse strains provide new opportunities to study molecular mechanisms that control the phenotype and function of CD8+ T cell responses.
    PLoS ONE 06/2014; 9(6):e99249. DOI:10.1371/journal.pone.0099249 · 3.23 Impact Factor
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    • "M1 and M2 macrophages, or Th1, Th2, and Th17 T-cells (Busse et al., 2010; Hong et al., 2011; Mendoza and Pardo, 2010; Santoni et al., 2008), which are usually defined by a specific set and relative amounts of secreted mediators, as well as by the set of mediators promoting their differentiation. In this context, single-cell experimental techniques may prove useful in refining cellular phenotype classification, by determining, e.g. if cytokine production by different phenotypes occurs in a sequential or simultaneous manner (Han et al., 2012). "
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    Mechanisms of ageing and development 01/2014; 136-137. DOI:10.1016/j.mad.2014.01.004 · 3.40 Impact Factor
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    • "However, the vast majority of unique TCR sequences are present at low frequencies, as would be expected from early estimates of possible TCR diversity34. Considering these data, estimates of TCRβ repertoire diversity in an individual are difficult to calculate , yet lower bounds for total TCRβ diversity in a healthy adult27,80 are ∼3–4 × 106 approximately consistent with previous estimates83. Single TCR chain deep sequencing studies have also shown that the clonal diversity of memory cells is greater than might be expected, with only a minority of memory T cell clones clearly expanded84. "
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