From the Cover

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

Qing Han^a,1, Neda Bagheri^b,1, Elizabeth M. Bradshaw^c, David A. Hafler^d,e, Douglas A. Lauffenburger^b,e, and J. Christopher Love^a,e,2

^aDepartment of Chemical Engineering, Koch Institute for Integrative Cancer Research, and ^bDepartment of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139; ^c Brigham and Women's Hospital, Boston, MA 02115; ^dDepartment of Neurology and Immunobiology, Yale University, New Haven, CT 06520; and ^e The Eli and Edythe L. Broad Institute, Cambridge, MA 02142

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-, 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.

Key Words: microengraving • multifunctionality • dynamical systems • computational biology

¹Q.H. and N.B. contributed equally to this work.

The authors declare no conflict of interest.

This article is a PNAS Direct Submission.

This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.1073/pnas.1117194109/-/DCSupplemental.

See Commentary on page 1359.

²To whom correspondence should be addressed. E-mail: clove{at}mit.edu.

Dynamic single-cell measurements of gene expression in primary lymphocytes: challenges, tools and prospects.

M. Polonsky, I. Zaretsky, and N. Friedman (2013)
Briefings in Functional Genomics 12, 99-108
 |  Abstract »  |  Full Text »  |  PDF »

Temporal Dynamics of the Primary Human T Cell Response to Yellow Fever Virus 17D As It Matures from an Effector- to a Memory-Type Response.

K. Blom, M. Braun, M. A. Ivarsson, V. D. Gonzalez, K. Falconer, M. Moll, H.-G. Ljunggren, J. Michaelsson, and J. K. Sandberg (2013)
J. Immunol. 190, 2150-2158
 |  Abstract »  |  Full Text »  |  PDF »

Construction and Evaluation of a Novel Recombinant T Cell Epitope-Based Vaccine against Coccidioidomycosis.

B. J. Hurtgen, C.-Y. Hung, G. R. Ostroff, S. M. Levitz, and G. T. Cole (2012)
Infect. Immun. 80, 3960-3974
 |  Abstract »  |  Full Text »  |  PDF »

TWEAK and cIAP1 Regulate Myoblast Fusion Through the Noncanonical NF-{kappa}B Signaling Pathway.

E. K. Enwere, J. Holbrook, R. Lejmi-Mrad, J. Vineham, K. Timusk, B. Sivaraj, M. Isaac, D. Uehling, R. Al-awar, E. LaCasse, et al. (2012)
Science Signaling 5, ra75
 |  Abstract »  |  Full Text »  |  PDF »

Travels in time: Assessing the functional complexity of T cells.

W. N. Haining (2012)
PNAS 109, 1359-1360
 |  Full Text »  |  PDF »