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PNAS 109 (5): 1607-1612

Copyright © 2012 by the National Academy of Sciences.

From the Cover


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

Qing Hana,1, Neda Bagherib,1, Elizabeth M. Bradshawc, David A. Haflerd,e, Douglas A. Lauffenburgerb,e, and J. Christopher Lovea,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

Edited by Robert L. Coffman, Dynavax Technologies, Berkeley, CA, and approved November 14, 2011 (received for review October 18, 2011)

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-{gamma}, 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

Author contributions: Q.H., N.B., and J.C.L. designed research; Q.H., N.B., and E.M.B. performed research; Q.H., N.B., D.A.L., and J.C.L. analyzed data; and Q.H., N.B., D.A.H., D.A.L., and J.C.L. wrote the paper.

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

See Commentary on page 1359.

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

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