The Journal of Experimental Medicine
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© The Rockefeller University Press, 0022-1007/1997/5/1815/ $5.00
The Journal of Experimental Medicine, Volume 185, Number 10, May 19, 1997 1815-1825

Articles

Kinetics and Extent of T Cell Activation as Measured with the Calcium Signal

Christoph Wülfing*, Joshua D. Rabinowitz{ddagger}, Craig Beeson{ddagger}, Michael D. Sjaastad§, Harden M. McConnell{ddagger}, and Mark M. Davis*,§

From the * Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, California 94305-5402; {ddagger} Department of Chemistry, Stanford University School of Medicine, Stanford, California 94305-5080; and the § Howard Hughes Medical Institute, Beckman Center, Stanford University, Stanford, California 94305-5428

We have characterized the calcium response of a peptide–major histocompatibility complex (MHC)-specific CD4+ T lymphocyte line at the single cell level using a variety of ligands, alone and in combination. We are able to distinguish four general patterns of intracellular calcium elevation, with only the most robust correlating with T cell proliferation. Whereas all three antagonist peptides tested reduce the calcium response to an agonist ligand, two give very different calcium release patterns and the third gives none at all, arguing that (a) antagonism does not require calcium release and (b) it involves interactions that are more T cell receptor proximal. We have also measured the time between the first T cell–antigen-presenting cell contact and the onset of the calcium signal. The duration of this delay correlates with the strength of the stimulus, with stronger stimuli giving a more rapid response. The dose dependence of this delay suggests that the rate-limiting step in triggering the calcium response is not the clustering of peptide–MHC complexes on the cell surface but more likely involves the accumulation of some intracellular molecule or complex with a half-life of a few minutes.

Address correspondence to M.M. Davis, Department of Microbiology and Immunology, HHMI, Beckman Center, Room B221, Stanford University, Stanford, California 94305-5428.

C. Wülfing was supported by an European Molecular Biology Organization long term fellowship, C. Beeson by the Cancer Research Institute, J.D. Rabinowitz by the Medical Scientist Training Program, and M.D. Sjaastad by the Howard Hughes Medical Institute. We also thank the Howard Hughes Medical Institute and the National Institutes of Health for supporting these studies.

1Abbreviations used in this paper: CHO, Chinese hamster ovary; MCC, moth cytochrome C; NF-AT, nuclear factor of activated T cell.


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