Dynamic Interactions of Macrophages with T Cells during Antigen Presentation

doi:10.1084/jem.190.12.1909

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© The Rockefeller University Press, 0022-1007/1999/12/1909/ $5.00
The Journal of Experimental Medicine, Volume 190, Number 12, December 20, 1999 1909-1914

Brief Definitive Report

Dynamic Interactions of Macrophages with T Cells during Antigen Presentation

David M. Underhill^a, Michael Bassetti^a, Alexander Rudensky^a,b, and Alan Aderem^a
^a Department of Immunology, University of Washington, Seattle, Washington 98195
^b Howard Hughes Medical Institute, Seattle, Washington 98195

Correspondence to: Alan Aderem, Department of Immunology, University of Washington, H-574 Health Sciences, Box 357650, Seattle, WA 98195. Tel:206-616-5045 Fax:206-616-7237 E-mail:aaderem{at}u.washington.edu.

We have established a method for real-time video analysis ofthe interaction of antigen-presenting cells (APCs) with T cells.Green fluorescent protein expression controlled by a nuclearfactor of activated T cells (NFAT)-responsive promoter permitsthe visualization of productive antigen presentation in singleT cells. The readout is rapid (within 2 h) and semiquantitativeand allows analysis by video microscopy and flow cytometry.Using this approach, we demonstrate that macrophages have thecapacity to simultaneously activate multiple T cells. In addition,the interaction of T cells with macrophages is extraordinarilydynamic: after initial stable contact, the T cells migrate continuouslyon the surface of the macrophage and from APC to APC duringproductive antigen presentation. Thus, T cells sum up signalsfrom multiple interactions with macrophages during stimulation.

Key Words: nuclear factor of activated, T cells, hybridoma, green fluorescentprotein, video microscopy

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