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The Journal of Immunology, 2000, 165: 613-617.
Copyright © 2000 by The American Association of Immunologists


CUTTING EDGE

Cutting Edge: In Situ Tetramer Staining of Antigen-Specific T Cells in Tissues1

Pamela J. Skinner*, Mark A. Daniels{dagger}, Clint S. Schmidt{dagger}, Stephen C. Jameson{dagger} and Ashley T. Haase*,2

* Department of Microbiology and Great Lakes Center for AIDS Research, and {dagger} Center for Immunology and Department of Laboratory Medicine and Pathology, University of Minnesota Medical School, Minneapolis, MN 55455

Staining Ag-specific T cells with fluorescently labeled tetrameric MHC/peptide complexes has provided a powerful experimental approach to characterizing the immune response. In this report, we describe an extension of this method to directly visualize Ag-specific T cells in tissues. We successfully stained transgenic T cells with MHC tetramers in spleen sections from both 2C and OT-1 TCR transgenic mice. In addition, with the in situ tetramer staining technique, we detected a very small population of Ag-specific T cells in tissue after adoptive transfer of transgenic TCR T cells to a syngeneic nontransgenic mouse. We also show that the in situ tetramer technique can be applied to lightly fixed as well as frozen tissue, thus extending the method to archived tissue collections. This in situ tetramer staining technique offers a general approach to tracking the Ag-specific T cells in tissues.




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