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In situ detection of virus- and tumor-specific T-cell immunity

Nature Medicine volume 6, pages 1056–1060 (2000)Cite this article

The development of strategies to visualize antigen-specific T-cell responses by flow cytometry has radically changed our understanding of the dynamics and magnitude of virus-induced T-cell immunity. Through the use of tetrameric–major histocompatibility complex (MHC) complexes, massive expansions of antigen-specific CD8+ T cells have been characterized for a variety of pathogens1,2,3,4,5. In addition, this technique has been used for the detection of auto-reactive and tumor-reactive T-cell immunity in vitiligo and melanoma patients6,7, demonstrating its value for the detection of low frequencies of self-reactive T cells. Here, we describe the use of MHC tetramers for the direct in situ visualization of antigen-specific T-cell responses. The combined use of MHC tetramer technology and confocal laser scanning microscopy (CLSM) allows detection of both virus- and tumor-specific CD8+ T cells in lymphoid organs, peripheral tissues and tumor infiltrates.

MHC tetramer staining of viable tissue sections

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Figure 1: Visualization and enumeration of antigen-specific T cells in TCR-transgenic mice.
Figure 2: In situ detection of polyclonal virus-specific CD8+ T cells.
Figure 3: Cell suspensions (3 × 106/ml) of lymph node cells from B10.F5 mice were incubated with APC-labeled NP366-tetramers for 50 min at the indicated temperature, followed by incubation with FITC-labeled anti-CD8b monoclonal antibody on ice.
Figure 4: In situ detection of tumor-specific CD8+ T cells in tumor tissue and lymph nodes.

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Acknowledgements

We thank P. Peters (NCI), M. Gonzalez-Fernandez and R. Noelle (Dartmouth Medical School) for their helpful suggestions, M. Toebes for MHC tetramers, M. de Rover, L. Bosman and K. Kits (Department of Neurophysiology, Free University of Amsterdam) for technical assistance in operating the microtome. This research was supported by the Dutch Cancer Society (grant NKI 99-2036).

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  1. Florry A. Vyth-Dreese and Ton N.M. Schumacher: T.N.M.S and F.A.V.-D. contributed equally to this study

Authors and Affiliations

  1. Department of Immunology, The Netherlands Cancer Institute, Plesmanlaan 121, Amsterdam, 1066 CX, The Netherlands

    John B.A.G. Haanen, Monique G.C.T. van Oijen, Felicia Tirion, Ada M. Kruisbeek, Florry A. Vyth-Dreese & Ton N.M. Schumacher

  2. Department of Cell Biology, The Netherlands Cancer Institute, Plesmanlaan 121, Amsterdam, 1066 CX, The Netherlands

    Lauran C.J.M. Oomen

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Correspondence to Ton N.M. Schumacher.

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Haanen, J., van Oijen, M., Tirion, F. et al. In situ detection of virus- and tumor-specific T-cell immunity. Nat Med 6, 1056–1060 (2000). https://doi.org/10.1038/79573

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