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Detection of virus-specific T cells and CD8+ T-cell epitopes by acquisition of peptide–HLA-GFP complexes: analysis of T-cell phenotype and function in chronic viral infections

Nature Medicine volume 9, pages 469–475 (2003)Cite this article

Abstract

Antigen-specific CD8+ T cells acquire peptide–major histocompatibility complex (MHC) clusters through T-cell receptor (TCR)–mediated endocytosis after specific antigen stimulation. We generated an antigen-presenting cell (APC) expressing human leukocyte antigen (HLA)-A*201 coupled to the enhanced green fluorescent protein (GFP), which delivered GFP to an antigen-specific T cell when pulsed with antigenic peptide. We quantitatively identified human T-cell lymphotropic virus type I (HTLV-I) Tax(11–19) peptide–specific T-cell populations in peripheral blood mononuclear cells (PBMCs) from patients with HTLV-I–associated neurologic disease and defined a new CD8+ T-cell epitope in the HTLV-I envelope region. Acquisition of peptide–HLA-GFP complexes by antigen-specific T cells could distinguish, with respect to phenotype and perforin production, T cells from the chronic viral infections cytomegalovirus and HTLV-I. This approach will be a powerful tool in understanding the role of antigen-specific T-cell responses in health and disease.

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Figure 1: Peptide-specific acquisition of peptide–HLA-GFP complexes by HTLV-I–specific CTL clones.
Figure 2: Quantitative detection of virus-specific CD8+ T cells from bulk PBMCs of HLA-A*201–positive HAM/TSP patients.
Figure 3: Detection of a new CD8+ T-cell epitope from the envelope region of HTLV-I.
Figure 4: Phenotypic and functional characterization of HTLV-I Tax(11–19)- and CMV pp65–specific T cells in PBMCs from HAM/TSP patients.

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Acknowledgements

We thank K. Yao, S. Gagnon and S. Soldan for helpful comments, and R. Turner for help with flow cytometry. U.T. was supported by the Japanese Society for the Promotion of Science and a US National Institutes of Health Research Fellowship.

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Author notes

  1. Utano Tomaru and Yoshihisa Yamano: U.T. and Y.Y. contributed equally to this work.

Authors and Affiliations

  1. Neuroimmunology Branch, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, Maryland, USA

    Utano Tomaru, Yoshihisa Yamano, William Biddison & Steven Jacobson

  2. Third Department of Internal Medicine, Faculty of Medicine, Kagoshima University, Kagoshima, Japan

    Masahiro Nagai

  3. Laboratory of Neurophysiology, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, Maryland, USA

    Dragan Maric

  4. Department of Molecular and Cellular Biochemistry, Ohio State University, Columbus, Ohio, USA

    Previn T.P. Kaumaya

Authors
  1. Utano Tomaru
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  2. Yoshihisa Yamano
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  7. Steven Jacobson
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Correspondence to Steven Jacobson.

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Tomaru, U., Yamano, Y., Nagai, M. et al. Detection of virus-specific T cells and CD8+ T-cell epitopes by acquisition of peptide–HLA-GFP complexes: analysis of T-cell phenotype and function in chronic viral infections. Nat Med 9, 469–475 (2003). https://doi.org/10.1038/nm845

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