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Characterization of circulating T cells specific for tumor-associated antigens in melanoma patients

Nature Medicine volume 5, pages 677–685 (1999)Cite this article

Abstract

We identified circulating CD8+ T-cell populations specific for the tumor-associated antigens (TAAs) MART-1 (27-35) or tyrosinase (368-376) in six of eleven patients with metastatic melanoma using peptide/HLA-A*0201 tetramers. These TAA-specific populations were of two phenotypically distinct types: one, typical for memory/effector T cells; the other, a previously undescribed phenotype expressing both naive and effector cell markers. This latter type represented more than 2% of the total CD8+ T cells in one patient, permitting detailed phenotypic and functional analysis. Although these cells have many of the hallmarks of effector T cells, they were functionally unresponsive, unable to directly lyse melanoma target cells or produce cytokines in response to mitogens. In contrast, CD8+ T cells from the same patient were able to lyse EBV-pulsed target cells and showed robust allogeneic responses. Thus, the clonally expanded TAA-specific population seems to have been selectively rendered anergic in vivo. Peptide stimulation of the TAA-specific T-cell populations in other patients failed to induce substantial upregulation of CD69 expression, indicating that these cells may also have functional defects, leading to blunted activation responses. These data demonstrate that systemic TAA-specific T-cell responses can develop de novo in cancer patients, but that antigen-specific unresponsiveness may explain why such cells are unable to control tumor growth.

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Figure 1: Identification of TAA-specific CD8+ T cells by tetramer staining from PBMCs of six patients with metastatic melanoma.
Figure 2: Phenotypic analyses from ten-color FACS of PBMCs from patient 017.
Figure 3: Cytolytic activity of directly sorted and CD8-enriched T cells.
Figure 4: Intracellular cytokine staining of antigen-specific T cells.
Figure 5: Expression of CD69 by antigen-specific T cells after peptide stimulation.
Figure 6: Changes in antigen-specific CD8+ T-cell populations with chemotherapy in patient 017.

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Acknowledgements

We thank P. Roche (Mayo Clinic, Rochester, Minnesota) for the immunohistochemical staining of the tumor biopsy from patient 017; X.-S. He for suggesting the CD69 upregulation assay; and S. DeRosa, M. Mengozzi for discussions and J. Mumm and I. Tjioe for technical assistance. P.P.L. is supported by a physician-scientist award from the NIH (K08 CA72976). C.Y. is supported by the Burroughs Wellcome Fund Career Award and the NIH. P.A.S. is a Howard Hughes Medical Institute predoctoral fellow. M.R. is supported by NIH grants CA-42509 and CA-81543. This study was funded by grants from the Howard Hughes Medical Institute and the National Institutes of Health to M.M.D.

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Author notes

  1. P.P.L and C.Y. contributed equally to this study

Authors and Affiliations

  1. Howard Hughes Medical Institute/Department of Microbiology and Immunology, Stanford University, Stanford, 94305, California , USA

    Peter P. Lee, Peter A. Savage & Mark M. Davis

  2. Blood Center, Stanford University, Stanford , 94305, California, USA

    Lawrence Fong & Dirk Brockstedt

  3. Department of Surgery/Division of Surgical Oncology Stanford University, Stanford, 94305, California, USA

    Denise Johnson

  4. Department of Genetics, Stanford University, Stanford, 94305, California, USA

    Mario Roederer

  5. Department of Dermatology, Stanford University,

    Susan Swetter

  6. Palo Alto Veterans Administration Hospital,

    Susan Swetter

  7. Department of Medicine, University of Washington,

    Cassian Yee, John Thompson & Philip D. Greenberg

  8. Fred Hutchinson Cancer Research Center, Seattle, 98109, Washington, USA

    Cassian Yee, John Thompson & Philip D. Greenberg

  9. USC Norris Cancer Center, Los Angeles, 90033, California, USA

    Jeffrey S. Weber

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Correspondence to Mark M. Davis.

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Lee, P., Yee, C., Savage, P. et al. Characterization of circulating T cells specific for tumor-associated antigens in melanoma patients. Nat Med 5, 677–685 (1999). https://doi.org/10.1038/9525

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