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Common Ewing sarcoma-associated antigens fail to induce natural T cell responses in both patients and healthy individuals

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Abstract

Disseminated or relapsed Ewing sarcoma (EwS) has remained fatal in the majority of patients. A promising approach to preventing relapse after conventional therapy is to establish tumor antigen-specific immune control. Efficient and specific T cell memory against the tumor depends on the expansion of rare T cells with native specificity against target antigens overexpressed by the tumor. Candidate antigens in EwS include six-transmembrane epithelial antigen of the prostate-1 (STEAP1), and the human cancer/testis antigens X-antigen family member 1 (XAGE1) and preferentially expressed antigen in melanoma (PRAME). Here, we screened normal donors and EwS patients for the presence of circulating T cells reactive with overlapping peptide libraries of these antigens by IFN-γ Elispot analysis. The majority of 22 healthy donors lacked detectable memory T cell responses against STEAP1, XAGE1 and PRAME. Moreover, ex vivo detection of T cells specific for these antigens in both blood and bone marrow were limited to a minority of EwS patients and required nonspecific T cell prestimulation. Cytotoxic T cells specific for the tumor-associated antigens were efficiently and reliably generated by in vitro priming using professional antigen-presenting cells and optimized cytokine stimulation; however, these T cells failed to interact with native antigen processed by target cells and with EwS cells expressing the antigen. We conclude that EwS-associated antigens fail to induce efficient T cell receptor (TCR)-mediated antitumor immune responses even under optimized conditions. Strategies based on TCR engineering could provide a more effective means to manipulating T cell immunity toward targeted elimination of tumor cells.

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Abbreviations

aAPC:

Artificial antigen-presenting cell

APC:

Antigen-presenting cell

BM:

Bone marrow

CTL:

Cytotoxic T lymphocyte

DC:

Dendritic cell

EwS:

Ewing sarcoma

HLA:

Human leukocyte antigen

IFN-γ:

Interferon-γ

IL:

Interleukin

MC:

Mononuclear cell

MHC:

Major histocompatibility complex

PB:

Peripheral blood

PD-1:

Programmed cell death 1

PepMix:

Peptide mixture

PRAME:

Preferentially expressed antigen in melanoma

RT-PCR:

Reverse transcriptase polymerase chain reaction

STEAP1:

Six-transmembrane epithelial antigen of the prostate-1

TCR:

T cell receptor

Treg cell:

T cell with regulatory phenotype

VZV-IE62:

Varicella-zoster virus immediate antigen 62

XAGE1:

X-antigen family member 1

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Acknowledgements

This work was supported by pilot funding by the University of Muenster Faculty of Medicine “Innovative Medizinische Forschung (IMF)” program (to Christiane Chen) and by grant Nr. 2010.016.1 from Wilhelm Sander-Stiftung (to Claudia Rossig).

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical standards

Approval for using tumor, BM and/or PB samples from patients and healthy donors was obtained from the University of Muenster Ethical Board, and informed consent was obtained in accordance with the 1964 Declaration of Helsinki and its later amendments.

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Authors and Affiliations

  1. Department of Pediatric Hematology and Oncology, University Children’s Hospital Muenster, Albert-Schweitzer Campus 1, 48149, Münster, Germany

    Bianca Altvater, Sareetha Kailayangiri, Nadine Theimann, Martina Ahlmann, Nicole Farwick, Christiane Chen, Sibylle Pscherer, Ilka Neumann, Gabriele Mrachatz, Heribert Juergens & Claudia Rossig

  2. Department of Internal Medicine A, Hematology and Oncology, University Hospital Muenster, Albert-Schweitzer Campus 1, 48149, Münster, Germany

    Anna Hansmeier

  3. Department of Orthopedic Surgery, University Hospital Muenster, Albert-Schweitzer Campus 1, 48149, Münster, Germany

    Jendrik Hardes & Georg Gosheger

  4. Cells-in-Motion Cluster of Excellence (EXC 1003 – CiM), University of Muenster, Münster, Germany

    Claudia Rossig

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  1. Bianca Altvater
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  2. Sareetha Kailayangiri
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  3. Nadine Theimann
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  7. Sibylle Pscherer
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  10. Anna Hansmeier
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  14. Claudia Rossig
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Correspondence to Claudia Rossig.

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Altvater, B., Kailayangiri, S., Theimann, N. et al. Common Ewing sarcoma-associated antigens fail to induce natural T cell responses in both patients and healthy individuals. Cancer Immunol Immunother 63, 1047–1060 (2014). https://doi.org/10.1007/s00262-014-1574-3

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