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Identification and translational validation of novel mammaglobin-A CD8 T cell epitopes

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Abstract

Mammaglobin-A (MAM-A) is a secretory protein that is overexpressed in 80 % of human breast cancers. Its near-universal expression in breast cancer as well as its exquisite tissue specificity makes it an attractive target for a breast cancer prevention vaccine, and we recently initiated a phase 1 clinical trial of a MAM-A DNA vaccine. Previously, we have identified multiple MAM-A CD8 T cell epitopes using a reverse immunology candidate epitope approach based on predicted binding, but to date no attempt has been made to identify epitopes using an unbiased approach. In this study, we used human T cells primed in vitro with autologous dendritic cells expressing MAM-A to systematically identify MAM-A CD8 T cell epitopes. Using this unbiased approach, we identified three novel HLA-A2-restricted MAM-A epitopes. CD8 T cells specific for these epitopes are able to recognize and lyse human breast cancer cells in a MAM-A-specific, HLA-A2-dependent fashion. HLA-A2+/MAM-A+ breast cancer patients have an increased prevalence of CD8 T cells specific for these novel MAM-A epitopes, and vaccination with a MAM-A DNA vaccine significantly increases the number of these CD8 T cells. The identification and translational validation of novel MAM-A epitopes has important implications for the ongoing clinical development of vaccine strategies targeting MAM-A. The novel MAM-A epitopes represent attractive targets for epitope-based vaccination strategies, and can also be used to monitor immune responses. Taken together these studies provide additional support for MAM-A as an important therapeutic target for the prevention and treatment of breast cancer.

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Acknowledgments

This work was supported by research Grants from the Swiss National Foundation (SNF) (PBBSP3-138709) (SM), DOD/CDMRP-BCRP W81XWH-06-1-0677 (WEG), and the Barnes Jewish Hospital Foundation (TM, SPG). We are also extremely grateful to George and Diana Holway, whose generous gift provided the resources necessary to help us realize the promise of new discovery.

Conflict of interest

The authors declare that they have no conflict of interest. All the experiments conducted in this study comply with the current laws of the United States of America. Peripheral blood from patients was obtained after informed consent in accordance with a protocol approved by the Institutional Review Board at Washington University School of Medicine (WUSM).

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Authors and Affiliations

  1. Department of Surgery, Washington University School of Medicine, St. Louis, MO, USA

    S. D. Soysal, S. Muenst, X. Zhang, T. Fleming, T. Mohanakumar, L. Li, J. Herndon, S. P. Goedegebuure & W. E. Gillanders

  2. Department of Surgery, University Hospital Basel, Basel, Switzerland

    S. D. Soysal & D. Oertli

  3. Institute of Pathology, University Hospital Basel, Schoenbeinstrasse 40, 4031, Basel, Switzerland

    S. Muenst

  4. Department of Biological Sciences and Border Biomedical Research Center, The University of Texas at El Paso, El Paso, TX, USA

    J. Kan-Mitchell & E. Huarte

  5. Department of Obstetrics & Gynecology, Washington University School of Medicine, St. Louis, MO, USA

    I. Wilkinson-Ryan

  6. The Alvin J. Siteman Cancer Center at Barnes-Jewish Hospital and Washington University School of Medicine, St. Louis, MO, USA

    T. Fleming, T. Mohanakumar, S. P. Goedegebuure & W. E. Gillanders

  7. Department of Biological Sciences, Tennessee State University, Nashville, TN, USA

    V. Tiriveedhi

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  1. S. D. Soysal
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  2. S. Muenst
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  3. J. Kan-Mitchell
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  4. E. Huarte
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  7. T. Fleming
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  8. V. Tiriveedhi
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  9. T. Mohanakumar
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  10. L. Li
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  11. J. Herndon
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  13. S. P. Goedegebuure
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  14. W. E. Gillanders
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Corresponding author

Correspondence to S. Muenst.

Additional information

Soysal SD and Muenst S contributed equally.

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10549_2014_3129_MOESM1_ESM.tif

Supplementary material 1 (TIFF 1910 kb). Supplementary Fig. 1. a, b PBMC from MAM-A+, HLA-A2+ breast cancer patients were stimulated with selected peptides and assessed for recognition of peptide-pulsed APC. IFNγ ELISPOT data from two patients are shown c Recognition of predicted minimal epitopes by peptide-specific T cells. Using the epitope prediction algorithm netMHC3.2, the best predicted minimal epitopes encoded by P2, P5, P12, and P13 were tested for recognition by peptide-specific T cells from two breast cancer patients in IFNγ ELISPOT assays

10549_2014_3129_MOESM2_ESM.tif

Supplementary material 2 (TIFF 1118 kb). Supplementary Fig. 2 a P2-specific T cells recognize AU565, and the HLA-A2+, MAM-A-negative MCF7 after pulsing with P2. Recognition of peptide-pulsed MCF7 was blocked by anti-HLA-A2 Ab (+1) b P12-specific T cells recognize AU565 and P12-pulsed MCF7 (+) but not MCF7 pulsed with P2 (+2)

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Soysal, S.D., Muenst, S., Kan-Mitchell, J. et al. Identification and translational validation of novel mammaglobin-A CD8 T cell epitopes. Breast Cancer Res Treat 147, 527–537 (2014). https://doi.org/10.1007/s10549-014-3129-x

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