Identification of HLA-A24-restricted CD8+ cytotoxic T-cell epitopes derived from mammaglobin-A, a human breast cancer–associated antigen

• Published in: Human immunology 2012-01, Vol.73 (1), p.11-16
• Main Authors: Tiriveedhi, Venkataswarup, Sarma, Nayan J, Subramanian, Vijay, Fleming, Timothy P, Gillanders, William E, Mohanakumar, Thallachallour
• Format: Article
• Language: English
• Subjects: Allergy and Immunology; Amino Acid Sequence; Binding, Competitive - immunology; Breast cancer; Breast Neoplasms - genetics; Breast Neoplasms - immunology; Breast Neoplasms - pathology; Cancer Vaccines - genetics; Cancer Vaccines - immunology; Cancer Vaccines - metabolism; CD8 Antigens - immunology; CD8 Antigens - metabolism; CD8 T cell; Cell Line, Tumor; Cells, Cultured; Cytotoxicity, Immunologic - immunology; Epitope Mapping; Epitopes, T-Lymphocyte - genetics; Epitopes, T-Lymphocyte - immunology; Epitopes, T-Lymphocyte - metabolism; Female; HLA-A24; HLA-A24 Antigen - genetics; HLA-A24 Antigen - immunology; HLA-A24 Antigen - metabolism; Humans; Mammaglobin A - genetics; Mammaglobin A - immunology; Mammaglobin-A; Protein Binding - immunology; T-Lymphocytes, Cytotoxic - immunology; T-Lymphocytes, Cytotoxic - metabolism; Transfection; Vaccine
• ISSN: 0198-8859; 1879-1166
• DOI: 10.1016/j.humimm.2011.10.017

Abstract Human breast cancer–associated antigen, mammaglobin-A (Mam-A), potentially offers a novel therapeutic target as a breast cancer vaccine. In this study, we define the CD8+ cytotoxic T lymphocyte (CTL) response to Mam-A-derived candidate epitopes presented in the context of HLA-A24 (A*2402). HLA-A24 has a frequency of 72% in Japanese, 27% in Asian Indian, and 18% in Caucasian populations. Using a human leukocyte antigen (HLA)-binding prediction algorithm we identified 7 HLA-A24-restricted Mam-A-derived candidate epitopes (MAA24.1–7). Membrane stabilization studies with TAP-deficient T2 cells transfected with HLA-A2402 (T2.A24) indicated that MAA24.2 (CYAGSGCPL) and MAA24.4 (ETLSNVEVF) have the highest HLA-A24 binding affinity. Further, 2 CD8+ CTL cell lines generated in vitro against T2.A24 cells individually loaded with Mam-A-derived candidate epitopes demonstrated significant cytotoxic activity against MAA24.2 and MAA24.4. In addition, the same CD8+ CTL lines lysed the HLA-A24+ /Mam-A+ stable transfected human breast cancer cell lines AU565 and MDA-MB-361. However, these CTLs had no cytotoxicity against HLA-A24- /Mam-A+ and HLA-A24+ /Mam-A- breast cancer cell lines. In summary, our results define HLA-A24-restricted, Mam-A-derived, CD8+ CTL epitopes that can potentially be employed for Mam-A-based breast cancer vaccine therapy to breast cancer patients with HLA-A24 phenotype.