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Japanese Journal of Clinical Oncology 30:117-121 (2000)
© 2000 Foundation for Promotion of Cancer Research

Analysis of Individual Specific Cytotoxic T Lymphocytes for Two MAGE-3-derived Epitopes Presented by HLA-A24

Fumihiro Katsura1, Masao Eura1, Kazuaki Chikamatsu1, Masatake Oiso1, Eiji Yumoto1 and Takeru Ishikawa2,+

1Department of Otorhinolaryngology, Kumamoto University School of Medicine, Kumamoto and 2Allergy and Immunology Center of Kyushu, Kumamoto, Japan

Background: The human MAGE-3 gene encodes tumor-specific antigens that are recognized by cytotoxic T lymphocytes (CTLs) and expressed in a high percentage of various malignant tumors. Of the five MAGE-3-derived CTL epitopes identified to date, two nonapeptides (TFPDLESEF and IMPKAGLLI, designated MAGE-3.A24a and MAGE-3.A24b, respectively) can be expressed on the tumor surface by binding to the HLA-A24 molecule, which is the most frequent HLA class I molecule in Asian populations. To compare the immunogenecities of the two peptides, individual specific CTL lines were generated for each peptide (MAGE-3.A24a and MAGE-3.A24b).

Methods: Peripheral blood mononuclear cells (PBMCs) from four HLA-A24+ healthy donors were stimulated in vitro with autologous dendritic cells pulsed with MAGE-3.A24a, MAGE-3.A24b or both and were subsequently cultivated with a cytokine combination including interleukin-2.

Results: We succeeded in generating peptide-specific CTL lines in two of the four donors. The two CTL lines showed similar cytolytic levels against three MAGE-3+/HLA-A24+ cancer cell lines and also target cells pulsed with the corresponding peptide. Cytolytic activities were blocked by either anti-CD8 or anti-HLA-A24 monoclonal antibodies.

Conclusions: The results suggest that MAGE-3.A24a and MAGE-3.A24b peptides have equal potential in inducing MAGE-3-specific and HLA-A24-restricted CTLs.

+ For reprints and all correspondence: Masao Eura, Department of Otorhinolaryngology, Kumamoto University School of Medicine, 1-1-1 Honjo, Kumamoto 860-8556, JapanAbbreviations: CTL, cytotoxic T lymphocyte; PBMC, peripheral blood mononuclear cell; SCCHN, squamous cell carcinoma of the head and neck; MHC, major histocompatibility complex; MAb, monoclonal antibody; PBS, phosphate-buffered saline; DC, dendritic cell; APC, antigen presenting cell


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