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Pascal Chaux^*, Valérie Vantomme^*, Vincent Stroobant^*, Kris Thielemans, Jurgen Corthals, Rosalie Luiten^*, Alexander M.M. Eggermont, Thierry Boon^*, and Pierre van der Bruggen^*

From the ^* Ludwig Institute for Cancer Research, Université Catholique de Louvain 74.59, B-1200 Brussels, Belgium; the Laboratory of Physiology, Medical School, Free University of Brussels, B-1070 Brussels, Belgium; and Erasmus University Rotterdam, Department of Surgical Oncology, University Hospital Rotterdam, Daniel den Hoed Cancer Center, 3075 EA Rotterdam, The Netherlands

MAGE-type genes are expressed by many tumors of different histological types and not by normal cells, except for male germline cells, which do not express major histocompatibility complex (MHC) molecules. Therefore, the antigens encoded by MAGE-type genes are strictly tumor specific and common to many tumors. We describe here the identification of the first MAGE-encoded epitopes presented by histocompatibility leukocyte antigen (HLA) class II molecules to CD4⁺ T lymphocytes. Monocyte-derived dendritic cells were loaded with a MAGE-3 recombinant protein and used to stimulate autologous CD4⁺ T cells. We isolated CD4⁺ T cell clones that recognized two different MAGE-3 epitopes, MAGE-3_114–127 and MAGE-3_121–134, both presented by the HLA-DR13 molecule, which is expressed in 20% of Caucasians. The second epitope is also encoded by MAGE-1, -2, and -6. Our procedure should be applicable to other proteins for the identification of new tumor-specific antigens presented by HLA class II molecules. The knowledge of such antigens will be useful for evaluation of the immune response of cancer patients immunized with proteins or with recombinant viruses carrying entire genes coding for tumor antigens. The use of antigenic peptides presented by class II in addition to peptides presented by class I may also improve the efficacy of therapeutic antitumor vaccination.

Key Words: human • invariant chain • peptide • tumor • histocompatibility leukocyte antigen class II

P. Chaux and R. Luiten were supported by a postdoctoral fellowship from the Training and Mobility of Researchers Program of the European Commission. V. Vantomme was partially supported by the Fonds National de la Recherche Scientifique (TELEVIE grants), Brussels, Belgium. This work was partially supported by the Belgian Programme on Interuniversity Poles of Attraction initiated by the Belgian State, Prime Minister's Office, Science Policy Programming, and by grants from the Association contre le Cancer, Brussels, Belgium, from the BIOMED 2 programme of the European Community, from the Fonds J. Maisin, Belgium, and from the Caisse Générale d'Epargne et de Retraite (CGER)-Assurances and VIVA, Brussels, Belgium.

Abbreviations used: aa, amino acid(s); Ii, invariant chain(s).

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