High Avidity Melanoma-reactive Cytotoxic T Lymphocytes Are Efficiently Induced from Peripheral Blood Lymphocytes on Stimulation by Peptide-pulsed Melanoma Cells

Nadine Gervois (1, 2, 3) , Nathalie Labarrière (1, 2, 3) , Soizic Le Guiner (1, 2, 3) , Marie-Christine Pandolfino (2, 1, 3) , Jean-François Fonteneau (1, 2, 3) , Yannik Guilloux (1, 2, 3) , Elisabeth Diez (1, 2, 3) , Brigitte Dréno (3, 1, 2) , Francine Jotereau (1, 2, 3)

1 Interactions récepteurs ligands en immunocancérologie et immunopathologie
2 UTCG - Unité de Thérapie Cellulaire et Génique [CHU Nantes]
3 Service de dermatologie [Nantes]

To design an efficient procedure to expand high avidity melanoma reactive T cells and to perform immunotherapies, we compared conditions of peripheral blood lymphocyte (PBL) stimulation by Melan-A/MART-1 peptides. Avidity of induced CTLs was evaluated by measuring their lysis and cytokine secretion to peptide-pulsed transporter-associated protein-deficient cells and to melanoma cells. In side-by-side experiments, we show that melanoma cells, either allogeneic or autologous, induced the growth of high avidity Melan-A-reactive CTLs from all donors, whereas essentially low avid-ity T cells were induced by peptide-pulsed PBLs. We also show that at least two cytokines, interleukin-6 and interleu-kin-2, were required to promote the growth of high avidity CTLs. Once sorted by tetramer labeling or cloning, the specificity and reactivity to tumor cells of peptide-specific T cells induced by allogeneic melanoma cells were confirmed. We then describe a relatively simple and efficient procedure that allowed us to obtain systematically high amounts (in the range of billion) of high avidity Melan-A/ MART-1-specific T cells from the PBLs of HLA-A2 mela-noma patients and healthy donors in 3 months. Because this antigen is expressed by most melanoma tumors, this procedure should be useful for checking the efficiency of adoptive immunotherapy of melanoma tumors and using functionally well-defined Melan-A/MART-1-specific CTLs in a large group of patients.

Domaines: Immunologie; Immunothérapie; Cancer

Nadine Gervois, Nathalie Labarrière, Soizic Le Guiner, Marie-Christine Pandolfino, Jean-François Fonteneau, et al.. High Avidity Melanoma-reactive Cytotoxic T Lymphocytes Are Efficiently Induced from Peripheral Blood Lymphocytes on Stimulation by Peptide-pulsed Melanoma Cells. Clinical Cancer Research, 2000. ⟨inserm-02482162⟩