An antigen-targeted approach to adoptive transfer therapy of cancer

• Published in: Cancer research (Chicago, Ill.) Vol. 59; no. 9; pp. 2167 - 2173
• Main Authors: VALMORI, D, PITTET, M. J, RIMOLDI, D, LIENARD, D, DUNBAR, R, CERUNDOLO, V, LEJEUNE, F, CEROTTINI, J.-C, ROMERO, P
• Format: Journal Article
• Language: English
• Published: Philadelphia, PA American Association for Cancer Research 01.05.1999
• Subjects: Antigens, Neoplasm - immunology; Antineoplastic agents; Biological and medical sciences; Biotinylation; Cell Line; Cell Separation; Cytotoxicity, Immunologic; Flow Cytometry; Fluorescent Antibody Technique, Indirect; HLA-A2 Antigen - immunology; Humans; Immunodominant Epitopes - immunology; Immunotherapy; Immunotherapy, Adoptive; Interleukin-2 - pharmacology; Lymphocyte Activation; Macromolecular Substances; MART-1 Antigen; Medical sciences; Melanoma - blood; Melanoma - immunology; Neoplasm Proteins - immunology; Peptide Fragments - immunology; Pharmacology. Drug treatments; Recombinant Fusion Proteins - pharmacology; T-Lymphocytes, Cytotoxic - cytology; T-Lymphocytes, Cytotoxic - immunology; T-Lymphocytes, Cytotoxic - transplantation; Tumor Cells, Cultured; Human; HLA-System; Tumor associated antigen; Antigenic determinant; T-Lymphocyte; Malignant melanoma; Cytotoxicity; Malignant tumor; In vitro; Adoptive transfer
• ISSN: 0008-5472; 1538-7445

Previous attempts to treat human malignancies by adoptive transfer of tumor-specific CTLs have been limited by the difficulty of isolating T cells of defined antigen specificity. The recent development of MHC class I/antigenic peptide tetrameric complexes that allow direct identification of antigen-specific T cells has opened new possibilities for the isolation and in vitro expansion of tumor-specific T cells. In the present study, we have derived polyclonal monospecific cell lines from circulating Melan-A-specific CTL precursors of HLA-A*0201+ melanoma patients by combining stimulation with recently identified peptide analogues of the immunodominant epitope from the melanoma-associated antigen Melan-A with staining with fluorescent HLA-A*0201/Melan-A peptide tetramers. In vitro expansion of antigen-specific CD8+ T cells was monitored by flow cytometry with the fluorescent tetramers and anti-CD8 monoclonal antibody. This analysis revealed that Melan-A 26-35 peptide analogues were much more efficient than the parental peptides in stimulating a rapid in vitro expansion of antigen-specific CD8+ T cells. These cells were then isolated by tetramer-guided cell sorting and subsequently expanded in vitro by mitogen stimulation. The resulting polyclonal but monospecific CTLs fully cross-recognized the parental peptides and were able to efficiently lyse Melan-A-expressing tumor cells. Altogether, these results pave the way to a molecularly defined approach to antigen-specific adoptive transfer therapy of cancer.