Generation of Anti-p53 Cytotoxic T Lymphocytes from Human Peripheral Blood Using Autologous Dendritic Cells

• Published in: Clinical cancer research 1999-06, Vol.5 (6), p.1281-1288
• Main Authors: CHIKAMATSU, K, NAKANO, K, STORKUS, W. J, APPELLA, E, LOTZE, M. T, WHITESIDE, T. L, DELEO, A. B
• Format: Article
• Language: English
• Subjects: Antigens, CD - metabolism; Antineoplastic agents; Biological and medical sciences; CD8-Positive T-Lymphocytes - immunology; Cell Separation; Cells, Cultured; Clone Cells - immunology; Cytotoxicity Tests, Immunologic; Dendritic Cells - immunology; Epitopes - immunology; HLA-A2 Antigen - immunology; Humans; Immunotherapy; Interleukins - metabolism; Lymphocyte Subsets - immunology; Medical sciences; Peptide Fragments - immunology; Pharmacology. Drug treatments; T-Lymphocytes, Cytotoxic - immunology; T-Lymphocytes, Cytotoxic - metabolism; Tumor Cells, Cultured; Tumor Suppressor Protein p53 - genetics; Tumor Suppressor Protein p53 - immunology
• ISSN: 1078-0432; 1557-3265

CTLs recognizing the HLA-A2.1-restricted, wild-type sequence p53 epitopes p53 149–157 and p53 264–272 were generated from CD8-enriched populations of nonadherent peripheral blood lymphocytes (PBLs) obtained from healthy donors. The PBLs were restimulated in vitro with peptide-pulsed granulocyte macrophage colony-stimulating factor- and interleukin (IL)-4-induced autologous dendritic cells in the presence of IL-6 and IL-12 and subsequently cultivated with IL-1α, IL-2, IL-4, IL-6, and IL-7. Bulk anti-p53 264–272 CTL populations were generated from PBLs obtained from two of five donors. Both CTL populations were cytotoxic against peptide-pulsed HLA-A2 + target cells, but not against untreated target cells. A CD8 + anti-p53 CTL clone designated p264#2 was isolated from one of the bulk populations. It was found to have an intermediate affinity of approximately 10 −9 m for the epitope and to mediate cytotoxicity against several human tumor cell lines, including the squamous cell carcinoma of the head and neck cell line SCC-9, which is known to present the wild-type sequence p53 264–272 epitope. In addition, CTLs reactive against p53 149–157 -pulsed targets as well as a HLA-A2 + tumor cell line were cloned from a bulk population of antitumor CTLs obtained from one of the five normal PBLs restimulated with this epitope. The results indicate that CTLs recognizing wild-type sequence epitopes can be generated from precursors present in PBLs obtained from some normal individuals using autologous dendritic cells as antigen-presenting cells and suggest that vaccine strategies targeting these epitopes can lead to antitumor CTL generation, thereby emphasizing the therapeutic potential of p53-based cancer vaccines.