Generation of Tumor-Specific Cytotoxic T Lymphocytes by Stimulation with HPV Type 16 E7 Peptide-Pulsed Dendritic Cells: An Approach to Immunotherapy of Cervical Cancer

• Published in: Gynecologic oncology Vol. 74; no. 3; pp. 448 - 455
• Main Authors: Schoell, Wolfgang M.J., Mirhashemi, Ramin, Liu, Bai, Janicek, Mike F., Podack, Eckhard R., Penalver, Manuel A., Averette, Hervy E.
• Format: Journal Article
• Language: English
• Published: San Diego, CA Elsevier Inc 01.09.1999; Elsevier
• Subjects: Antineoplastic agents; Biological and medical sciences; Dendritic Cells - immunology; Female; Humans; Immunotherapy; Immunotherapy - methods; Medical sciences; Oncogene Proteins, Viral - immunology; Papillomaviridae - immunology; Papillomavirus E7 Proteins; Pharmacology. Drug treatments; Species Specificity; T-Lymphocytes, Cytotoxic; Tumor Cells, Cultured; Uterine Cervical Neoplasms - therapy; Human; Dendritic cell; Effectory cell; Cell culture; Carcinoma; Peptides; Cytotoxicity; Papovaviridae; Malignant tumor; Uterine cervix; In vitro; Female genital diseases; Papillomavirus; Virus; Human papillomavirus 16; Human papillomavirus; Treatment; Immunotherapy; T-Lymphocyte; Female; Uterine cervix diseases
• ISSN: 0090-8258; 1095-6859
• DOI: 10.1006/gyno.1999.5504

Objective. The aim of this study was to generate HPV-16 E7 peptide-specific cytotoxic T lymphocytes (CTLs) in vitro for future adoptive immunotherapy of cervical cancer. Methods. Peripheral blood mononuclear cells (PBMC) were isolated from HLA-A2+ healthy donors. The PBMCs were incubated with HPV-16 E711–20 peptide and varying cytokines in the primary culture. Restimulation was performed weekly with peptide-pulsed, irradiated autologous PBMCs. Alternatively, the PBMCs were depleted of abundant CD4+ cells and stimulated with HPV-16 E711–20 peptide-pulsed dendritic cells. Cytolytic activity was determined by a standard 4-h 51Cr-release assay. Results. After 6 weeks in culture, we were able to establish peptide-specific CTL lines in one of seven donors by incubating PBMCs with HPV-16 E711–20 peptide. When we employed autologous peptide-pulsed dendritic cells to stimulate CD8+ cell-enriched PBMCs, we obtained CTL lines in four of seven donors. The primed CTLs were able to lyse the HLA-A2+ and HPV-16+ cervical cancer cell line Caski. SiHa, an HLA-A2-, but HPV 16+, cervical cancer cell line could be lysed only after transfection with HLA-A2. In addition, a high cytotoxicity (>80%) was obtained against peptide-pulsed, but not unpulsed, targets such as autologous Ebstein–Barr virus-immortalized B cells or allogeneic lipopolysaccaride-stimulated PBMCs. DCs were clearly the most potent of all tested antigen presenting cells to stimulate a CTL response in a proliferation assay. Conclusion. HPV-16 E7 peptide-specific CTLs could be generated in vitro. A practical protocol to expand the CTLs to a sufficient number for an application in a clinical trial is in progress.