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.

Abstract

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 E7(11-20) 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 E7(11-20) peptide-pulsed dendritic cells. Cytolytic activity was determined by a standard 4-h (51)Cr-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 E7(11-20) 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.

• Authors: W M Schoell, R Mirhashemi, B Liu, M F Janicek, E R Podack, M A Penalver, H E Averette
• Journal: Gynecologic oncology (Gynecol Oncol) Vol. 74 Issue 3 Pg. 448-55 (Sep 1999) ISSN: 0090-8258 [Print] United States
• PMID: 10479508 (Publication Type: Journal Article)
• Copyright: Copyright 1999 Academic Press.

Chemical References

• Oncogene Proteins, Viral
• Papillomavirus E7 Proteins
• oncogene protein E7, Human papillomavirus type 16

Topics

• Dendritic Cells (immunology)
• Female
• Humans
• Immunotherapy (methods)
• Oncogene Proteins, Viral (immunology)
• Papillomaviridae (immunology)
• Papillomavirus E7 Proteins
• Species Specificity
• T-Lymphocytes, Cytotoxic
• Tumor Cells, Cultured
• Uterine Cervical Neoplasms (therapy)