Human T cell responses to HPV 16 E2 generated with monocyte‐derived dendritic cells

• Published in: International journal of cancer Vol. 94; no. 6; pp. 807 - 812
• Main Authors: Davidson, Emma J., Brown, Michael D., Burt, Deborah J., Parish, Joanna L., Gaston, Kevin, Kitchener, Henry C., Stacey, Simon N., Stern, Peter L.
• Format: Journal Article
• Language: English
• Published: New York John Wiley & Sons, Inc 15-12-2001; Wiley-Liss
• Subjects: Biological and medical sciences; cervical cancer; cervical intraepithelial neoplasia (CIN); cytotoxic T lymphocytes (CTL); Cytotoxicity, Immunologic; dendritic cells; Dendritic Cells - physiology; DNA-Binding Proteins; E2 protein; Female genital diseases; Gynecology. Andrology. Obstetrics; HLA-A2 Antigen - analysis; Host-tumor relations. Immunology. Biological markers; human papillomavirus (HPV); Humans; Medical sciences; Monocytes - physiology; Oncogene Proteins, Viral - immunology; primary immune responses; T-Lymphocytes, Cytotoxic - immunology; Tumors; vaccines; γ‐interferon ELISPOT; Host tumor relation; Carcinoma; Cellular immunity; Papovaviridae; Uterine cervix; Carcinogenesis; Papillomavirus; Human papillomavirus 16; Human papillomavirus; Antigen presenting cell; Malignant transformation; Established cell line; T-Lymphocyte; Female; Human; Dendritic cell; Immune response; Monocyte; Vaccine; Malignant tumor; In vitro; Female genital diseases; Infection; Virus; Viral disease; Intraepithelial neoplasia; Uterine cervix diseases
• ISSN: 0020-7136; 1097-0215
• DOI: 10.1002/ijc.1558

Persistent infection with human papillomavirus (HPV) type 16 has been implicated in the etiology of cervical cancer. The E2 protein is required early in viral infection and therefore may serve as a useful immune target for a vaccine aimed at prevention or therapy of premalignant lesions. Dendritic cells (DC) prepared from monocytes and pulsed with bacterially produced HPV 16 E2 C‐terminus protein were used to stimulate autologous T cells over several rounds of stimulation. T cells were tested for γ‐interferon release by ELISPOT and for cytotoxic activity by 51chromium release assays. To generate E2‐expressing target cells for cytotoxicity assays, we constructed a recombinant vaccinia virus encoding HPV 16 E2, which was used to infect autologous Epstein‐Barr virus‐transformed lymphoblastoid cell lines (LCL). The results show that DC pulsed with E2 C‐terminus protein induce γ‐interferon‐releasing T cells as demonstrated by ELISPOT. Furthermore, we demonstrate E2‐specific lysis of vaccinia‐E2 infected autologous LCL by CD8+ cytotoxic T lymphocytes (CTL). E2‐specific CTL did not lyse untreated autologous LCL or LCL infected with wild‐type vaccinia and showed low levels of cytotoxicity against natural killer cell‐sensitive K562 cells. In addition, T cells stimulated with DC in the absence of E2 failed to demonstrate lysis of vaccinia‐E2‐labeled targets. Phenotypically, CTL populations were CD3+/CD8+. These results will facilitate the study of naturally occurring T‐cell responses to HPV E2 in patients with cervical intraepithelial neoplasia and the development of immunotherapeutic strategies designed to treat this and other HPV‐associated diseases. © 2001 Wiley‐Liss, Inc.