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

2.50
Hdl Handle:
http://hdl.handle.net/10541/85604
Title:
Human T cell responses to HPV 16 E2 generated with monocyte-derived dendritic cells.
Authors:
Davidson, Emma J; Brown, Michael D; Burt, Deborah J; Parish, Joanna L; Gaston, Kevin; Kitchener, Henry C; Stacey, Simon N; Stern, Peter L
Abstract:
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 gamma-interferon release by ELISPOT and for cytotoxic activity by (51)chromium 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 gamma-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.
Affiliation:
CRC Immunology Group, Paterson Institute for Cancer Research, Christie Hospital NHS Trust, Manchester, United Kingdom.
Citation:
Human T cell responses to HPV 16 E2 generated with monocyte-derived dendritic cells. 2001, 94 (6):807-12 Int. J. Cancer
Journal:
International Journal of Cancer.
Issue Date:
15-Dec-2001
URI:
http://hdl.handle.net/10541/85604
PubMed ID:
11745482
Type:
Article
Language:
en
ISSN:
0020-7136
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorDavidson, Emma Jen
dc.contributor.authorBrown, Michael Den
dc.contributor.authorBurt, Deborah Jen
dc.contributor.authorParish, Joanna Len
dc.contributor.authorGaston, Kevinen
dc.contributor.authorKitchener, Henry Cen
dc.contributor.authorStacey, Simon Nen
dc.contributor.authorStern, Peter Len
dc.date.accessioned2009-11-06T16:32:01Z-
dc.date.available2009-11-06T16:32:01Z-
dc.date.issued2001-12-15-
dc.identifier.citationHuman T cell responses to HPV 16 E2 generated with monocyte-derived dendritic cells. 2001, 94 (6):807-12 Int. J. Canceren
dc.identifier.issn0020-7136-
dc.identifier.pmid11745482-
dc.identifier.urihttp://hdl.handle.net/10541/85604-
dc.description.abstractPersistent 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 gamma-interferon release by ELISPOT and for cytotoxic activity by (51)chromium 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 gamma-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.en
dc.language.isoenen
dc.subject.meshCytotoxicity, Immunologic-
dc.subject.meshDNA-Binding Proteins-
dc.subject.meshDendritic Cells-
dc.subject.meshHLA-A2 Antigen-
dc.subject.meshHumans-
dc.subject.meshMonocytes-
dc.subject.meshOncogene Proteins, Viral-
dc.subject.meshT-Lymphocytes, Cytotoxic-
dc.titleHuman T cell responses to HPV 16 E2 generated with monocyte-derived dendritic cells.en
dc.typeArticleen
dc.contributor.departmentCRC Immunology Group, Paterson Institute for Cancer Research, Christie Hospital NHS Trust, Manchester, United Kingdom.en
dc.identifier.journalInternational Journal of Cancer.en

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