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dc.contributor.authorRedchenko, Irina
dc.contributor.authorHarrop, Richard
dc.contributor.authorRyan, Matthew G
dc.contributor.authorHawkins, Robert E
dc.contributor.authorCarroll, Miles W
dc.date.accessioned2009-07-07T10:40:42Z
dc.date.available2009-07-07T10:40:42Z
dc.date.issued2006-05
dc.identifier.citationIdentification of a major histocompatibility complex class I-restricted T-cell epitope in the tumour-associated antigen, 5T4. 2006, 118 (1):50-7 Immunologyen
dc.identifier.issn0019-2805
dc.identifier.pmid16630022
dc.identifier.doi10.1111/j.1365-2567.2006.02338.x
dc.identifier.urihttp://hdl.handle.net/10541/72715
dc.description.abstract5T4 is a surface glycoprotein expressed on placental trophoblasts and also on a wide range of human carcinomas. Its highly restricted expression on normal tissues and broad distribution on many carcinomas make 5T4 a promising target for cancer immunotherapy. In the current study, we set out to investigate whether a 5T4-specific cytotoxic T lymphocyte (CTL) repertoire exists in healthy individuals. CD4-depleted peripheral blood mononuclear cells (PBMCs) from blood donors were screened using an ex vivo interferon-gamma (IFN-gamma) enzyme-linked immunospot (ELISPOT) assay. A panel of overlapping peptides, spanning the full length of the 5T4 protein, was used as a source of antigen. In the process of screening, one out of 30 blood donors demonstrated a positive ex vivo IFN-gamma ELISPOT response to a single 5T4 peptide. A polyclonal T-cell line was derived from this donor by culturing PBMCs with autologous peptide-pulsed dendritic cells (DCs). The resulting polyclonal T-cell line and clones were tested in a 51Cr-release assay and by ELISPOT and were shown to be peptide specific. Furthermore, antigen-presenting cells (APCs), infected with a viral vector expressing 5T4, were able to stimulate IFN-gamma production by the peptide-specific T-cell clones. A minimal CD8 epitope, PLADLSPFA, has been identified and found to be restricted through human leucocyte antigen (HLA) Cw7. Subsequently, we have demonstrated that HLA-Cw7-positive colorectal cancer patients vaccinated with a recombinant vaccinia viral vector encoding 5T4 (TroVax) are capable of mounting a strong IFN-gamma ELISPOT response to this novel CTL epitope. These findings have potential application in cancer immunotherapy in terms of subunit vaccine design and the monitoring of immune responses induced in patients by 5T4-based therapies.
dc.language.isoenen
dc.subject.meshCancer Vaccines
dc.subject.meshCell Division
dc.subject.meshCells, Cultured
dc.subject.meshColorectal Neoplasms
dc.subject.meshEnzyme-Linked Immunosorbent Assay
dc.subject.meshEpitopes, T-Lymphocyte
dc.subject.meshGenes, MHC Class I
dc.subject.meshHistocompatibility Testing
dc.subject.meshHumans
dc.subject.meshInterferon-gamma
dc.subject.meshMembrane Glycoproteins
dc.subject.meshT-Lymphocytes, Cytotoxic
dc.titleIdentification of a major histocompatibility complex class I-restricted T-cell epitope in the tumour-associated antigen, 5T4.en
dc.typeArticleen
dc.contributor.departmentOxford BioMedica (UK) Ltd, Medawar Centre, Oxford Science Park, Oxford, UK. i.redchenko@oxfordbiomedica.co.uken
dc.identifier.journalImmunologyen
html.description.abstract5T4 is a surface glycoprotein expressed on placental trophoblasts and also on a wide range of human carcinomas. Its highly restricted expression on normal tissues and broad distribution on many carcinomas make 5T4 a promising target for cancer immunotherapy. In the current study, we set out to investigate whether a 5T4-specific cytotoxic T lymphocyte (CTL) repertoire exists in healthy individuals. CD4-depleted peripheral blood mononuclear cells (PBMCs) from blood donors were screened using an ex vivo interferon-gamma (IFN-gamma) enzyme-linked immunospot (ELISPOT) assay. A panel of overlapping peptides, spanning the full length of the 5T4 protein, was used as a source of antigen. In the process of screening, one out of 30 blood donors demonstrated a positive ex vivo IFN-gamma ELISPOT response to a single 5T4 peptide. A polyclonal T-cell line was derived from this donor by culturing PBMCs with autologous peptide-pulsed dendritic cells (DCs). The resulting polyclonal T-cell line and clones were tested in a 51Cr-release assay and by ELISPOT and were shown to be peptide specific. Furthermore, antigen-presenting cells (APCs), infected with a viral vector expressing 5T4, were able to stimulate IFN-gamma production by the peptide-specific T-cell clones. A minimal CD8 epitope, PLADLSPFA, has been identified and found to be restricted through human leucocyte antigen (HLA) Cw7. Subsequently, we have demonstrated that HLA-Cw7-positive colorectal cancer patients vaccinated with a recombinant vaccinia viral vector encoding 5T4 (TroVax) are capable of mounting a strong IFN-gamma ELISPOT response to this novel CTL epitope. These findings have potential application in cancer immunotherapy in terms of subunit vaccine design and the monitoring of immune responses induced in patients by 5T4-based therapies.


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