Identification of a major histocompatibility complex class I-restricted T-cell epitope in the tumour-associated antigen, 5T4.

2.50
Hdl Handle:
http://hdl.handle.net/10541/72715
Title:
Identification of a major histocompatibility complex class I-restricted T-cell epitope in the tumour-associated antigen, 5T4.
Authors:
Redchenko, Irina; Harrop, Richard; Ryan, Matthew G; Hawkins, Robert E; Carroll, Miles W
Abstract:
5T4 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.
Affiliation:
Oxford BioMedica (UK) Ltd, Medawar Centre, Oxford Science Park, Oxford, UK. i.redchenko@oxfordbiomedica.co.uk
Citation:
Identification of a major histocompatibility complex class I-restricted T-cell epitope in the tumour-associated antigen, 5T4. 2006, 118 (1):50-7 Immunology
Journal:
Immunology
Issue Date:
May-2006
URI:
http://hdl.handle.net/10541/72715
DOI:
10.1111/j.1365-2567.2006.02338.x
PubMed ID:
16630022
Type:
Article
Language:
en
ISSN:
0019-2805
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
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.en
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

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