| ELISPOT and functional T cell analyses using HLA mono-specific target cells. | |
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MedLine Citation:
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PMID: 19729014 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Simple T cell assays specific for any chosen HLA class I or class II/peptide combination, are of enormous value in cancer immunotherapy, clinical trials, vaccine and infectious disease research. The reliable measurement of T cell activity can be difficult due to the presence of other alleles on target cells, particularly for the non-HLA-A2 alleles, and the varying baseline characteristics of the different APCs employed. In the absence of pulsing with HLA-A2 restricted peptides, T2 cells are functionally HLA class I and II negative. By coating these cells with recombinant HLA peptide complexes, HLA mono-specific cells are produced that present only a defined single epitope, and generate minimal background immune activation. In ELISPOT, intracellular cytokine staining (ICS) and killing assays using T cells specific for HLA-A2/peptide complexes, the HLA mono-specific cells gave comparable results, to those using standard peptide pulsed HLA-A2 positive T2 cells without significant background. Successful T cell assays for non-HLA-A2 T cells were also performed, with PBMCs recognizing HLA-A24 and HLA-DR15/peptide complexes. The data, obtained with ELISPOT, ICS and FACS-based killing assays, all demonstrate high specificity of T cell activity and low levels of background activity. HLA mono-specific cells are simple to prepare, and can be used with any stable recombinant HLA allele/peptide combination; providing a useful system for improved T cell functional analyses across all HLA allotypes. This represents a significant advance in the generation of reliable functional T cell data. |
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Authors:
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Claire Horlock; Bryony Stott; Julian Dyson; Graham Ogg; Tess McPherson; Louise Jones; Andrew K Sewell; Linda Wooldridge; David K Cole; Justin Stebbing; Philip Savage |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2009-09-01 |
Journal Detail:
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Title: Journal of immunological methods Volume: 350 ISSN: 1872-7905 ISO Abbreviation: J. Immunol. Methods Publication Date: 2009 Oct |
Date Detail:
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Created Date: 2009-10-14 Completed Date: 2009-11-09 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 1305440 Medline TA: J Immunol Methods Country: Netherlands |
Other Details:
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Languages: eng Pagination: 150-60 Citation Subset: IM |
Affiliation:
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Department of Immunology, Imperial College, London W12 0NN, United Kingdom. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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HLA-A Antigens HLA-DR Antigens / immunology* Humans Immunoenzyme Techniques / methods Peptides / immunology* T-Lymphocytes / immunology* |
| Chemical | |
Reg. No./Substance:
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0/HLA-A Antigens; 0/HLA-A24; 0/HLA-DR Antigens; 0/HLA-DR15; 0/Peptides |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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