Combining MHC tetramer and intracellular cytokine staining for CD8(+) T cells to reveal antigenic epitopes naturally presented on tumor cells.

Details

Serval ID
serval:BIB_63FA473A8669
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Combining MHC tetramer and intracellular cytokine staining for CD8(+) T cells to reveal antigenic epitopes naturally presented on tumor cells.
Journal
Journal of immunological methods
Author(s)
Dimopoulos N., Jackson H.M., Ebert L., Guillaume P., Luescher I.F., Ritter G., Chen W.
ISSN
0022-1759
Publication state
Published
Issued date
2009
Peer-reviewed
Oui
Volume
340
Number
1
Pages
90-94
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't Publication Status: ppublish
Abstract
As more tumor antigens are discovered and as computer-guided T cell epitope prediction programs become more sophisticated, many potential T cell epitopes are synthesized and demonstrated to be antigenic in vitro. However, it is estimated that about 50% of such tumor antigen-specific T cells have not been demonstrated to recognize the naturally presented epitopes due to either technical difficulties, such as T cell cloning which is still challenging for many laboratories; or the predicted T cell epitopes are not generated or not generated in sufficient amounts by the antigen processing machinery. However, to potentially identify clinically relevant vaccine candidate epitopes, it is essential to demonstrate natural antigen presentation. Here we combine the advantages of MHC tetramer and intracellular cytokine staining to sensitively detect natural antigen presentation by tumor cells for epitopes of interest. The novel method does not require T cell cloning or long-term T cell culture. Because the antigen-specific T cells are positively identified, this method is much less influenced by IFNgamma producing cells with unknown specificities and should be widely applicable.
Keywords
Antigens, Neoplasm/analysis, Antigens, Neoplasm/immunology, CD8-Positive T-Lymphocytes/immunology, Cell Line, Tumor, Cytokines/analysis, Cytokines/immunology, Epitopes, T-Lymphocyte/analysis, Epitopes, T-Lymphocyte/immunology, Flow Cytometry, Histocompatibility Antigens/immunology, Humans, Melanoma/immunology
Pubmed
Web of science
Create date
15/01/2010 15:14
Last modification date
20/08/2019 14:20
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