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Title: Combining MHC tetramer and intracellular cytokine staining for CD8(+) T cells to reveal antigenic epitopes naturally presented on tumor cells.
Authors: Dimopoulos, Nektaria;Jackson, Heather M;Ebert, Lisa;Guillaume, Philippe;Luescher, Immanuel F;Ritter, Gerd;Chen, Weisan
Affiliation: Ludwig Institute for Cancer Research, Melbourne Branch, Austin Health, Heidelberg, VIC 3084, Australia
Issue Date: 26-Oct-2008
Citation: Journal of Immunological Methods 2008; 340(1): 90-4
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.
Internal ID Number: 18957296
DOI: 10.1016/j.jim.2008.09.023
Type: Journal Article
Subjects: Antigens, Neoplasm.analysis.immunology
CD8-Positive T-Lymphocytes.immunology
Cell Line, Tumor
Epitopes, T-Lymphocyte.analysis.immunology
Flow Cytometry
Histocompatibility Antigens.immunology
Appears in Collections:Journal articles

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