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Examining the presentation of tumor-associated antigens on peptide-pulsed T2 cells

Peptide-pulsed T2 cells are routinely used to study T-cell activation by MHC-restricted peptides derived from tumor-associated antigens (TAAs). Nevertheless, the capacity of T2 cells to present antigenic epitopes remains to be precisely quantified, primarily due to the detection limits imposed by av...

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Autores principales: Bossi, Giovanna, Gerry, Andrew B, Paston, Samantha J, Sutton, Deborah H, Hassan, Namir J, Jakobsen, Bent K
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Taylor & Francis 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3894244/
https://www.ncbi.nlm.nih.gov/pubmed/24482751
http://dx.doi.org/10.4161/onci.26840
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author Bossi, Giovanna
Gerry, Andrew B
Paston, Samantha J
Sutton, Deborah H
Hassan, Namir J
Jakobsen, Bent K
author_facet Bossi, Giovanna
Gerry, Andrew B
Paston, Samantha J
Sutton, Deborah H
Hassan, Namir J
Jakobsen, Bent K
author_sort Bossi, Giovanna
collection PubMed
description Peptide-pulsed T2 cells are routinely used to study T-cell activation by MHC-restricted peptides derived from tumor-associated antigens (TAAs). Nevertheless, the capacity of T2 cells to present antigenic epitopes remains to be precisely quantified, primarily due to the detection limits imposed by available methods. Since naturally-processed TAA-derived epitopes have been shown to be displayed at levels as low as 10–150 copies per cell, highly sensitive detection and quantification techniques are essential to assess the natural degree of T-cell sensitivity. Here, we report the use of soluble, high-affinity T-cell receptors (TCRs) coupled with single-molecule fluorescence microscopy to quantify three reported TAA-derived epitopes on peptide-pulsed T2 cells, dissecting the relationship between concentration of exogenous peptide, number of epitopes presented, and activation of epitope-specific T cells. Our findings indicate that peptide concentrations in the low nanomolar range are required for T2 cells to present TAAs in extents that are comparable to those of malignant cells.
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spelling pubmed-38942442014-01-30 Examining the presentation of tumor-associated antigens on peptide-pulsed T2 cells Bossi, Giovanna Gerry, Andrew B Paston, Samantha J Sutton, Deborah H Hassan, Namir J Jakobsen, Bent K Oncoimmunology Brief Report Peptide-pulsed T2 cells are routinely used to study T-cell activation by MHC-restricted peptides derived from tumor-associated antigens (TAAs). Nevertheless, the capacity of T2 cells to present antigenic epitopes remains to be precisely quantified, primarily due to the detection limits imposed by available methods. Since naturally-processed TAA-derived epitopes have been shown to be displayed at levels as low as 10–150 copies per cell, highly sensitive detection and quantification techniques are essential to assess the natural degree of T-cell sensitivity. Here, we report the use of soluble, high-affinity T-cell receptors (TCRs) coupled with single-molecule fluorescence microscopy to quantify three reported TAA-derived epitopes on peptide-pulsed T2 cells, dissecting the relationship between concentration of exogenous peptide, number of epitopes presented, and activation of epitope-specific T cells. Our findings indicate that peptide concentrations in the low nanomolar range are required for T2 cells to present TAAs in extents that are comparable to those of malignant cells. Taylor & Francis 2013-10-21 /pmc/articles/PMC3894244/ /pubmed/24482751 http://dx.doi.org/10.4161/onci.26840 Text en Copyright © 2013 Landes Bioscience http://creativecommons.org/licenses/by-nc/3.0/ This is an open-access article licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported License. The article may be redistributed, reproduced, and reused for non-commercial purposes, provided the original source is properly cited.
spellingShingle Brief Report
Bossi, Giovanna
Gerry, Andrew B
Paston, Samantha J
Sutton, Deborah H
Hassan, Namir J
Jakobsen, Bent K
Examining the presentation of tumor-associated antigens on peptide-pulsed T2 cells
title Examining the presentation of tumor-associated antigens on peptide-pulsed T2 cells
title_full Examining the presentation of tumor-associated antigens on peptide-pulsed T2 cells
title_fullStr Examining the presentation of tumor-associated antigens on peptide-pulsed T2 cells
title_full_unstemmed Examining the presentation of tumor-associated antigens on peptide-pulsed T2 cells
title_short Examining the presentation of tumor-associated antigens on peptide-pulsed T2 cells
title_sort examining the presentation of tumor-associated antigens on peptide-pulsed t2 cells
topic Brief Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3894244/
https://www.ncbi.nlm.nih.gov/pubmed/24482751
http://dx.doi.org/10.4161/onci.26840
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