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Detection and enrichment of antigen-specific CD4+ and CD8+ T cells based on cytokine secretion

The Cytokine Secretion Assay is an innovative method for analysing and enriching live cytokine-secreting cells. In this assay, a cytokine affinity matrix is built on the cell plasma membrane, which traps cytokines produced by the cell in response to specific stimuli. The specifically bound cytokine...

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Published in:	Methods (San Diego, Calif.) Calif.), 2003-10, Vol.31 (2), p.150-159
Main Author:	Campbell, John D.M
Format:	Article
Language:	English
Subjects:	CD4-Positive T-Lymphocytes - cytology CD4-Positive T-Lymphocytes - immunology CD4-Positive T-Lymphocytes - secretion CD8-Positive T-Lymphocytes - cytology CD8-Positive T-Lymphocytes - immunology CD8-Positive T-Lymphocytes - secretion Cytokines - secretion Flow Cytometry Humans Immunomagnetic Separation - methods Interferon-gamma - secretion Receptors, Antigen, T-Cell - immunology Staining and Labeling - methods Viral Matrix Proteins - immunology
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container_title	Methods (San Diego, Calif.)
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creator	Campbell, John D.M
description	The Cytokine Secretion Assay is an innovative method for analysing and enriching live cytokine-secreting cells. In this assay, a cytokine affinity matrix is built on the cell plasma membrane, which traps cytokines produced by the cell in response to specific stimuli. The specifically bound cytokine is then detected, and the cells optionally enriched, using fluorochrome-conjugated cytokine-specific antibodies and magnetic microbeads. This method allows extremely detailed phenotyping of live cells and the detection of cytokine responses at very low frequencies. Here, the latest cell staining and separation procedures are reviewed, with particular reference to the best application of the technology and troubleshooting in a variety of different situations.
doi_str_mv	10.1016/S1046-2023(03)00125-7
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subjects	CD4-Positive T-Lymphocytes - cytology CD4-Positive T-Lymphocytes - immunology CD4-Positive T-Lymphocytes - secretion CD8-Positive T-Lymphocytes - cytology CD8-Positive T-Lymphocytes - immunology CD8-Positive T-Lymphocytes - secretion Cytokines - secretion Flow Cytometry Humans Immunomagnetic Separation - methods Interferon-gamma - secretion Receptors, Antigen, T-Cell - immunology Staining and Labeling - methods Viral Matrix Proteins - immunology
title	Detection and enrichment of antigen-specific CD4+ and CD8+ T cells based on cytokine secretion
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