On-Chip Activation and Subsequent Detection of Individual Antigen-Specific T Cells
The frequencies of antigen-specific CD4+ T cells in samples of human tissue have been difficult to determine accurately ex vivo, particularly for autoimmune diseases such as multiple sclerosis or type 1 diabetes. Conventional approaches involve the expansion of primary T cells in vitro to increase t...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | en_US |
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American Chemical Society
2013
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| Online Access: | http://hdl.handle.net/1721.1/79369 https://orcid.org/0000-0002-7989-2376 |
| _version_ | 1826208751874473984 |
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| author | Song, Qing Han, Qing Bradshaw, Elizabeth M. Kent, Sally C. Raddassi, Khadir Nilsson, Bjorn Nepom, Gerald T. Hafler, David A. Love, J. Christopher |
| author2 | Massachusetts Institute of Technology. Department of Chemical Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Chemical Engineering Song, Qing Han, Qing Bradshaw, Elizabeth M. Kent, Sally C. Raddassi, Khadir Nilsson, Bjorn Nepom, Gerald T. Hafler, David A. Love, J. Christopher |
| author_sort | Song, Qing |
| collection | MIT |
| description | The frequencies of antigen-specific CD4+ T cells in samples of human tissue have been difficult to determine accurately ex vivo, particularly for autoimmune diseases such as multiple sclerosis or type 1 diabetes. Conventional approaches involve the expansion of primary T cells in vitro to increase the numbers of cells, and a subsequent assessment of the frequencies of antigen-specific T cells in the expanded population by limiting dilution or by using fluorescently labeled tetramers of peptide-loaded major histocompatibility complex (MHC) receptors. Here we describe an alternative approach that uses arrays of subnanoliter wells coated with recombinant peptide-loaded MHC class II monomers to isolate and stimulate individual CD4+ T cells in an antigen-specific manner. In these experiments, activation was monitored using microengraving to capture two cytokines (IFNγ and IL-17) released from single cells. This new method should enable direct enumeration of antigen-specific CD4+ T cells ex vivo from clinical samples. |
| first_indexed | 2024-09-23T14:11:03Z |
| format | Article |
| id | mit-1721.1/79369 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T14:11:03Z |
| publishDate | 2013 |
| publisher | American Chemical Society |
| record_format | dspace |
| spelling | mit-1721.1/793692022-10-01T19:41:09Z On-Chip Activation and Subsequent Detection of Individual Antigen-Specific T Cells Song, Qing Han, Qing Bradshaw, Elizabeth M. Kent, Sally C. Raddassi, Khadir Nilsson, Bjorn Nepom, Gerald T. Hafler, David A. Love, J. Christopher Massachusetts Institute of Technology. Department of Chemical Engineering Massachusetts Institute of Technology. Department of Mechanical Engineering Song, Qing Han, Qing Love, Christopher J. The frequencies of antigen-specific CD4+ T cells in samples of human tissue have been difficult to determine accurately ex vivo, particularly for autoimmune diseases such as multiple sclerosis or type 1 diabetes. Conventional approaches involve the expansion of primary T cells in vitro to increase the numbers of cells, and a subsequent assessment of the frequencies of antigen-specific T cells in the expanded population by limiting dilution or by using fluorescently labeled tetramers of peptide-loaded major histocompatibility complex (MHC) receptors. Here we describe an alternative approach that uses arrays of subnanoliter wells coated with recombinant peptide-loaded MHC class II monomers to isolate and stimulate individual CD4+ T cells in an antigen-specific manner. In these experiments, activation was monitored using microengraving to capture two cytokines (IFNγ and IL-17) released from single cells. This new method should enable direct enumeration of antigen-specific CD4+ T cells ex vivo from clinical samples. National Institute of Allergy and Infectious Diseases (U.S.) (Award Number 5U19AI050864-07) Juvenile Diabetes Research Foundation International Massachusetts Institute of Technology (Texaco- Mangelsdorf Career Development Professor) 2013-06-26T15:09:09Z 2013-06-26T15:09:09Z 2009-12 2009-10 Article http://purl.org/eprint/type/JournalArticle 0003-2700 1520-6882 http://hdl.handle.net/1721.1/79369 Song, Qing, Qing Han, Elizabeth M. Bradshaw, Sally C. Kent, Khadir Raddassi, Björn Nilsson, Gerald T. Nepom, David A. Hafler, and J. Christopher Love. On-Chip Activation and Subsequent Detection of Individual Antigen-Specific T Cells. Analytical Chemistry 82, no. 2 (January 15, 2010): 473-477. https://orcid.org/0000-0002-7989-2376 en_US http://dx.doi.org/10.1021/ac9024363 Analytical Chemistry Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf American Chemical Society PMC |
| spellingShingle | Song, Qing Han, Qing Bradshaw, Elizabeth M. Kent, Sally C. Raddassi, Khadir Nilsson, Bjorn Nepom, Gerald T. Hafler, David A. Love, J. Christopher On-Chip Activation and Subsequent Detection of Individual Antigen-Specific T Cells |
| title | On-Chip Activation and Subsequent Detection of Individual Antigen-Specific T Cells |
| title_full | On-Chip Activation and Subsequent Detection of Individual Antigen-Specific T Cells |
| title_fullStr | On-Chip Activation and Subsequent Detection of Individual Antigen-Specific T Cells |
| title_full_unstemmed | On-Chip Activation and Subsequent Detection of Individual Antigen-Specific T Cells |
| title_short | On-Chip Activation and Subsequent Detection of Individual Antigen-Specific T Cells |
| title_sort | on chip activation and subsequent detection of individual antigen specific t cells |
| url | http://hdl.handle.net/1721.1/79369 https://orcid.org/0000-0002-7989-2376 |
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