On-chip activation and subsequent detection of individual antigen-specific T cells
(2010) In Analytical Chemistry 82(2). p.7-473- Abstract
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... (More)
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 (IFNgamma 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.
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- author
- Song, Qing ; Han, Qing ; Bradshaw, Elizabeth M ; Kent, Sally C ; Raddassi, Khadir ; Nilsson, Björn LU ; Nepom, Gerald T ; Hafler, David A and Love, J Christopher
- publishing date
- 2010-01-15
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Antibodies, CD4-Positive T-Lymphocytes, Histocompatibility Antigens Class II, Humans, Interferon-gamma, Interleukin-17, Lab-On-A-Chip Devices, Peptides, Tissue Array Analysis, Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't
- in
- Analytical Chemistry
- volume
- 82
- issue
- 2
- pages
- 5 pages
- publisher
- The American Chemical Society (ACS)
- external identifiers
-
- scopus:75649101297
- pmid:20000848
- ISSN
- 1520-6882
- DOI
- 10.1021/ac9024363
- language
- English
- LU publication?
- no
- id
- 47c74748-1942-496d-90f1-e40ac4e7d667
- date added to LUP
- 2016-10-27 13:54:35
- date last changed
- 2024-01-04 15:02:10
@article{47c74748-1942-496d-90f1-e40ac4e7d667, abstract = {{<p>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 (IFNgamma 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.</p>}}, author = {{Song, Qing and Han, Qing and Bradshaw, Elizabeth M and Kent, Sally C and Raddassi, Khadir and Nilsson, Björn and Nepom, Gerald T and Hafler, David A and Love, J Christopher}}, issn = {{1520-6882}}, keywords = {{Antibodies; CD4-Positive T-Lymphocytes; Histocompatibility Antigens Class II; Humans; Interferon-gamma; Interleukin-17; Lab-On-A-Chip Devices; Peptides; Tissue Array Analysis; Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't}}, language = {{eng}}, month = {{01}}, number = {{2}}, pages = {{7--473}}, publisher = {{The American Chemical Society (ACS)}}, series = {{Analytical Chemistry}}, title = {{On-chip activation and subsequent detection of individual antigen-specific T cells}}, url = {{http://dx.doi.org/10.1021/ac9024363}}, doi = {{10.1021/ac9024363}}, volume = {{82}}, year = {{2010}}, }