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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, Björn LU ; Nepom, Gerald T; Hafler, David A and Love, J Christopher (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
publishing date
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
external identifiers
  • scopus:75649101297
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
2018-05-29 09:37:42
@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},
  keyword      = {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},
  series       = {Analytical Chemistry},
  title        = {On-chip activation and subsequent detection of individual antigen-specific T cells},
  url          = {http://dx.doi.org/10.1021/ac9024363},
  volume       = {82},
  year         = {2010},
}