Document Detail


On-chip activation and subsequent detection of individual antigen-specific T cells.
MedLine Citation:
PMID:  20000848     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Qing Song; Qing Han; Elizabeth M Bradshaw; Sally C Kent; Khadir Raddassi; Björn Nilsson; Gerald T Nepom; David A Hafler; J Christopher Love
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Analytical chemistry     Volume:  82     ISSN:  1520-6882     ISO Abbreviation:  Anal. Chem.     Publication Date:  2010 Jan 
Date Detail:
Created Date:  2010-01-14     Completed Date:  2010-03-22     Revised Date:  2014-09-24    
Medline Journal Info:
Nlm Unique ID:  0370536     Medline TA:  Anal Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  473-7     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Antibodies / immunology,  metabolism
CD4-Positive T-Lymphocytes / immunology*
Histocompatibility Antigens Class II / immunology,  metabolism
Humans
Interferon-gamma / metabolism
Interleukin-17 / metabolism
Lab-On-A-Chip Devices
Peptides / metabolism
Tissue Array Analysis / instrumentation,  methods*
Grant Support
ID/Acronym/Agency:
5U19AI050864-07/AI/NIAID NIH HHS; U19 AI050864/AI/NIAID NIH HHS; U19 AI050864-07/AI/NIAID NIH HHS
Chemical
Reg. No./Substance:
0/Antibodies; 0/Histocompatibility Antigens Class II; 0/Interleukin-17; 0/Peptides; 82115-62-6/Interferon-gamma
Comments/Corrections

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