title: On-chip activation and subsequent detection of individual antigen-specific T cells.
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 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.affiliation: Department of Chemical Engineering, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: Qing
authors.initials: Q
authors.lastname: Song
authors.forename: Qing
authors.initials: Q
authors.lastname: Han
authors.forename: Elizabeth M
authors.initials: EM
authors.lastname: Bradshaw
authors.forename: Sally C
authors.initials: SC
authors.lastname: Kent
authors.forename: Khadir
authors.initials: K
authors.lastname: Raddassi
authors.forename: Björn
authors.initials: B
authors.lastname: Nilsson
authors.forename: Gerald T
authors.initials: GT
authors.lastname: Nepom
authors.forename: David A
authors.initials: DA
authors.lastname: Hafler
authors.forename: J Christopher
authors.initials: JC
authors.lastname: Love
mesh_terms: D000906:Antibodies / Q000276:immunology / Q000378:metabolism; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D000949:Histocompatibility Antigens Class II / Q000276:immunology / Q000378:metabolism; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism; D020381:Interleukin-17 / Q000378:metabolism; D056656:Lab-On-A-Chip Devices; D010455:Peptides / Q000378:metabolism; D046888:Tissue Array Analysis / Q000295:instrumentation / Q000379:methods*
chemical_list: D000906:Antibodies; D000949:Histocompatibility Antigens Class II; D020381:Interleukin-17; D010455:Peptides; D007371:Interferon-gamma