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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; Nepom, Gerald T; Hafler, David A; Love, J Christopher.
Afiliação
  • Song Q; Department of Chemical Engineering, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
Anal Chem ; 82(2): 473-7, 2010 Jan 15.
Article em En | MEDLINE | ID: mdl-20000848
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.
Assuntos

Texto completo: 1 Coleções: 01-internacional Temas: Geral Base de dados: MEDLINE Assunto principal: Linfócitos T CD4-Positivos / Análise Serial de Tecidos Tipo de estudo: Diagnostic_studies Limite: Humans Idioma: En Revista: Anal Chem Ano de publicação: 2010 Tipo de documento: Article País de afiliação: Estados Unidos

Texto completo: 1 Coleções: 01-internacional Temas: Geral Base de dados: MEDLINE Assunto principal: Linfócitos T CD4-Positivos / Análise Serial de Tecidos Tipo de estudo: Diagnostic_studies Limite: Humans Idioma: En Revista: Anal Chem Ano de publicação: 2010 Tipo de documento: Article País de afiliação: Estados Unidos