On-chip activation and subsequent detection of individual antigen-specific T cells

Forskningsoutput: TidskriftsbidragLetter

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.

Detaljer

Författare
  • Qing Song
  • Qing Han
  • Elizabeth M Bradshaw
  • Sally C Kent
  • Khadir Raddassi
  • Björn Nilsson
  • Gerald T Nepom
  • David A Hafler
  • J Christopher Love
Externa organisationer
  • Broad Institute
Forskningsområden

Ämnesklassifikation (UKÄ) – OBLIGATORISK

  • Medicinsk genetik

Nyckelord

Originalspråkengelska
Sidor (från-till)473-7
Antal sidor5
TidskriftAnalytical Chemistry
Volym82
Utgivningsnummer2
StatusPublished - 2010 jan 15
PublikationskategoriForskning
Peer review utfördJa
Externt publiceradJa