Ultra-sensitive class I tetramer analysis reveals previously undetectable populations of antiviral CD8+ T cells

Eur J Immunol. 2004 Jun;34(6):1570-7. doi: 10.1002/eji.200424898.


A major breakthrough in cellular immunology has been the development of HLA class I tetramers to analyze CD8(+) T cell responses. However, in many situations, including persistent virus infection, specific T cell responses are rarely detected using this technology. This raises the question of whether such responses are 'deleted' (or 'exhausted') or present below the conventional detection limit for class I tetramer staining. In particular, persistent hepatitis C virus (HCV) infection is characterized by very weak or apparently absent specific CD8(+) T cell responses, even though they are readily detectable in acute disease. Therefore, we assessed the use of anti-PE-labeled magnetic beads to enrich tetramer-positive HCV-specific T cells and identify previously undetectable populations. Using the enrichment technique, HCV-specific T cells could be detected in the majority of infected individuals, whereas these responses were not detected using conventional tetramer staining (8/15 vs. 1/15; p=0.01). Magnetic enrichment could reliably detect very rare HCV-specific responses at frequencies of >0.0011% of CD8(+) T cells (approximately 1/million PBMC), and phenotypic analysis of these rare populations was possible. Therefore, this direct ex vivo technique revealed the persistence of very low frequencies of virus-specific CD8(+) T cells during chronic virus infection and is readily transferable to the study of other viral, self- or tumor-specific T cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, Neoplasm
  • CD8-Positive T-Lymphocytes / immunology*
  • CD8-Positive T-Lymphocytes / virology
  • HLA-A2 Antigen / immunology*
  • Hepacivirus / immunology*
  • Hepatitis C Antigens / immunology
  • Hepatitis C Antigens / isolation & purification
  • Hepatitis C, Chronic / immunology*
  • Hepatitis C, Chronic / virology
  • Humans
  • Immunomagnetic Separation / methods
  • MART-1 Antigen
  • Neoplasm Proteins / immunology
  • Phycoerythrin / immunology


  • Antigens, Neoplasm
  • HLA-A2 Antigen
  • Hepatitis C Antigens
  • MART-1 Antigen
  • MLANA protein, human
  • Neoplasm Proteins
  • Phycoerythrin