Detection and Characterization of Virus-Specific CD8+ T Cells Using the Tetramer Approach

  • Xiao-Song He
  • Harry B. Greenberg
Part of the Methods in Molecular Medicine™ book series (MIMM, volume 96)


Hepatitis B virus (HBV)-specific CD8+ T cells, or cytotoxic T lymphocytes (CTLs), are believed to play an important role in the control of infection and development of liver injury (1). Therefore the quantitative and qualitative analyses of such cells is important for understanding the mechanisms underlying the pathogenesis associated with the infection as well as the clearance or persistence of the infecting virus. Traditionally, HBV-specific CD8+ T cells were detected by cytotoxicity assay, while the frequency of such cells was estimated by limiting dilution assay (LDA). Both assays are based on the killing activity of CTLs. The number of virus-specific CTLs in clinical specimens (blood or liver samples) is usually very small. Therefore these assays require extensive in vitro expansion of virus-specific CD8+ T cells to generate CTL lines or clones, which is time consuming and labor intensive. As such assays can detect only CD8+ T cells that expand in vitro and kill specific target cells, they may not represent the characteristics of the overall CD8+ T cell responses against the virus.


Phosphatidyl Ethanolamine Tetramer Staining PBMC Sample Limit Dilution Assay Tetramer Assay 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


  1. 1.
    Guidotti, L. G. and Chisari, F. V. (2001) Noncytolytic control of viral infections by the innate and adaptive immune response. Annu. Rev. Immunol. 19, 65–91.PubMedCrossRefGoogle Scholar
  2. 2.
    Altman, J. D., Moss, P., Goulder, P., et al. (1996) Phenotypic analysis of antigen-specific T lymphocytes (published erratum appears in Science [1998] 280, 1821). Science 274, 94–96.PubMedCrossRefGoogle Scholar
  3. 3.
    He, X. S., Rehermann, B., Lopez-Labrador, F. X., et al. (1999) Quantitative analysis of hepatitis C virus-specific CD8(+) T cells in peripheral blood and liver using peptide-MHC tetramers. Proc. Natl. Acad. Sci. USA 96, 5692–5697.PubMedCrossRefGoogle Scholar
  4. 4.
    He, X. S., Rehermann, B., Boisvert, J., et al. (2001) Direct functional analysis of epitope-specific CD8+ T cells in peripheral blood. Viral Immunol 14, 59–69.PubMedCrossRefGoogle Scholar
  5. 5.
    Lee, P. P., Yee, C., Savage, P. A., et al., (1999) Characterization of circulating T cells specific for tumor-associated antigens in melanoma patients. Nat. Med. 5, 677–685.PubMedCrossRefGoogle Scholar
  6. 6.
    Maini, M. K., Boni, C., Ogg, G. S., et al. (1999) Direct ex vivo analysis of hepatitis B virus-specific CD8(+) T cells associated with the control of infection. Gastroenterology 117, 1386–1396.PubMedCrossRefGoogle Scholar
  7. 7.
    Boni, C., Penna, A., Ogg, G. S., et al. (2001) Lamivudine treatment can overcome cytotoxic T-cell hyporesponsiveness in chronic hepatitis B: new perspectives for immune therapy. Hepatology 33, 963–971.PubMedCrossRefGoogle Scholar
  8. 8.
    Maini, M. K., Boni, C., Lee, C. K., et al., (2000) The role of virus-specific CD8(+) cells in liver damage and viral control during persistent hepatitis B virus infection. J. Exp. Med. 191, 1269–1280.PubMedCrossRefGoogle Scholar
  9. 9.
    Doherty, P. C. (1998) The new numerology of immunity mediated by virus-specific CD8(+) T cells. Curr. Opin. Microbiol. 1, 419–422.PubMedCrossRefGoogle Scholar
  10. 10.
    McMichael, A. J. and O’Callaghan, C. A. (1998) A new look at T cells. J. Exp. Med. 187, 1367–1371.PubMedCrossRefGoogle Scholar
  11. 11.
    Valitutti, S., Muller, S., Cella, M., Padovan, E., and Lanzavecchia, A. (1995) Serial triggering of many T-cell receptors by a few peptide-MHC complexes (see comments). Nature 375, 148–151.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc. 2004

Authors and Affiliations

  • Xiao-Song He
    • 1
  • Harry B. Greenberg
    • 1
  1. 1.Division of Gastroenterology and HepatologyStanford University School of Medicine and Veterans Administration Medical CenterPalo Alto

Personalised recommendations