Regulatory T Cells pp 45-54

Part of the Methods in Molecular Biology book series (MIMB, volume 707)

In Vitro and In Vivo Analyses of Regulatory T Cell Suppression of CD8+ T Cells

Protocol

Abstract

The study of regulatory T cells (Treg) requires methods for both in vivo and in vitro analyses, both of which have different limitations, but which complement each other to give a more complete picture of physiological function than either method alone. Our analyses have focused on Treg-mediated suppression of CD8+ T cells, and in particular Tregs induced by viral infection. One of the unique characteristics of virus-induced Tregs is that they can suppress CD8+ T cell function in vitro without the requirement for additional stimulation. This ability correlates with their suppressive capacity and activated status in vivo. Interestingly, while virus-induced Tregs suppress CD8+ T cell function in vitro and in vivo, they do not suppress proliferation unless they are further activated in vitro.

Key words

Regulatory T cells CD8+ T cells 

References

  1. 1.
    Hasenkrug, K. J. and Dittmer, U. (2007) Immune control and prevention of chronic Friend retrovirus infection. Front. Biosci. 12, 1544–1551.PubMedCrossRefGoogle Scholar
  2. 2.
    Hasenkrug, K. J. and Chesebro, B. (1997) Immunity to retroviral infection: the Friend virus model. Proc. Natl. Acad. Sci. USA 94, 7811–7816.PubMedCrossRefGoogle Scholar
  3. 3.
    Hasenkrug, K. J., Brooks, D. M. and Dittmer, U. (1998) Critical role for CD4+ T cells in controlling retrovirus replication and spread in persistently infected mice. J. Virol. 72, 6559–6564.PubMedGoogle Scholar
  4. 4.
    Dittmer, U., He, H., Messer, R. J., et al. (2004) Functional impairment of CD8(+) T cells by regulatory T cells during persistent retroviral infection. Immunity 20, 293–303.PubMedCrossRefGoogle Scholar
  5. 5.
    Hasenkrug, K. J. (1999) Lymphocyte deficiencies increase susceptibility to Friend virus-induced erythroleukemia in Fv-2 genetically resistant mice. J. Virol. 73, 6468–6473.PubMedGoogle Scholar
  6. 6.
    Iwashiro, M., Messer, R. J., Peterson, K. E., Stromnes, I. M., Sugie, T. and Hasenkrug, K. J. (2001) Immunosuppression by CD4+ regulatory T cells induced by chronic retroviral infection. Proc. Natl. Acad. Sci. USA 98, 9226–9230.PubMedCrossRefGoogle Scholar
  7. 7.
    Sakaguchi, S., Sakaguchi, N., Asano, M., Itoh, M. and Toda, M. (1995) Immunologic self-tolerance maintained by activated T cells expressing IL-2 receptor alpha-chains (CD25). Breakdown of a single mechanism of self-tolerance causes various autoimmune diseases. J. Immunol. 155, 1151–1164.PubMedGoogle Scholar
  8. 8.
    Myers, L., Messer, R. J., Carmody, A. B. and Hasenkrug, K. J. (2009) Tissue-specific abundance of regulatory T cells correlates with CD8+ T cell dysfunction and chronic retrovirus loads. J. Immunol. 183, 1636–1643.PubMedCrossRefGoogle Scholar
  9. 9.
    Robertson, S. J., Messer, R. J., Carmody, A. B. and Hasenkrug, K. J. (2006) In vitro suppression of CD8+ T cell function by Friend virus-induced regulatory T cells. J. Immunol. 176, 3342–3349.PubMedGoogle Scholar
  10. 10.
    Shevach, E. M. (2002) CD4+ CD25+ suppressor T cells: more questions than answers. Nat. Rev. Immunol. 2, 389–400.PubMedGoogle Scholar
  11. 11.
    Von Boehmer, H. (2005) Mechanisms of suppression by suppressor T cells. Nat. Immunol. 6, 338–344.CrossRefGoogle Scholar
  12. 12.
    Bettelli, E., Carrier, Y., Gao, W., et al. (2006) Reciprocal developmental pathways for the generation of pathogenic effector TH17 and regulatory T cells. Nature 441, 235–238.PubMedCrossRefGoogle Scholar
  13. 13.
    Zelinskyy, G., Kraft, A. R., Schimmer, S., Arndt, T. and Dittmer, U. (2006) Kinetics of CD8+ effector T cell responses and induced CD4+ regulatory T cell responses during Friend retrovirus infection. Eur. J. Immunol. 36, 2658–2670.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.Laboratory of Persistent Viral Diseases, Rocky Mountain LaboratoriesNational Institute of Allergy and Infectious Diseases, National Institutes of HealthHamiltonUSA

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