Abstract
The low frequency of T cells specific for given antigens makes the study of antigen-specific T cell responses difficult. The development of MHC class I and II tetramer staining techniques allows precise quantification and tracking of antigen-specific CD8+ and CD4+ T cell responses. Here, we describe a protocol for MHC class I and II tetramer staining of mouse T cells isolated from various tissues of mice infected with lymphocytic choriomeningitis virus (LCMV) or with murine cytomegalovirus (MCMV).
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Lau LL, Jamieson BD, Somasundaram T, Ahmed R (1994) Cytotoxic T-cell memory without antigen. Nature 369:648–652
Razvi ES, Welsh RM, McFarland HI (1995) In vivo state of antiviral CTL precursors. Characterization of a cycling cell population containing CTL precursors in immune mice. J Immunol 154:620–632
Selin LK, Nahill SR, Welsh RM (1994) Cross-reactivities in memory cytotoxic T lymphocyte recognition of heterologous viruses. J Exp Med 179:1933–1943
Tripp RA, Hou S, McMickle A, Houston J, Doherty PC (1995) Recruitment and proliferation of CD8+ T cells in respiratory virus infections. J Immunol 154:6013–6021
Butz EA, Bevan MJ (1998) Massive expansion of antigen-specific CD8+ T cells during an acute virus infection. Immunity 8:167–175
Marzo AL, Klonowski KD, Le Bon A, Borrow P, Tough DF, Lefrancois L (2005) Initial T cell frequency dictates memory CD8+ T cell lineage commitment. Nat Immunol 6:793–799
Altman JD, Moss PA, Goulder PJ, Barouch DH, McHeyzer-Williams MG, Bell JI, McMichael AJ, Davis MM (1996) Phenotypic analysis of antigen-specific T lymphocytes. Science 274:94–96
Murali-Krishna K, Altman JD, Suresh M, Sourdive DJ, Zajac AJ, Miller JD, Slansky J, Ahmed R (1998) Counting antigen-specific CD8 T cells: a reevaluation of bystander activation during viral infection. Immunity 8: 177–187
Fuller MJ, Zajac AJ (2003) Ablation of CD8 and CD4 T cell responses by high viral loads. J Immunol 170:477–486
Crawford F, Kozono H, White J, Marrack P, Kappler J (1998) Detection of antigen-specific T cells with multivalent soluble class II MHC covalent peptide complexes. Immunity 8: 675–682
Rees W, Bender J, Teague TK, Kedl RM, Crawford F, Marrack P, Kappler J (1999) An inverse relationship between T cell receptor affinity and antigen dose during CD4(+) T cell responses in vivo and in vitro. Proc Natl Acad Sci U S A 96:9781–9786
Wojciechowski S, Jordan MB, Zhu Y, White J, Zajac AJ, Hildeman DA (2006) Bim mediates apoptosis of CD127 (lo) effector T cells and limits T cell memory. Eur J Immunol 36: 1694–1706
Masopust D, Vezys V, Marzo AL, Lefrancois L (2001) Preferential localization of effector memory cells in nonlymphoid tissue. Science 291:2413–2417
Lin AA, Tripathi PK, Sholl A, Jordan MB, Hildeman DA (2009) Gamma interferon signaling in macrophage lineage cells regulates central nervous system inflammation and chemokine production. J Virol 83:8604–8615
Gairin JE, Mazarguil H, Hudrisier D, Oldstone MB (1995) Optimal lymphocytic choriomeningitis virus sequences restricted by H-2Db major histocompatibility complex class I molecules and presented to cytotoxic T lymphocytes. J Virol 69:2297–2305
Oxenius A, Bachmann MF, Ashton-Rickardt PG, Tonegawa S, Zinkernagel RM, Hengartner H (1995) Presentation of endogenous viral proteins in association with major histocompatibility complex class II: on the role of intracellular compartmentalization, invariant chain and the TAP transporter system. Eur J Immunol 25:3402–3411
Gold MC, Munks MW, Wagner M, Koszinowski UH, Hill AB, Fling SP (2002) The murine cytomegalovirus immunomodulatory gene m152 prevents recognition of infected cells by M45-specific CTL but does not alter the immunodominance of the M45-specific CD8 T cell response in vivo. J Immunol 169:359–365
Arens R, Wang P, Sidney J, Loewendorf A, Sette A, Schoenberger SP, Peters B, Benedict CA (2008) Cutting edge: murine cytomegalovirus induces a polyfunctional CD4 T cell response. J Immunol 180:6472–6476
Tabeta K, Georgel P, Janssen E, Du X, Hoebe K, Crozat K, Mudd S, Shamel L, Sovath S, Goode J, Alexopoulou L, Flavell RA, Beutler B (2004) Toll-like receptors 9 and 3 as essential components of innate immune defense against mouse cytomegalovirus infection. Proc Natl Acad Sci U S A 101:3516–3521
Orange JS, Wang B, Terhorst C, Biron CA (1995) Requirement for natural killer cell-produced interferon gamma in defense against murine cytomegalovirus infection and enhancement of this defense pathway by interleukin 12 administration. J Exp Med 182:1045–1056
Acknowledgments
This work was supported by Public Health Service Grants AI057753, DK081175 (D.A.H.).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media New York
About this protocol
Cite this protocol
Kurtulus, S., Hildeman, D. (2013). Assessment of CD4+ and CD8+ T Cell Responses Using MHC Class I and II Tetramers. In: Snow, A., Lenardo, M. (eds) Immune Homeostasis. Methods in Molecular Biology, vol 979. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-290-2_8
Download citation
DOI: https://doi.org/10.1007/978-1-62703-290-2_8
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-289-6
Online ISBN: 978-1-62703-290-2
eBook Packages: Springer Protocols