Skip to main content

Quantification of a Selective Expansion of T Cell Receptor Vβ by Superantigen Using Real-Time PCR

  • Protocol
  • First Online:
Superantigens

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1396))

Abstract

Selective expansion of T cells bearing specific T cell receptor Vβ segments is a hallmark of superantigens. Analyzing Vβ specificity of superantigens is important for characterizing newly discovered superantigens and understanding differential T cell responses to each toxin. Here, we describe a real-time PCR method using SYBR green I and primers specific to Cβ and Vβ genes for an absolute quantification. The established method was applied to quantify a selective expansion of T cell receptor Vβ expansion by superantigens and generated accurate, reproducible, and comparable results.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Davis MM, Boniface JJ, Reich Z, Lyons D, Hampl J, Arden B, Chien Y (1998) Ligand recognition by alpha beta T cell receptors. Annu Rev Immunol 16:523–544

    Article  CAS  PubMed  Google Scholar 

  2. Rowen L, Koop BF, Hood L (1996) The complete 685-kilobase DNA sequence of the human beta T cell receptor locus. Science 272:1755–1762

    Article  CAS  PubMed  Google Scholar 

  3. Davis MM, Bjorkman PJ (1988) T-cell antigen receptor genes and T-cell recognition. Nature 334:395–402

    Article  CAS  PubMed  Google Scholar 

  4. Behlke MA, Spinella DG, Chou HS, Sha W, Hartl DL, Loh DY (1985) T-cell receptor beta-chain expression: dependence on relatively few variable region genes. Science 229:566–570

    Article  CAS  PubMed  Google Scholar 

  5. Lefranc M, Lefranc G (2001) The T cell receptor. Academic, New York

    Google Scholar 

  6. Arden B, Clark SP, Kabelitz D, Mak TW (1995) Human T-cell receptor variable gene segment families. Immunogenetics 42:455–500

    CAS  PubMed  Google Scholar 

  7. Seo KS, Bohach GA (2007) Staphylcoccus aureus. In: Doyle MM, Beucaht LR (eds) Food microbiology: fundamentals and Frontiers, 3rd edn. ASM Press, Washington, DC, pp 493–518

    Chapter  Google Scholar 

  8. Bohach GA (2006) Staphylococcus aureus Exotoxins. In: Fischetti VA, Novick RP, Ferretti JJ, Portnoy DA, Rood JI (eds) Gram-positive pathogens, 2nd edn. ASM Press, Washington, DC, pp 464–477

    Chapter  Google Scholar 

  9. Deringer JR, Ely RJ, Stauffacher CV, Bohach GA (1996) Subtype-specific interactions of type C staphylococcal enterotoxins with the T-cell receptor. Mol Microbiol 22:523–534

    Article  CAS  PubMed  Google Scholar 

  10. Pilch H, Hohn H, Freitag K, Neukirch C, Necker A, Haddad P, Tanner B, Knapstein PG, Maeurer MJ (2002) Improved assessment of T-cell receptor (TCR) VB repertoire in clinical specimens: combination of TCR-CDR3 spectratyping with flow cytometry-based TCR VB frequency analysis. Clin Diagn Lab Immunol 9:257–266

    CAS  PubMed  PubMed Central  Google Scholar 

  11. Bercovici N, Duffour MT, Agrawal S, Salcedo M, Abastado JP (2000) New methods for assessing T-cell responses. Clin Diagn Lab Immunol 7:859–864

    CAS  PubMed  PubMed Central  Google Scholar 

  12. Seo KS, Park JY, Terman DS, Bohach GA (2010) A quantitative real time PCR method to analyze T cell receptor Vbeta subgroup expansion by staphylococcal superantigens. J Transl Med 8:2. doi:10.1186/1479-5876-8-2

    Article  PubMed  PubMed Central  Google Scholar 

  13. Sambrook J, Fritsch EF, Maniatis T (1989) Concentration of DNA solution. In: Nolan C (ed) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, New York, p Appendix C1

    Google Scholar 

  14. Yin JL, Shackel NA, Zekry A, McGuinness PH, Richards C, Putten KV, McCaughan GW, Eris JM, Bishop GA (2001) Real-time reverse transcriptase-polymerase chain reaction (RT-PCR) for measurement of cytokine and growth factor mRNA expression with fluorogenic probes or SYBR Green I. Immunol Cell Biol 79:213–221

    Article  CAS  PubMed  Google Scholar 

  15. Lamphear JG, Mollick JA, Reda KB, Rich RR (1996) Residues near the amino and carboxyl termini of staphylococcal enterotoxin E independently mediate TCR V beta-specific interactions. J Immunol 156:2178–2185

    CAS  PubMed  Google Scholar 

  16. Choi YW, Kotzin B, Herron L, Callahan J, Marrack P, Kappler J (1989) Interaction of Staphylococcus aureus toxin “superantigens” with human T cells. Proc Natl Acad Sci U S A 86:8941–8945

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Jarraud S, Peyrat MA, Lim A, Tristan A, Bes M, Mougel C, Etienne J, Vandenesch F, Bonneville M, Lina G (2001) egc, a highly prevalent operon of enterotoxin gene, forms a putative nursery of superantigens in Staphylococcus aureus. J Immunol 166:669–677

    Article  CAS  PubMed  Google Scholar 

  18. Kappler J, Kotzin B, Herron L, Gelfand EW, Bigler RD, Boylston A, Carrel S, Posnett DN, Choi Y, Marrack P (1989) V beta-specific stimulation of human T cells by staphylococcal toxins. Science 244:811–813

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Keun Seok Seo .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2016 Springer Science+Business Media New York

About this protocol

Cite this protocol

Park, J.Y., Seo, K.S. (2016). Quantification of a Selective Expansion of T Cell Receptor Vβ by Superantigen Using Real-Time PCR. In: Brosnahan, A. (eds) Superantigens. Methods in Molecular Biology, vol 1396. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3344-0_15

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-3344-0_15

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3342-6

  • Online ISBN: 978-1-4939-3344-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics