Skip to main content

Single-Strand Conformation Polymorphism Analysis for the Diagnosis of T-Cell Clonality in Periodontal Disease

  • Protocol
  • First Online:
Oral Biology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 666))

Abstract

T cells recognize antigens via the T-cell receptor (TCR). Diversity in antigen recognition by T cells is generated in part by the recombination of V, (D), J, and C segments of the TCR. It is further enhanced by the N region, in addition to non-germline-encoded nucleotides at the V–(D)–J junction. It is generally believed that each T cell bears a distinct clonotype of TCR and that each clonotype is responsible for an antigen-specific T-cell response. T-cell clonal expansion has been detected in the peripheral blood or the disease-affected sites in patients with infections, autoimmune diseases, malignancy, and post-transplantation complications. Since antigen stimulation of T cells induces the proliferation of specific T cells, clonal T-cell expansion is considered to be a result of an antigen-specific immune response. For the analysis of such antigen-specific T cells, it is common to use their specific antigens if they are known. However, there are many diseases, such as periodontal diseases, in which there are a number of putative pathogenic antigens involved. In these circumstances, the detection of clonally expanded T cells is an effective method to evaluate whether antigen-specific immune responses are involved, since only a few clonally expanded T cells are detected in healthy individuals. In addition, the characterization of any clonally expanded T cells that are detected would further promote the understanding of the disease mechanisms. By using single-strand conformation polymorphism (SSCP) analysis, we demonstrated that oligoclonal T-cell accumulation was present in periodontitis lesions, in contrast to a heterogeneous T-cell population in the peripheral blood. SSCP is a powerful tool for analyzing specific T-cell responses both in vitro and in vivo.

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 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
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. Seymour, G. J. (1991) Importance of the host response in the periodontium. J. Clin. Periodontol. 18, 421–426.

    Article  PubMed  CAS  Google Scholar 

  2. Matis, L. A. (1990) The molecular basis of T-cell specificity. Annu. Rev. Immunol. 8, 65–82.

    Article  PubMed  CAS  Google Scholar 

  3. Mathur, A., Michalowicz, B., Yang, C., and Aeppli, D. (1995) Influence of periodontal bacteria and disease status on V beta expression in T cells. J. Periodontal. Res. 30, 369–373.

    Article  PubMed  CAS  Google Scholar 

  4. Nakajima, T., Yamazaki, K., and Hara, K. (1996) Biased T cell receptor V gene usage in tissues with periodontal disease. J. Periodontal. Res. 31, 2–10.

    Article  PubMed  CAS  Google Scholar 

  5. Wassenaar, A., Reinhardus, C., Thepen, T., Abraham-Inpijn, L., and Kievits, F. (1995) Cloning, characterization, and antigen specificity of T-lymphocyte subsets extracted from gingival tissue of chronic adult periodontitis patients. Infect. Immun. 63, 2147–2153.

    PubMed  CAS  Google Scholar 

  6. Masuko, K., Kato, T., Ikeda, Y., Okubo, M., Mizushima, Y., Nishioka, K. et al. (1994) Dynamic changes of accumulated T cell clonotypes during antigenic stimulation in vivo and in vitro. Int. Immunol. 6, 1959–1966.

    Article  PubMed  CAS  Google Scholar 

  7. Orita, M., Iwahana, H., Kanazawa, H., Hayashi, K., and Sekiya, T. (1989) Detection of polymorphisms of human DNA by gel electrophoresis as single-strand conformation polymorphisms. Proc. Natl. Acad. Sci. USA. 86, 2766–2770.

    Article  PubMed  CAS  Google Scholar 

  8. Yamamoto, K., Sakoda, H., Nakajima, T., Kato, T., Okubo, M., Dohi, M. et al. (1992) Accumulation of multiple T cell clonotypes in the synovial lesions of patients with rheumatoid arthritis revealed by a novel clonality analysis. Int. Immunol. 4, 1219–1223.

    Article  PubMed  CAS  Google Scholar 

  9. Yamazaki, K., and Nakajima, T. (2004) Antigen specificity and T-cell clonality in periodontal disease. Periodontol. 2000. 35, 75–100.

    Article  PubMed  Google Scholar 

  10. Itoh, H., Ohsawa, Y., Yoshie, H., and Yamazaki, K. (2002) Oligoclonal accumulations of T-cell clones in gingivitis and periodontitis lesions. Oral Microbiol. Immunol. 17, 324–329.

    Article  PubMed  CAS  Google Scholar 

  11. Yamazaki, K., Nakajima, T., Ohsawa, Y., Tabeta, K., Yoshie, H., Sakurai, K. et al. (2000) Selective expansion of T cells in gingival lesions of patients with chronic inflammatory periodontal disease. Clin. Exp. Immunol. 120, 154–161.

    Article  PubMed  CAS  Google Scholar 

  12. Ohsawa, Y., Yamazaki, K., Nakajima, T., and Hara, K. (2000) Clonal accumulation of T cells bearing V beta 6 T-cell receptor in chronic inflammatory periodontal disease. Oral. Microbiol. Immunol. 15, 211–217.

    Article  PubMed  CAS  Google Scholar 

  13. Nakajima, T., Yamazaki, K., Sakurai, K., Gemmell, E., Seymour, G. J., and Hara, K. (1998) Detection of clonotypic changes of T cells after stimulation with Porphyromonas gingivalis. Oral Microbiol. Immunol. 13, 238–245.

    Article  PubMed  CAS  Google Scholar 

  14. Yamazaki, K., Ohsawa, Y., Tabeta, K., Ito, H., Ueki, K., Oda, T. et al. (2002) Accumulation of human heat shock protein 60-reactive T cells in the gingival tissues of periodontitis patients. Infect. Immun. 70, 2492–2501.

    Article  PubMed  CAS  Google Scholar 

  15. Yamazaki, K., Ohsawa, Y., and Yoshie, H. (2001) Elevated proportion of natural killer T cells in periodontitis lesions: a common feature of chronic inflammatory diseases. Am. J. Pathol. 158, 1391–1398.

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

The authors would like to thank Drs. Yutaka Ohsawa and Takako Nakajima for their technical assistance. This work was supported by Grants-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science, and Technology of Japan (19390536, 20659325) and the Promotion of Niigata University Research Project. Pacific Edit reviewed the manuscript prior to submission.

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2010 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Yamazaki, K., Ito, H. (2010). Single-Strand Conformation Polymorphism Analysis for the Diagnosis of T-Cell Clonality in Periodontal Disease. In: Seymour, G., Cullinan, M., Heng, N. (eds) Oral Biology. Methods in Molecular Biology, vol 666. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-820-1_22

Download citation

  • DOI: https://doi.org/10.1007/978-1-60761-820-1_22

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-60761-819-5

  • Online ISBN: 978-1-60761-820-1

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics