Single-strand conformation polymorphism analysis for the diagnosis of T-cell clonality in periodontal disease

Yamazaki, K.; Ito, H.

Methods in Molecular Biology 666: 359-372

2010


ISSN/ISBN: 1940-6029
PMID: 20717795
DOI: 10.1007/978-1-60761-820-1_22
Accession: 055799981

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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.