Detection of clonotypic changes of T cells after stimulation with Porphyromonas gingivalis.

Abstract

The aim of this study was to investigate whether Porphyromonas gingivalis stimulation would induce a selective activation and expansion of a limited T-cell receptor V beta repertoire or T-cell clonotype. Using samples from patients with chronic inflammatory periodontal diseases, we examined TCRBV gene usage and T-cell clonotypes of peripheral blood mononuclear cells incubated in the presence or absence of P. gingivalis outer membrane through a combination of reverse transcription polymerase chain reaction (RT-PCR) and subsequent single strand conformation polymorphism (SSCP) analysis. There was no difference in the mean expression for most BV families with or without P. gingivalis outer membrane in the culture. However, in individual cases, a few BV gene families did become overexpressed or underexpressed following stimulation, although a consistent pattern did not emerge. SSCP analysis showed that several new distinct bands appeared after stimulation, indicating distinct clonal accumulations, although the number of distinct bands decreased in most cases. These data suggest that clonotypic change occurred following stimulation with P. gingivalis outer membrane. Furthermore, the possibility of superantigen stimulation by P. gingivalis is unlikely to be due to the small change in BV gene usage and clonal T-cell accumulations with P. gingivalis outer membrane stimulation, as evidenced by SSCP. Thus, RT-PCR and SSCP analysis is useful in evaluating the host response to periodontopathic antigens.