title: Intracellular localization of Treponema denticola chymotrypsin-like proteinase in chronic periodontitis.
abstract: Treponema denticola is an important periodontal pathogen capable of tissue invasion. Its chymotrypsin-like proteinase (CTLP) can degrade a number of basement membrane components in vitro, thus suggesting a contribution to tissue invasion by the spirochete. The aim of this study was to analyze the localization of CTLP in chronic periodontitis tissues ex vivo. A polyclonal antibody specific to T. denticola cell-bound CTLP was used to detect the spirochetes in the gingival tissues of patients with moderate to severe chronic periodontitis (n=25) by immunohistochemistry and periodic acid-Schiff staining (PAS). The presence of T. denticola in the periodontal tissue samples was analyzed by PCR. Periodontal tissue samples of 12 of the 25 patients were found to be positive for T. denticola by PCR. Moreover, CTLP could be detected in the periodontal tissues of all these patients by immunohistochemistry. In the epithelium, the CTLP was mostly intracellular. Typically, the positive staining could be seen throughout the whole depth of the epithelium. When detected extracellularly, CTLP was localized mainly as granular deposits. The connective tissue stained diffusely positive in four cases. The positive staining co-localized with the PAS stain in nine cases. T. denticola and its CTLP could be detected in diseased human periodontium both intra- and extracellularly. The granular staining pattern was suggestive of the presence of T. denticola bacteria, whereas the more diffused staining pattern was indicative of the recent presence of the bacterium and shedding of the cell-bound proteinase.
authors.affiliation: Department of Oral and Maxillofacial Diseases, Helsinki University Central Hospital, Helsinki, Finland.
authors.forename: Emilia
authors.initials: E
authors.lastname: Marttila
authors.affiliation: Institute of Dentistry, University of Helsinki, Helsinki, Finland.
authors.forename: Anne
authors.initials: A
authors.lastname: Järvensivu
authors.affiliation: Department of Oral and Maxillofacial Diseases, Helsinki University Central Hospital, Helsinki, Finland ; Institute of Dentistry, University of Helsinki, Helsinki, Finland ; Division of Periodontology, Department of Dental Medicine, Karolinska Institute, Huddinge, Sweden.
authors.forename: Timo
authors.initials: T
authors.lastname: Sorsa
authors.affiliation: Faculte de Medecine Dentaire, Groupe de Recherche en Ecologie Buccale, Universite Laval, Quebec, Canada.
authors.forename: Daniel
authors.initials: D
authors.lastname: Grenier
authors.affiliation: Mycology Reference Centre, University Hospital of South Manchester, Manchester Academic Health Science Centre, School of Translational Medicine, University of Manchester, Manchester, UK.
authors.forename: Malcolm
authors.initials: M
authors.lastname: Richardson
authors.affiliation: Institute of Dentistry, University of Helsinki, Helsinki, Finland.
authors.forename: Kirsti
authors.initials: K
authors.lastname: Kari
authors.affiliation: Institute of Dentistry, University of Helsinki, Helsinki, Finland.
authors.forename: Taina
authors.initials: T
authors.lastname: Tervahartiala
authors.affiliation: Manchester Academic Health Science Centre, NIHR Translational Research Facility, Translational Research Facility, Institute of Inflammation and Repair, University of Manchester and University Hospital of South Manchester, Manchester, UK.
authors.forename: Riina
authors.initials: R
authors.lastname: Rautemaa
keywords: dentilisin; major outer sheath protein; spirochetes