Cystatins S and C in Human Whole Saliva and in Glandular Salivas in Periodontal Health and Disease

Abstract

Cystatins are inhibitors of cysteine proteinases and could play a protective and regulatory role under inflammatory conditions. Since total cystatin activity of whole saliva was increased in periodontal patients (Henskens et al., 1993), we wanted to investigate the types or origins of cystatins involved in this increase. Distinct types of cystatins were identified by isoelectric focusing and immunoblotting with specific antibodies against one of the salivary acidic isoforms, cystatin S. and the widely distributed basic cystatin C. Clarified human whole saliva (CHWS) of healthy subjects contained cystatin S, whereas cystatin C was barely detectable. In contrast, in CHWS of gingivitis and periodontitis patients, both cystatin C and S levels were higher. The origin of cystatin activity was investigated by collecting submandibular (SM), sublingual (SL), and parotid (PAR) saliva from seven subjects with mild gingivitis. Total cystatin activity was about five times higher in SM saliva than in PAR saliva. In SM and SL saliva, both cystatins S and C were demonstrated. In contrast, in PAR samples, solely cystatin C was detectable. The introduction of experimental gingivitis in one periodontally healthy subject resulted in the appearance of a cystatin C band in PAR saliva and in an increase of cystatins S and C in SM saliva. We conclude that the previously observed increase of cystatin activity in whole saliva in inflammatory periodontal disease is, at least in part, due to an increased glandular output of both the isoform cystatin S (pI 4.7) and the basic cystatin C (pI 9.0).

Innate Secretory Immunity in Response to Laboratory Stressors That Evoke Distinct Patterns of Cardiac Autonomic Activity

“…For the determination of cystatin S, a sandwich ELISA was used, as described by Henskens et al (44). Rabbit anti-human cystatin S polyclonal antiserum (purified immunoglobulin fraction) was used as a capture antibody, whereas a monoclonal antibody raised against cystatin S was used for detection (for details on the development of these antisera, see Refs.…”

Pathogenesis of periodontitis: a major arginine-specific cysteine proteinase from Porphyromonas gingivalis induces vascular permeability enhancement through activation of the kallikrein/kinin pathway.

“…Indeed, salivary protein and albumin concentrations of individuals with periodontitis, which are an indication of plasma leakage due to vascular permeability enhancement (VPE), are significantly increased compared to healthy subjects (24). These findings suggest that production of GCF is dependent on VPE induced at periodontitis sites, but little is known about the source or generation of VPE factor(s) and, more specifically, whether proteinase(s) of P. gingivalis are involved in their generation.…”

Antimicrobial Peptides in Saliva and Salivary Glands: Their Roles in the Oral Defense System

“…Cystatins (S, SA and SN) found in whole saliva are secreted from the SM-sublingual (SL) glands and, to a lesser extent, from the parotid. Henskens et al (17) reported that total cystatin activity in saliva is 5 times higher in SM saliva than in parotid saliva, and cystatins S and C are found in only SM and SL saliva.…”

“…In contrast, Henskens et al (20) showed that cystatin concentrations in whole saliva of periodontitis patients are significantly higher than those in healthy subjects. It has further been reported that an increase in cystatin levels in whole saliva of periodontal disease patients is in part due to increased salivary gland secretions of both isoform cystatin S and basic cystatin C (17). Cystatins and their synthetic derivatives are capable of blocking bacterial and viral replication and inhibiting the chemotactic response, and could modulate immune responses in mucosal tissue.…”