Indicators of salivary gland inflammation in primary Sjögren's syndrome

The aim of this study was to establish additional indicators in saliva and plasma which are associated with salivary gland inflammation in patients with primary Sjögren's syndrome (SS). ELISA assays were used to determine the concentrations of sICAM-1, sVCAM-1, sIL-2Rz, IgA, IgG, calprotectin and albumin in parotid saliva, whole saliva and plasma samples. Soluble ICAM-1 was present in whole and parotid saliva samples from primary SS patients. Soluble VCAM-1 and sIL-IRx could not be detected in salivary samples from either primary SS or control subjects. IgA, IgG, calprotectin and albumin concentrations were higher in both whole and parotid saliva in the patient group compared with the control group. The results showed increased levels of calprotectin in all saliva samples compared to plasma, suggesting that calprotectin may be locally produced. Increased plasma values of sICAM-1. sVCAM-1, SIL-2Rα, IgA, IgG and calprotectin were detected in primary SS patients when compared to controls. The output/min of IgA. IgG, calprotectin and albumin was decreased in SS patients. Plasma levels of various proteins could offer information concerning glandular and extraglandular inflammatory processes. However, salivary levels of these proteins (particularly sICAM-1) tend to reflect more the local inflammatory activity, providing a convenient and non-invasive tool for diagnosis.     

Protein composition of whole and parotid saliva in healthy and periodontitis subjects

Cystatins are physiological inhibitors of cysteine proteinases and they are widely distributed in human tissues and body fluids including saliva. We previously reported an increased cystatin activity in whole saliva of gingivitis and periodontitis subjects. Based on this result we decided to investigate the type and origin of cystatins involved in this increased cystatin activity by collecting both whole and parotid saliva of 25 healthy and 30 periodontitis subjects. Saliva samples were quantified for cystatins S and C by enzyme-linked immunosorbent assay and cystatin activities were measured toward papain. Besides, three other salivary proteins were determined: the plasma protein albumin, the typical parotid derived amylase and the salivary immunoglobulin IgA. The present investigation shows that levels of total protein and cystatin activity as well as the levels of glandular derived proteins amylase and cystatin C were significantly higher in whole and parotid saliva of subjects with periodontitis than in healthy controls. Cystatin S, the major salivary cystatin. however was higher in the whole saliva of the healthy group. Whole saliva concentrations of albumin and IgA, originating from sources other than the glandular cells, were not different between healthy and periodontitis subjects and were also not correlated with the typical salivary gland proteins. In conclusion, this study provides additional evidence that the human salivary glands may respond to an inflammatory disease of the oral cavity, periodontitis, by enhanced synthesis of some acinar proteins.     

Minor salivary gland immunohistology in the diagnosis of primary Sjögren's syndrome

Background:  Focal lymphocytic infiltrates of minor salivary glands are considered target-organ related signs of Sjögren's syndrome. The percentages of plasma cells expressing IgA, IgG and IgM in minor salivary gland biopsies have also been suggested as useful in establishing a diagnosis of Sjögren's syndrome, and this study aimed at evaluating this method.
Methods:  All biopsies from patients under investigation for Sjögren's syndrome ( n  = 210) at our department during 4 years were analyzed for IgA, IgG and IgM producing cells by immunohistochemistry, and related to Sjögren classification parameters.
Results:  A focus score ≥1 was observed in 67/210 patients and the frequency of IgA producing cells was <70% in 42/210 patients. Sufficient clinical data for classification of disease were available for 57/210 patients. Patients were classified as having primary Sjögren's syndrome (pSS) ( n  = 9), secondary Sjögren's syndrome (sSS) ( n  = 12) or non-Sjögren's syndrome (non-SS) ( n  = 36). IgA expressing cells were significantly decreased ( P  < 0.01) and IgG expressing cells significantly increased ( P  < 0.02) in patients with pSS compared to non-SS. Also, increased numbers of salivary gland IgG producing plasma cells correlated with increased IgG serum levels ( P  < 0.001). However, there was no significant difference between sSS and non-SS with regard to IgA, IgG or IgM expressing cells in the glands.
Conclusions:  Our results support previous reports indicating the relevance of quantitative evaluation of Ig isotype expression in plasma cells in the clinical investigation of Sjögren's syndrome and further indicate a difference in plasma cell populations between pSS and sSS.     

Salivary protein composition in epileptic patients on different medications

Several salivary proteins were assayed in saliva from epileptic patients who were using different anti-epileptic drugs, viz. phenytoin, valproate and carbamazepine, and were compared with levels in unmedicated healthy control subjects. Flow rate and pH of the patient groups were not different from the controls. In all patient groups the specific amylase activity was increased up to twofold. In the phenytoin group only, the salivary IgA concentration was strongly reduced. Levels of salivary cystatin C were similar among all patient groups studied, and were not different from those of the control group. In contrast, the absolute and relative concentrations of cystatin S were diminished, particularly in patients using either valproate or phenytoin. These data suggest that use of anti-epileptic drugs over long periods may result in decreased levels of several salivary proteins such as slgA and cystatins. which are involved in the protection of the oral cavity against microbial infections.     

Correlations between total protein, lysozyme, immunoglobulins, amylase, and albumin in stimulated whole saliva during daytime

The correlations between salivary proteins and the daytime variations are not known. The present study investigated the within-subject variation of correlations and concentrations between lysozyme, IgA, IgG, IgM, albumin, amylase, and total protein in stimulated whole saliva of healthy adults in the course of a 12-h period. After several practise sessions, unstimulated and stimulated whole saliva samples were collected five times daily (at 8 a.m., 11 a.m., 2 p.m., 5 p.m., and 8 p.m.) from 30 healthy university students. Flow rate and total protein concentration were used as covariates, and gender as a between-subject factor in the MANOVA analysis. After this adjustment, there was significant within-subject variation in salivary IgA (P < 0.001), albumin (P < 0.01), amylase (P < 0.05), and total protein (P < 0.001) concentrations. Total protein correlated significantly with amylase albumin and IgA through different samplings. In addition, IgG correlated with albumin and lysozyme in the course of 12 h. On the whole, the correlations between variables remained stable during repeated samplings. In addition, rankings of subjects for the variables tended to be maintained across different samplings (P < 0.001). However, the observed within-subject variations in salivary IgA, albumin, amylase, and total protein concentrations suggest that these proteins are subject to short-term variation.     

Cytokines in Sjögren's syndrome

Cytokines play a central role in the regulation of immunity and are often found to be deregulated in autoimmune diseases. Sjögren's syndrome is a chronic autoimmune disease characterized by inflammation and loss of secretory function of the salivary and lachrymal glands. This review highlights the current knowledge of the expression and the function of pro- and anti-inflammatory cytokines both locally and systemically in Sjögren's syndrome patients. In the salivary glands, saliva and serum of these patients, many pro-inflammatory cytokines are upregulated. Concomitantly, most anti-inflammatory cytokines are not detectable or are expressed at low levels. Besides a role in inflammation, cytokines are also thought to be involved in salivary gland dysfunction by directly interfering with the epithelial cells in the glands. Future research on the role of novel cytokines in Sjögren's syndrome in combination with a better understanding of the effect of cytokines on exocrine dysfunction will aide the identification of the best therapeutic targets for Sjögren's syndrome.     

Clinical utilization of sialochemistry in Sjögren's syndrome

Sialochemistry was performed on the stimulated parotid secretion of a group of patients with Sjögren's syndrome (SS) having a Grade 4 iymphocytic infiltrate of their minor labial salivary glands and a normal control group. Parameters examined included flow rate, and concentration of sodium, potassium, chloride, urea, calcium, phosphate, total protein, IgA, IgG. albumin, amylase and lactoferrin. Although all SS patients had virtually no parotid secretion in the absence of stimulation, with a gustatory stimulation, 40% of the patients with SS had a relatively normal parotid flow rate, when compared with the control group. The SS patients, regardless of flow rate, exhibited a highly significant (p < 0.01) elevation in the concentration of sodium, chloride, IgA, IgG, and lactoferrin and a significant (p < 0.05) increase in albumin concentration, when compared with the control group. The phosphate level was significantly lower (p < 0.01) in SS patients than in the control group. The elevated IgA in SS was almost all 1 IS, in contrast to parotitis where 7S was a major contributor. In view of the variation in flow rate in SS patients chemical quantitation of selected salivary components can be a valuable aid in the differential diagnosis of this disease and in monitoring patients over time.     

Correlations between total protein,lysozyme, immunoglobulins,amylase, and albumin in stimulated whole saliva during daytime

The correlations between salivary proteins and the daytime variations are not known. The present study investigated the within-subject variation of correlations and concentrations between lysozyme, IgA, IgG, IgM, albumin, amylase, and total protein in stimulated whole saliva of healthy adults in the course of a 12-h period. After several practise sessions, unstimulated and stimulated whole saliva samples were collected five times daily (at 8 a.m., 11 a.m., 2 p.m., 5 p.m., and 8 p.m.) from 30 healthy university students. Flow rate and total protein concentration were used as covariates, and gender as a between-subject factor in the MANOVA analysis. After this adjustment, there was significant within-subject variation in salivary IgA (P &lt; 0.001), albumin (P &lt; 0.01), amylase (P &lt; 0.05), and total protein (P &lt; 0.001) concentrations. Total protein correlated significantly with amylase albumin and IgA through different samplings. In addition, IgG correlated with albumin and lysozyme in the course of 12 h. On the whole, the correlations between variables remained stable during repeated samplings. In addition, rankings of subjects for the variables tended to be maintained across different samplings (P &lt; 0.001). However, the observed within-subject variations in salivary IgA, albumin, amylase, and total protein concentrations suggest that these proteins are subject to short-term variation.     

Changes in whole saliva in patients with coeliac disease

Many systemic diseases impair salivary flow rate and composition and therefore incite oral pathological processes. This study analyses the composition of whole saliva in patients with diagnosed coeliac disease (CD) and in healthy controls, and monitors possible changes in saliva composition after a short oral gluten challenge. Paraffin-stimulated whole saliva was collected from 128 CD patients and 55 healthy controls. In a separate study, paraffin-stimulated whole saliva samples were collected from 33 CD patients and 10 controls both before and 24 h after an oral mucosal and submucosal gluten challenge. No difference in saliva flow rate was observed, but total protein (P相似文献   

Cytokine concentrations in stimulated whole saliva among patients with primary Sjögren's syndrome, secondary Sjögren's syndrome, and patients with primary Sjögren's syndrome receiving varying doses of interferon for symptomatic treatment of the condition: a preliminary study

Sj?gren's syndrome is an autoimmune disorder which causes diminished salivary flow due to autoimmune sialoadenitis. This decrease in saliva flow is the result of inflammation and atrophy of the salivary glands. Most treatment regimens are palliative in nature, but treatment with interferon (IFN) holds promise for Sj?gren's syndrome sufferers. Several studies have investigated cytokine concentrations in the salivary glandular tissues from Sj?gren's syndrome patients; however, there is little information concerning cytokine expression in saliva. This is especially true with respect to treatment modalities and their effects on local cytokines. A clinical study was conducted to determine salivary interleukin (IL)-6, IFN, and IL-2, concentrations among subjects diagnosed with primary and secondary Sj?gren's syndrome and a healthy control group. The primary Sj?gren's syndrome showed significantly higher salivary IL-2 and salivary IL-6 than the control and secondary Sj?gren's groups. There were no between group differences for salivary IFN concentrations. In addition, the study assessed salivary IL-6, IFN, and IL-2 concentrations among 18 Sj?gren's syndrome patients before and after administration of IFN via the oral mucosal route. The results of the study showed that the mean values for the pre- and post-treatment groups for stimulated whole saliva flow rates were 3.15 and 3.74 ml/5 min, respectively. The post-treatment group exhibited a 16.8% increase in stimulated whole saliva flow rates. The salivary IL-6 concentration was 53.3% lower for the post-treatment group (17.79) as compared to the baseline value (33.35). The values for salivary IFN and salivary total protein were virtually unchanged from their baseline values. Salivary IL-2 values, however, were 50% lower in the post-treatment group (3.07) when compared to their respective baseline values (6.10). The results of this study suggest that healthy individuals exhibit lower salivary IL-2 and IL-6 as compared to individuals suffering from primary and secondary Sj?gren's syndrome. The results also suggest that administration of IFN via the oral mucosal route may increase salivary flow rates and depress certain cytokines (IL-2, IL-6) associated with inflammatory destruction of salivary glandular tissues in Sj?gren's syndrome patients.     

Punch-biopsy of minor salivary glands in the diagnosis of Sjögren's syndrome

Abstract— The aim of the present investigation has been to examine whether the histopathologic changes in the minor salivary glands in patients with Sjögren's syndrome are of the same nature as the changes in the major salivary glands. Punch-biopsies were taken from the lower labial mucosa in 14 patients; 12 of these had a verified Sjögren's syndrome. In 10 patients changes in accordance with those of a benign lymphoepithelial lesion were present in the minor salivary glands. The technique was found valuable in the diagnosis of general disorders with salivary gland involvement.     

Salivary flow and composition in lymphoma patients before,during and after treatment with cytostatic drugs

To investigate the effect of modern, intensive chemotherapy on salivary flow rate and composition, 79 patients suffering from Hodgkin's disease or non-Hodgkin lymphoma were studied before, during and after administration of cytostatic drugs. 49 patients (mean age 49.9 years, 30 men, 19 women) completed the 1-year study. All patients who received radiotherapy or medication other than cytostatics were excluded. The results showed no marked differences in stimulated salivary flow rates, buffering capacities and amylase and total protein concentrations between the beginning and the end of the 12 month trial. However, significant increases in albumin secretion into saliva and salivary lysozyme concentrations were observed. Total salivary IgG, IgA and IgM concentrations decreased significantly during the cancer therapy but values returned to the baseline levels after termination of treatment. Despite the well-known cytolytic effect of anticancer drugs, chemotherapy need not therefore be permanently detrimental to saliva.     

The effect of parotid salivary flow rate on the levels of salivary antimicrobial proteins in patients with Sj?gren's syndrome.

OBJECTIVE: The purpose of this study was to examine the effect of salivary flow rate on the levels of antimicrobial salivary proteins in 24 patients with Sj?gren's syndrome and 22 age- and race-matched healthy control subjects. METHOD AND MATERIALS: Parameters examined included stimulated salivary flow rate, total salivary protein, lactoferrin, lysozyme, amylase, and secretory immunoglobulin A. RESULTS: The mean total salivary protein and the mean salivary amylase were significantly greater in patients than in controls. However, no significant difference was observed in the mean stimulated salivary flow rates or the levels of lactoferrin, lysozyme, or secretory immunoglobulin A of patients and controls. To examine the effect of salivary flow rate on the levels of salivary antimicrobial protein, the levels of these proteins in patients with salivary flow rate of < or = 0.3 mL/min per gland were compared to those in healthy controls with salivary flow rate > or = 0.4 mL/min per gland. Analyses showed the levels of lactoferrin to be significantly higher among patients than among controls. CONCLUSION: The levels of salivary amylase and lactoferrin may be influenced by the levels of salivary output in patients with Sj?gren's syndrome. The relationship between salivary flow rate and the levels of amylase and lactoferrin is not clear at the present time.     

The level of cariogenic microlorganisms in patients with Sjögren's syndrome

Sixteen patients with caries-inacthe Sjögren's syndrome with low parotid salivary flow rates (≤ 0.25 mL/min) and 18 caries-inactive control subjects with higher salivary flow rates were compared. Mutans streptococci (MS) and lactobacilll (LB) counts were measured by means of Dentocult® SM strip mutans and LB assays. The group with Sjögren's syndrome displayed higher counts of MS (P = 0.014) and LB (p = 0.003) when compared with controls. The results of this study indicate that patients with caries-inactive Sjögren's syndrome and low salivary flow may have higher colonization of cariogenic mlcro-organisms than healthy individuals.     

Salivary antibodies,amylase and protein from children with early childhood caries

The aim of this study was to analyze the organic composition of saliva from children without dental caries and children with early childhood caries (ECC). Two groups of 20 children varying in age from 12 to 47 months were selected: Group I, caries-free children; Group II, children with early childhood caries (ECC). Samples of saliva were collected from each subject and submitted to immunological and biochemical assays. Measurements of total salivary IgA, IgG and IgM were performed by using nephelometric techniques, while total protein concentrations and amylase activity were determined by colorimetric techniques. Comparisons of values between groups were made by using U Mann-Whitney test (p<0.05). Children with ECC presented significantly higher levels of total salivary IgA and IgG, while the mean values of amylase activity, total protein concentrations and total IgM were similar between the groups. In this study, the presence of ECC was associated with an increase in total salivary IgA.     

Protein, albumin and cystatin concentrations in saliva of healthy subjects and of patients with gingivitis periodonitis

Salivary protein, albumin and cystatin concentrations were investigated in subjects with a periodontium and in patients with gingivitis or periodontitis were signigicantly increased compared with healthy subjects. Salivary protein ans albumin appeared to be positively correlated in all the groups, which suggests that the increase in salivary protein concentration in subjects with gingivitis or perodontitis is caused by leakage of plasma proteins. Cystatin concentrations in saliva of subjects with periodontitis wer significantly increased when compared with the healthy group and the gingivitis group (p<0.01). In the gingivitis and perodontitis group, salivary custatin was only weakly correlated with albumin concentations, which suggests that the increased salivary cystatin activity found in subjects with gingivitis and periodontitis is derived from sources otehr than plasma.     

Salivary gland function and changes in patients with oral lichen planus

Abstract – Saliva analysis, sialography and histopathologic examination of labial salivary glands were performed on patients with oral lichen planus. Diseases connected with salivary gland function were also recorded. Saliva analysis regarding secrection rate, pH and buffer capacity in unstimulated and stimulated saliva was permormed on 39 patients. 87% of the patients exhibired a low or very low unstimulated secretion rate, the mean value being 0.14 ml/min. The rate of stimulated saliva, pH and buffer capicity did not deviate from normal reference values. Sialographic examination was performed on 18 patients, corresponding to 36 major salivary glands. Radiologic changes were seen in 89% patients. Histopathologic examination was performed on 15 patients. Lymphocytic infiltration, acinar atrophy, fibrosis, fatty degeneration or ducral changes were observed in the minor glands of all patients. Different degrees of acinar atrophy were present in 93% of the patients. Lymphorytic infiltration was seen in 12 patients (80%) of whom three exhibited focal accumulation as in Sjögren's syndrome. Since decreased salivary secretion and symptoms of joint diseases and keratoconjunctivitis sicca were frequently present, over a third of the patients showed clinical signs comparable to those of Sjögren's syndrome. A high frequency of gastrointestinal and endocrine diseases was also recorded, which suggests that a general exo and endocrine influence may be present in patients with oral lichen planus.     

Interindividual variation,correlations, and sex‐related differences in the salivary biochemistry of young healthy adults

A cross‐sectional observational study was conducted to evaluate interindividual biochemical variation in unstimulated whole saliva in a population of 268 systemically healthy young students, 18–30 yr of age, with no apparent caries lesions or periodontal disease. Salivary flow rate, protein content, pH, buffering capacity, mucins MUC5B and MUC7, albumin, secretory IgA, cystatin S, lactoferrin, chitinase, amylase, lysozyme, and proteases were measured using ELISAs and enzymatic activity assays. Significant differences were found between male and female subjects. Salivary pH, buffering capacity, protein content, MUC5B, secretory IgA, and chitinase activity were all lower in female subjects compared with male subjects, whereas MUC7 and lysozyme activity were higher in female subjects. There was no significant difference between sexes in salivary flow rate, albumin, cystatin S, amylase, and protease activity. Principal component analysis (PCA) and spectral clustering (SC) were used to assess intervariable relationships within the data set and to identify subgroups. Spectral clustering identified two clusters of participants, which were subsequently described. This study provides a comprehensive overview of the distribution and inter‐relations of a set of important salivary biochemical variables in a systemically healthy young adult population, free of apparent caries lesions and periodontal disease. It highlights significant gender differences in salivary biochemistry.     

Localization of lysozyme mRNA in the labial salivary glands by in situ hybridization in Sjögren's syndrome

Abstract – In this study, lysozyme mRNA in labial salivary glands has been localized with in situ hybridization technique using 35S-labeled hen lysozyme cDNA (cDNALZM) as a hybridization probe in normals and in patients with Sjögren's syndrome. 35S-DNALZM:mRNA hybrids were detected only in acinar serous cells, although lysozyme was identified in ductal cells using immunohistochemical techniques. Our results suggest that the serous acinar cells are the only site of lysozyme synthesis in small salivary glands. The presence of lysozyme in ductal cells may be a result of reabsorption from the saliva or concentration from the blood or surrounding tissues.     

Oral pilocarpine HCl stimulates labial (minor) salivary gland flow in patients with Sjögren's syndrome

Pilocarpine HCl has been shown to stimulate parotid and submandibular gland salivary flow. The purpose of this study was to determine whether this cholinergic-muscarinic drug also stimulates labial (minor) salivary gland (LSG) flow and to relate that with whole unstimulated salivary (WUS) flow rateS. Subjects diagnosed with primary Sjögren's syndrome (SS-1; n = 9) or secondary Sjögren's syndrome (SS-2; n = 9) were enrolled in this study after meeting stringent enrollment criteria. An age-gender matched control group was also enrolled. The labial saliva was collected in a standardized manner on Per-iopaper® for 5 min and the volume was analysed by the Periotron®.Whole unstimulated salivary samples were collected for 5 min by the method of Mandel and Wot-man (1976).Each subject was dosed with pilocarpine HCl (5 mg; tablets; p.o.).After 60 min the LSG flow as well as the WUS flow was determined again as previously. The results indicated a significant (>180%) increase in both labial salivary gland flow as well as whole salivary flow in the SS-1 and SS-2 subjects (mean ± S. e.m.): [SS-1: WUS = 0.1080 ± 0.03 vs 0.2242 ± 0.03 ml per 5 min; LSG = 93.1 ± 22.2 vs 167.8 ± 15.9 μl/5 min; P < 0.001; SS-2: WUS = 0.1384 ± 0.02 vs 0.2775 ± 0.09 ml per 5 min; LSG = 97.7 ± 20.2 vs 182.8 ± 17.9 μl per 5 min; P < 0.001]. These results indicate a significant increase in labial salivary gland flow as well as whole salivary flow as stimulated by pilocarpine HCI in Sjögren's syndrome patients.