Ultrastructural study of Sj?gren's syndrome-like disease in MRL/l mice

Salivary glands of autoimmune MRL/l mice were examined ultrastructurally and by immunoelectron microscopy to further characterize the Sj?gren's syndrome-like disease in these animals. Major salivary glands from 12 female and 7 male MRL/l, two female MRL/n, and one female BALB/c mice were examined by electron microscopy and the glands from 4 female MRL/l mice were subjected to immunoelectron microscopy in order to detect Lyt-1 and Lyt-2 positive lymphoid cells. Mononuclear cell infiltrates were not seen in the salivary gland from the BALB mouse and occurred rarely in glands of MRL/n mice. However, in MRL/l mice, numerous lymphoid cells were present and acinar cells displayed low cytoplasmic density, cytoplasmic vacuolization and cellular lysis. Lymphoid cells were predominantly Lyt-1 positive although some Lyt-2 positive cells were observed. These results suggest that the MRL/l mouse represents a useful model for the study of the pathogenesis of Sj?gren's syndrome in man.     

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.     

Histochemical study of labial salivary glands in Sjögren's Syndrome

Abstract Human labial salivary gland biopsies of patients presenting connective tissue diseases associated with Sjögren's syndrome were submitted to a polysaccharide histochemistry study. The normal acinar secretion is an association of neutral polysaccharides with a sulphosialomucin. In Sjögren's syndrome, there is a great reduction in the secretory activity of the acinar cells, but no qualitative change was observed. The pathogenesis of this decreased production and its importance regarding the clinical manifestations of Sjögren's syndrome are discussed.     

Clinical and microbiological studies of periodontal disease in Sjögren's syndrome patients

Background: Little is known about the periodontal status of patients with Sjögren's Syndrome (SS), a chronic inflammatory autoimmune disease characterized by xerophthalmia and xerostomia. The aim of the present study was to evaluate whether the periodontal status of SS patients, in terms of clinical and microbiological parameters, differs from systemically healthy age‐ and gender‐matched controls. Methods: 8 primary SS and 10 secondary SS patients were examined in comparison with 11 control subjects. All patients were diagnosed by the European Community Criteria. Control subjects were systemically healthy and not undergoing periodontal treatment. The comparison of clinical status was made in terms of mean periodontal parameters (plaque index, gingival index, gingival recession, probing pocket depth, probing attachment level and bleeding on probing) as well as the frequency distribution of probing pocket depth and probing attachment level measurements. Microbiological assays of the subgingival dental plaque samples were carried out by both a chairside enzyme test (Periocheck^®) for the detection of peptidase activity (PA) and a polymerase chain reaction (PCR) analysis for 9 selected periodontal micro‐organisms (Actinobacillus actinomycetemcomitans, Fusobacterium nucleatum, Prevotella intermedia, Treponema denticola, Porphyromonas gingivalis, Eikenella corrodens, Campylobacter rectus, Bacteroides forsythus, Streptococcus oralis). Results: The occurrence, severity and extent of periodontal lesions were not significantly different between the 3 patient groups for all periodontal parameters examined. No significant differences in the sub‐gingival plaque samples from control, primary or secondary SS patients for the PA test, frequency or type of periodontal micro‐organisms observed. Conclusion: No significant differences could be detected in either clinical or microbiological parameters of primary or secondary SS patients compared with that of control subjects. The results of the present study thus support the notion that the periodontal status of patients with SS do not differ from systemically healthy age‐ and gender‐matched controls.     

Anti-salivary antibodies in primary Sjögren's syndrome

Earlier studies have described an antibody that recognized salivary ductal epithelium in sera from 15–50% of patients with primary Sjögren's syndrome; however, the specific salivary antigen in those studies was not identified. The present study further investigated this unknown salivary antigen. Twenty-nine of 31 patients (94%) with primary Sjögren's syndrome demonstrated IgG antinuclear antibodies that bound to an epithelial cell line with ductal characteristics derived from a human salivary gland. Seventy-seven percent of these patients had serum antibodies that bound to ductal cells of normal human parotid tissue after formalin fixation. Western blots of cell extracts, immunofluorescence, and adsorption studies indicated that SS-A/Ro and SS-B/La were the antigens recognized in the salivary cell line. The pattern of fluorescence seen when anti-SS-B/La bound to normal parotid tissue was identical to the fluorescence pattern of the anti-salivary ductal antibodies described in earlier literature.     

Reproducibility of biopsy grade in Sjögren's syndrome

Abstract: The purpose of this study was to examine the reproducibility of biopsy grades at various tissue depths in Sjögren’s syndrome. The biopsy grades of 38 minor salivary gland biopsies were examined at 6 μm, 50 μm, 100 μm, 150 μm, 200 μm, and 250 μm tissue depths. Tissue sections were stained with routine hematoxylin and eosin, graded I–IV, and compared with the initial “baseline” biopsy grade. The majority of the biopsies showed a wide range of grade variability at all depths. No tissue depth was consistently reproducible for any grade (P0.41, 0.64, 0.91, and 0.20, respectively). The difference between baseline grades and grades of deeper sections was sufficient to impact the diagnosis of Sjögren’s syndrome in approximately 60% of the biopsies (P<0.001). The overall result of this study suggests that examination of multiple sections of minor salivary gland biopsies is advisable to improve the reliability of the grade when evaluating 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.     

Management of dry mouth in Sjögren's syndrome

The management of dry mouth is essential for patients with Sjögren's syndrome. The symptomatic treatment has included using air humidifiers, rinsing the mouth with water or mouthwash, the application of a salivary substitute and administration of secretagogues. There are three secretagogues suitable for the alleviation of dry mouth in Sjögren's syndrome patients in Japan; cevimeline hydrochloride hydrate (cevimeline), pilocarpine hydrochloride, and anetholtrithione. A relationship between the effect of cevimeline on saliva secretion and the degree of salivary gland destruction evaluated by sialography and histopathological findings in the labial minor salivary glands has been reported. These diagnostic approaches could provide useful prognostic information on the efficacy of cevimeline in Sjögren's syndrome patients. Concomitantly, a bite guard was suggested as an effective lubricating device because it maintains the lubricants in the proper location. In addition, the management of the complications of dry mouth, such as tooth caries, periodontitis and oral candidiasis, which all lead to a reduction in the QOL, is also important. Both the prevention and treatment of erythematous candidiasis is especially important in the management of Sjögren's syndrome.     

An association between salivary gland disease and serological abnormalities in Sjögren's syndrome

In the labial salivary glands (LSGs) of 16 primary and 18 secondary Sjögren's syndrome (SS) patients, infiltrating lymphocytes were histologically and immunohistochemically examined: also, the serum levels of rheumatoid factor, antinuclear antibodies, anti-DNA antibodies, anti-SS-A and anti-SS-B antibodies, and immunoglobulins (including IgG, IgM and IgA) were all assayed. An immunohistochemical analysis of the lymphocyte subsets in LSGs revealed that severe lymphocytic infiltration was frequently accompanied by marked B cell accumulation both in primary and secondary SS patients. Furthermore, local B cell accumulation was also closely associated with elevated levels of anti-SS-A and anti-SS-B antibodies and IgG, and this association was statistically significant in the group with primary SS but not in the group with secondary SS. Thus, local lymphocytic infiltration, especially B cell accumulation, in the salivary glands is suggested to be involved in serological abnormalities in primary SS. while complicated autoimmune diseases other than SS may also be involved in serological abnormalities in secondary SS.     

The minor salivary gland proteome in Sjögren's syndrome

Objective:  To identify the global protein expression (the proteome) in the minor salivary glands from primary Sjögren's syndrome (pSS) patients and non-SS controls.
Materials and methods:  Minor labial salivary glands were obtained from six pSS patients and from six age-matched non-SS controls, lysed in SDS buffer and pooled into two groups, respectively. The lysates were analysed by liquid chromatography electrospray ionization combined with tandem mass spectrometry. Also, the proteins were separated by two-dimensional polyacrylamide gel electrophoresis and protein spots were subjected to mass spectrometry.
Results:  Heat shock proteins, mucins, carbonic anhydrases, enolase, vimentin and cyclophilin B were among the proteins identified. The differences in the proteomes of minor salivary glands from pSS patients and non-SS controls were mainly related to ribosomal proteins, immunity and stress. Alpha-defensin-1 and calmodulin were among six proteins exclusively identified in pSS patients.
Conclusion:  We have identified several minor salivary gland proteins that may have implications for clarifying the SS pathophysiology. This experiment adds to the knowledge of proteins produced in salivary glands in health and disease, and may form the basis of further studies on biomarkers of prognostic and diagnostic value.     

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.     

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.     

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.     

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.     

Frequency and predictive value of the clinical manifestations in Sjögren's syndrome

The purpose of this study was to examine the frequency and predictive value of glandular and extraglandular manifestations in S ogren's syndrome (SS). The clinical profiles of 169 SS patients were compared to those of 44 non-SS controls. The specific symptoms examined were oral, ocular, vaginal, gastric, pulmonary, skin, joint and muscle pain. Statistical analyses were performed on both individual and grouped symptoms. Chi-squared analyses showed that the frequency of all symptoms was significantly higher among patients than controls. Stepwise discriminant analysis of individual symptoms suggests that the combined symptoms of dry mouth, sore mouth, and dry eyes correctly classified 93% of SS and 97.7% of the controls. While grouped gastric, muscle, psychological, vaginal, skin, nasal, and thyroid symptoms correctly classified 64.3% of SS and 86.1% of the controls. This is the first study to examine the diagnostic value of multi-system manifestation in SS. The overall results suggest that a comprehensive questionnaire of various symptoms may assist the diagnosis of SS. The high predictive value of the combined symptoms confirms their value in the evaluation of SS.     

A histopathological study of lymphoepithelial island formation in labial salivary glands in patients with primary Sjögren's syndrome

Abstract: The proliferative status of lymphoepithelial islands in the labial salivary glands of primary Sjögren’s syndrome (pSS) patients was investigated by counting the number of argyrophilic nucleolar organizer regions (AgNORs) in epithelial cells constituting the islands. The islands were classified into four groups and evaluated in terms of total area and three discrete zones of the islands. In each pSS group, the mean AgNOR number per total island epithelial cell nucleus was significantly higher than in control ductal epithelial cells. The zonal AgNOR number fluctuated during the process of island formation but became more uniform as the islands developed. Furthermore, statistically significant trends among the four pSS groups were observed in the ratio of T lymphocytes, B lymphocytes and plasma cells surrounding the islands. The results indicated that the islands are highly proliferative once island formation begins and that zonal island cell proliferation may be associated with the inflammatory cells.     

Primary Sjögren's syndrome: salivary gland function and clinical oral findings

OBJECTIVE: To evaluate salivary gland function, saliva composition and oral findings in patients with primary Sjogren's syndrome (pSS) subdivided into patients with and without focus score ≤1 (FS) and/or antibodies to SSA/SSB (AB) as well as in healthy controls. SUBJECTS AND METHODS: Unstimulated (UWS) and chewing stimulated (SWS) whole saliva, and stimulated parotid saliva (SPS) were collected in 16 patients fulfilling the European classification criteria for pSS subdivided into those with FS and/or AB (n= 8) and those without FS and AB (n= 8), and in age-matched (n= 14) and young healthy controls (n= 13).UWS and SWS were analysed for Na⁺ and K⁺.SPS was analysed for Na⁺, K⁺, statherin, and proline-rich proteins (PRPs).Sicca symptoms, DMFT/DMFS, plaque (PI) and gingival (GI) scores, periodontal pocket depth (PPD), and mucosal status were recorded. RESULTS: The young healthy controls had lower UWS as compared to the aged controls (P= 0.03).However, the aged controls had higher DMFT/DMFS (P < 0.001) and PI, GI and PPD (P < 0.01).Patients with FS and/or AB generally had lower saliva secretory rates than patients without FS and/or AB (P= 0.01 for UWS and SPS) and age-matched healthy controls (P= 0.001). There was no significant difference in the content of Na⁺ and K⁺, statherin and PRPs between groupS. Patients with FS and/or AB had the highest frequency of oral mucosal changes and higher DMFT/DMFS than patients without FS and/or AB and healthy controls (P < 0.01).However, PI, GI, and PPD did not differ significantly. CONCLUSION: Patients with FS and/or AB had lower salivary secretory rates, higher DMFT/DMFS, and more oral mucosal changes than patients without FS and/or AB.Additionally, data suggest that salivary gland function in healthy individuals do not decrease with age.