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.     

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.     

Cholinergic autoantibodies from primary Sjögren’s syndrome modulate submandibular gland Na+/K+‐ATPase activity via prostaglandin E2 and cyclic AMP

Passafaro D, Reina S, Sterin‐Borda L, Borda E. Cholinergic autoantibodies from primary Sjögren’s syndrome modulate submandibular gland Na ⁺ /K ⁺ ‐ATPase activity via prostaglandin E ₂ and cyclic AMP. Eur J Oral Sci 2010; 118: 131–138. © 2010 The Authors. Journal compilation © 2010 Eur J Oral Sci We demonstrate that patients with primary Sjögren’s syndrome (pSS) produce functional IgG autoantibodies that interact with the glandular M₃ muscarinic acetylcholine receptors (mAChRs). These autoantibodies act as a partial muscarinic agonist, increasing prostaglandin E₂ (PGE₂) and cyclic AMP production through modifying Na⁺/K⁺‐ATPase activity, but also interfere with the secretory effect of the parasympathetic neurotransmitter. The IgG from patients with pSS has two effects on the submandibular gland. On the one hand, it may act as an inducer of the proinflammatory molecule (PGE₂) that, in turn, inhibits Na⁺/K⁺‐ATPase activity. On the other hand, it plays a role in the pathogenesis of dry mouth, abolishing the Na⁺/K⁺‐ATPase inhibition and the net K⁺ efflux stimulation of the salivary gland in response to the authentic agonist pilocarpine, decreasing salivary fluid production.     

Microchimerism in labial salivary glands of hematopoietic stem cell transplanted patients

Oral Diseases (2011) 17 , 484–488 Objective: Microchimerism has been extensively investigated in autoimmune diseases, which display similarities with graft‐vs‐host disease. This study was conducted to investigate the presence of microchimerism in minor salivary glands of hematopoietic stem cell transplanted patients, one of the targets of graft‐vs‐host disease. Methods: Labial salivary glands biopsy specimens from 11 stem cell transplanted patients were analysed. The samples were grouped in control (five specimens from a female‐to‐female transplantation) and study group (five glands from male‐to‐female transplantation). One male transplanted patient was used as a positive control. Fluorescence in situ hybridization with Y‐chromosome probe and immunofluorescence with anticytokeratin AE1/AE3 and CD45 were used to identify Y‐chromosome positive glandular epithelial cells from allogeneic hematopoietic stem cell transplanted patients. Results: In the study group, all samples were positive to Y‐chromosome and cytokeratin AE1/AE3, in agreement with the pattern exhibited by male labial salivary gland. None of the samples from control group were positive to Y‐chromosome despite being positive to cytokeratin AE1/AE3. Positivity to CD45 was not relevant. Conclusion: Microchimerism in the labial salivary glands of sex‐mismatched stem cell transplanted patients is a real phenomenon. Further studies are necessary to elucidate the impact of this phenomenon on the clinical status of stem cell transplanted patients.     

Expression of CD163, interleukin‐10, and interferon‐gamma in oral squamous cell carcinoma: mutual relationships and prognostic implications

Tumor‐associated macrophages (TAMs) and their associated inflammatory cytokines represent the major inflammatory component of the stroma of many tumors and can affect prognosis in the case of neoplasms. The objective of this study was to determine the prognostic significance of CD163⁺ cells, interleukin‐10 (IL‐10), and interferon‐gamma (IFN‐γ) in oral lesions associated with oral squamous cell carcinoma (OSCC). The levels of CD163, IFN‐γ, and IL‐10 in the tissue samples of 240 patients with OSCC and 58 patients with other oral lesions were assessed by immunohistochemistry. Individuals with low IFN‐γ levels, high IL‐10 levels, and low CD163 levels were of special concern with respect to OSCC progression. We found that high levels of CD163, or a combination of low IFN‐γ levels, high IL‐10 levels, and low CD163 levels, were associated with poorer overall survival (OS). CD163⁺ cells provide better predictive power for OS in comparison with traditional markers, such as clinical stage and lymph node metastasis. Therefore, CD163⁺ cells may be effective prognostic predictors of OSCC. IL‐10 may also indicate poor outcomes when IFN‐γ secretion is low and the cells are CD163^?.     

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.     

Immunoprofile of focal lymphocytic infiltration in minor salivary glands of healthy individuals

Aim: To characterize the immunohistochemical profile of the inflammatory cells included in the focal lymphocytic infiltration in the minor salivary glands of healthy individuals. Materials and Methods: Tissue samples of the labial and palatal salivary glands from 46 postmortem subjects, demonstrating the presence of focal lymphocytic infiltration were quantitatively evaluated for the presence of T‐ and B‐cell lymphocytes, plasma cells and macrophages by immunohistochemical and morphometric methods. Results: B‐cell lymphocytes, the predominant cell population in labial (67.5%) and palatal salivary glands (60.8%), were more frequent than T‐cell lymphocytes in both glands (P < 0.001). Among the T‐cell lymphocytes, CD₄‐positive cells were significantly more prevalent than the CD₈‐positive cells (P < 0.001). Plasma cells were almost absent, comprising only 0.01% of the focal lymphocytic infiltration cells of the labial and palatal salivary glands. Conclusions: Focal lymphocytic infiltration in the samples of the salivary glands obtained from healthy individuals is devoid of plasma cells. This can serve as an additional means to differentiate between focal lymphocytic infiltration in patients with Sjögren's syndrome, in which plasma cells are abundant, and focal lymphocytic infiltration in individuals with other causes of focal sialadenitis.     

TACI-Fc gene therapy improves autoimmune sialadenitis but not salivary gland function in non-obese diabetic mice

Oral Diseases (2012) 18 , 365–374 Objective: Patients with Sjögren’s syndrome (SS) show aberrant expression of the B cell‐related mediators, B cell‐activating factor (BAFF), and a proliferation‐inducing ligand (APRIL) in serum and salivary glands (SGs). We studied the biological effect of neutralizing these cytokines by local gene transfer of the common receptor transmembrane activator and CAML interactor (TACI) in an animal model of SS. Material and Methods: A recombinant serotype 2 adeno‐associated virus (rAAV2) encoding TACI‐Fc was constructed, and its efficacy was tested in the SGs of non‐obese diabetic mice. Ten weeks later, SG inflammation was evaluated and serum and SG tissue were analyzed for inflammatory markers including immunoglobulins (Ig) and cytokines. Results: AAV2‐TACI‐Fc gene therapy significantly reduced the number of inflammatory foci in the SG, owing to a decrease in IgD⁺ cells and CD138⁺ cells. Moreover, IgG and IgM levels, but not IgA levels, were reduced in the SG. Overall expression of mainly proinflammatory cytokines tended to be lower in AAV2‐TACI‐Fc‐treated mice. Salivary flow was unaffected. Conclusion: Although local expression of soluble TACI‐Fc reduced inflammation and immunoglobulin levels in the SG, further research will have to prove whether dual blockade of APRIL and BAFF by TACI‐Fc can provide a satisfying treatment for the clinical symptoms of patients.     

Kinetics of Th17‐related cytokine expression in experimentally induced rat periapical lesions

Th17‐related cytokines are essential factors in various pathological states, including inflammatory bone destruction. This study investigated the contribution of Th17‐related cytokines to the progress of experimentally induced rat periapical lesions. Periapical pathoses were induced by unsealed exposure of the pulp chamber of the lower first molars. A variety of immunocompetent cells, including CD68⁺ macrophages, Ia antigen⁺ cells and TCRαβ⁺ T cells, were observed in the lesions. The expression levels of Th17‐related cytokines, IL‐17 and IL‐23, and of pro‐inflammatory cytokines, IL‐1β and IL‐6, were significantly increased at 14 days (expansion stage) compared with normal periapical tissues. The expression levels of Foxp3, a regulatory T cell (Treg)‐related gene, and of IL‐10, an anti‐inflammatory cytokine, were higher at 28 days (chronic stage) than at 14 days. These findings suggest that Th17‐related cytokines may be primary contributors to the initiation of periapical bone destruction, and that lesion expansion may be regulated by anti‐inflammatory mediators.     

Immunohistochemical assessment of CD1a‐positive Langerhans cells and their relationship with E‐cadherin in minor salivary gland tumors

J Oral Pathol Med (2012) 41 : 47–53 Objective: The aim of this study was to investigate the presence of CD1a‐positive Langerhans cells and their relationship with E‐cadherin in minor salivary gland tumors. Methods: Twenty‐seven minor salivary gland tumors were investigated using immunohistochemistry for CD1a and E‐cadherin. Results: A significant difference regarding the mean density of CD1a‐positive Langerhans cells was observed between pleomorphic adenomas and malignant tumors studied (P = 0.001). No CD1a‐positive cells were detected in most cases (n = 5) of cystic adenoid carcinomas. CD1a‐positive cells were detected in one mucoepidermoid carcinoma case, and six low‐grade polymorphous adenocarcinomas cases. Comparison of the mean density of CD1a‐positive cells between the three malignant tumors showed no significant difference (P = 0.127). No significant difference was observed in the presence of E‐cadherin between tumors (P = 0.73), but it was detected in 24 cases. Conclusions: The lack of CD1a‐positive in malignant salivary gland tumors facilitates the neoplastic development and suggests that these cells might be useful as auxiliary diagnostic and prognostic tool in minor salivary gland tumors. Furthermore, it is suggested that E‐cadherin mediates cell adhesion in these tumors although we did not demonstrate significance.     

Correlation of cytomegalovirus and human herpesvirus 7 with CD3+ and CD3+ CD4+ cells in chronic periodontitis patients

Thomasini RL, Bonon SH, Durante P, Costa SCB. Correlation of cytomegalovirus and human herpesvirus 7 with CD3 ⁺ and CD3 ⁺ CD4 ⁺ cells in chronic periodontitis patients. J Periodont Res 2012; 47: 114–120. © 2011 John Wiley & Sons A/S Background and Objective: Human chronic periodontitis is an inflammatory process characterized by dense accumulation of immune cells in the periodontal tissue. The periodontitis can lead to loss of teeth in the patient and the pathogenesis of this disease is not completely known. This study tested the hypothesis that chronic periodontitis‐affected sites can harbor betaherpesviruses and that viruses are linked to a profile of the inflammatory infiltrate. Material and Methods: Biopsies of periodontal tissue were taken from periodontitis‐affected patients and from healthy subjects. Immunohistochemistry was performed to count CD19⁺ B cells, CD3⁺ total T cells, T‐CD4⁺ and T‐CD8⁺ cell subsets, and PCR was performed to detect cytomegalovirus and human herpesvirus 6 and 7 in the samples. One slide of each sample was stained with Giemsa for histopathological examination and to evaluate the quality of the cellular infiltrate. Results: As expected, tissues collected from healthy subjects presented no significant level of inflammatory infiltration and were therefore excluded from immunostaining procedures. Results showed that CD19⁺ B cells were in higher number than CD3⁺ T cells in the periodontitis‐affected tissue, but this was not statistically significant. The T‐CD4⁺ lymphocyte subset was significantly higher than the T‐CD8⁺ lymphocyte subset (p = 0.004) in the samples. Cytomegalovirus and human herpesvirus 7 were found at periodontitis‐affected sites, but not in tissue collected from healthy subjects (p = 0.04 and p = 0.04, respectively). Human herpesvirus 6 was rarely detected. We found a correlation between cytomegalovirus and lower CD19⁺/CD3⁺ ratios (ratio < 0.9, p = 0.003) and between human herpesvirus 7 and lower CD19⁺/CD3⁺ ratios (ratio < 0.9, p = 0.003) and higher CD4⁺/CD8⁺ ratios (ratio > 1.1, p = 0.002). Conclusion: This study shows that cytomegalovirus and human herpesvirus 7 can be present at periodontitis‐affected sites but are uncommon at healthy periodontal sites. Moreover, our data suggest that cytomegalovirus can be related to an inflammatory infiltrate with predominance of CD3⁺ T cells, whereas human herpesvirus 7 can be associated with an infiltrate with predominance of T‐CD4⁺ cells. However, further studies are necessary to support this hypothesis. Herpesviruses could play a role in human chronic periodontitis by modulation of the T cell response.     

Wegener’s granulomatosis of the major salivary glands

J Oral Pathol Med (2012) 41 : 721–727 Wegener’s granulomatosis is one of the anti‐cytoplasmic autoantibodies (ANCA)‐associated vasculitides. Although it typically affects the lungs and kidneys, the head and neck are also involved in most cases but the site usually affected is the upper airway. However, there are 35 cases with well‐documented clinicopathological data in which Wegener’s granulomatosis manifested in the major salivary glands, most commonly the parotid. Twenty‐four patients presented with salivary signs and symptoms, in eight of whom there was no other presenting manifestation. These signs and symptoms may mimic infection or neoplasia and laboratory investigations, including ANCA serology and histopathology, may be non‐specific; thus, in 21 of the 35 patients (60%) there was a delay in diagnosis. Amongst the 21 were 11 of the 14 (78.6%) patients who presented with unilateral parotid disease and three of the five who died. Three other patients suffered permanent pulmonary, two renal and five facial nerve damage. This article reviews the literature on major salivary gland involvement in Wegener’s granulomatosis, which should be considered in the differential diagnosis of major salivary gland disease particularly if commoner causes have been excluded. A detailed medical history, and persistently inconclusive laboratory tests, could provide the clues that enable prompt diagnosis.     

Enumeration of T cell subsets with monoclonal antibodies in minor salivary glands of patients with rheumatoid arthritis

Abstract – Labial salivary glands of 51 patients with rheumatoid arthritis and those of 25 control patients were examined by the ANAE (acid α-naphthyl acetate esterase) technique to determine the percentages of B- and T-lymphocytes and mononuclear phagocytes (MPS cells). Using monoclonal antibodies (OKT3 for all T cells, OKT4 for helper/inducer T cells, OKT6 for thymocytes, and OKT8 for suppressor/cytotoxic T cells) T cell subsets were enumerated. B- lymphocytes predominated in both series of salivary glands, and the percentages of B and T cells were equal in both series. The absolute cell counts in the salivary glands of rheumatics were significantly higher (P<0.001) than in those of healthy controls. The number of OKT4⁺ cells was increased in rheumatics, leading to an elevated OKT4⁺/OKT8⁺ ratio when compared with that in controls (P<0.01). The results suggest that the basic phenomenon behind the B cell hyperactivity noticed in rheumatics might be due to increased activity of T helper cells rather than reduced number of T suppressor cells, which were shown to remain almost unaffected in the salivary glands of patients with rheumatoid arthritis.     

PLUNC protein expression in major salivary glands of HIV‐infected patients

Oral Diseases (2011) 17 , 258–264 Objective: To analyse and compare the expression of Palate, Lung, and Nasal Epithelium Clone (PLUNC) proteins in salivary glands from patients with and without AIDS (control group) using autopsy material. Methods: We analysed the expression of PLUNCs using immunohistochemistry in parotid (n = 45), submandibular (n = 47) and sublingual gland (n = 37) samples of AIDS patients [30 with normal histology, 21 with mycobacteriosis, 14 with cytomegalovirus (CMV) infection, 30 with chronic non‐specific sialadenitis, and 30 HIV‐negative controls. In situ hybridization (ISH) for SPLUNC 2 in the HIV‐negative group was performed. Results: SPLUNC 1 expression was detected in the mucous acini of submandibular and sublingual glands, and SPLUNC 2 were seen in the serous cells. LPLUNC 1 expression was only positive in the salivary ducts. There was a higher expression of SPLUNC 2 in AIDS patients with CMV infection and mycobacteriosis when compared with all other groups. The intensity of staining for SPLUNC 2 was greater around the lesions than the peripheral ones. ISH for SPLUNC 2 showed perinuclear positivity in the serous cells in all HIV‐negative cases. Conclusions: SPLUNC 1 and LPLUNC 1 proteins were similarly expressed in the salivary glands of AIDS patients and non‐HIV patients. CMV infection and mycobacteriosis increase SPLUNC 2 expression in serous cells in the salivary gland of AIDS patients.     

Innervation pattern and Ca2+ signalling in labial salivary glands of healthy individuals and patients with primary Sjögren's syndrome (pSS)

Abstract: We have characterised the innervation pattern and intracellular Ca²⁺-signalling in labial salivary glands (LSG) of 16 patients with primary Sjögren's syndrome (pSS) and 27 healthy controls. Numerous immunoreactive nerve fibers (IRF) containing vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase activating peptide (PACAP) were found around acini, ducts and blood vessels. Substance P (SP)-, neuropeptide Y-, tyrosine hydroxylase- and nitric oxide synthase-IRF were mainly surrounding ducts and blood vessels. The majority of pSS patients had inflamed LSG and the presence of focal lymphocytic infiltrates (FI) were more frequent and pronounced as compared with healthy controls. In areas with normal or diffusely inflamed LSG tissue, pSS patients demonstrated the same distribution of IRF as healthy controls with similar histology. However, IRF were absent in central areas of FI both in pSS and age-matched healthy controls. Although all pSS patients had hyposalivation, stimulation with acetylcholine, norepinephrine, phenylephrine, isoproterenol, VIP, PACAP, SP, adenosine 5'-triphosphate and uridine 5'-triphosphate induced the same increase in the intracellular free Ca²⁺ concentration in LSG acini from both pSS patients and healthy controls, indicating the presence of functional receptor systems in vitro .     

辅助性T细胞及其细胞因子与原发性合格伦综合征的相关性

舍格伦综合征(SS)是一种主要累及泪腺和唾液腺等外分泌腺的系统性自身免疫性疾病,分为原发性舍格伦综合征(pSS)和继发性舍格伦综合征(sSS).辅助性T(Th)细胞中Th1与Th2细胞分化不平衡,Th17细胞与Treg细胞免疫失衡可能对pSS的发生及其发展产生影响.Th细胞分泌的白细胞介素(IL)-6、IL-17、IL-21和IL-23等细胞因子在pSS发病过程中和介导SS小腺体内炎症的过程中可能起着重要的作用.其中,有关Th17细胞及其分泌的细胞因子的研究,可为pSS的治疗提供更为有力和关键的证据.     

Human T‐cell responses to oral streptococci in human PBMC‐NOD/SCID mice

We investigated cellular and humoral immune responses to oral biofilm bacteria, including Streptococcus mutans, Streptococcus anginosus, Streptococcus sobrinus, and Streptococcus sanguinis, in NOD/SCID mice immunized with human peripheral blood mononuclear cells (hu‐PBMC‐NOD/SCID mice) to explore the pathogenicity of each of those organisms in dental and oral inflammatory diseases. hu‐PBMC‐NOD/SCID mice were immunized by intraperitoneal injections with the whole cells of the streptococci once a week for 3 weeks. FACS analyses were used to determine the percentages of various hu‐T cell types, as well as intracellular cytokine production of interleukin‐4 and interferon‐γ. Serum IgG and IgM antibody levels in response to the streptococci were also determined by enzyme‐linked immunosorbent assay. S. anginosus induced a significant amount of the proinflammatory cytokine interferon‐γ in CD4⁺ and CD8⁺ T cells in comparison with the other streptococci. However, there was no significant differences between the streptococci in interleukin‐4 production by CD4⁺ and CD8⁺ T cells after inoculation. Further, S. mutans significantly induced human anti‐S. mutans IgG, IgG₁, IgG₂, and IgM antibodies in comparison with the other organisms. In conclusion, S. anginosus up‐regulated Th1 and Tc1 cells, and S. mutans led to increasing levels of their antibodies, which was associated with the induction of Th2 cells. These results may contribute to a better understanding of human lymphocyte interactions to biofilm bacteria, along with their impact on dental and mucosal inflammatory diseases, as well as endocarditis.     

Sialochemistry and cortisol levels in patients with Sjogren's syndrome

Oral Diseases (2012) 18 , 255–259 Objectives: (i) To determine whether salivary cortisol and electrolyte levels differ between patients with Sjogren’s syndrome (SjS) and healthy individuals. (ii) To assess correlations between whole‐saliva cortisol and some clinical manifestations in patients with SjS. Methods: A total of 24 healthy women (mean age 49.3 ± 9.8) served as controls (C) vis‐à‐vis 17 patients with SjS (mean age 55.5 ± 15.7). Salivary cortisol concentration was determined, and sialochemistry analysis was performed. Results: Significantly lower saliva flow rates and higher salivary chloride (Cl^?), potassium (K⁺), and Ca²⁺ levels were found in the SjS group. No significant differences or correlations were found in other parameters, including sodium (Na⁺), magnesium (Mg²⁺), phosphate (^?), urea (U), and salivary cortisol levels. Conclusion: Increased whole‐salivary output of Cl^? and K⁺ in SjS may reflect release from apoptotic rests of acinar cells after secondary necrosis. Normal levels of salivary Na⁺, Mg²⁺, and ^? argue against concentration effect, deranged tubular function or cortisol (mineralocorticosteroid) effect as the cause for these findings. Increased salivary Ca²⁺ levels probably reflect leakage of plasma Ca²⁺ through the injured oral mucosa in SjS. In spite of disease‐associated stress, salivary cortisol, a stress biomarker, was not increased, suggesting insufficient hypothalamus–pituitary–adrenal (HPA) axis response and/or local consumption of cortisol by lymphocyte infiltrates.