Ilana Yaron

Elevated tear interleukin-6 levels in patients with Sjögren syndrome

OBJECTIVE The purpose of the study was to investigate interleukin-6 (IL-6) levels in the tear fluid and sera of patients with Sjögren syndrome (SS). PARTICIPANTS Twelve patients with primary SS and 12 normal control subjects participated. INTERVENTION Tear fluid and sera were obtained from the study and the control groups. Evaluation of tear fluid and sera IL-6 levels was done by using a quantitative enzyme-linked immunosorbent assay kit. All assays were carried out blindly with respect to diagnosis. MAIN OUTCOME MEASURES Tear fluid IL-6 levels were measured. RESULTS The mean concentration (+/- standard error) of IL-6 in the tears of patients with SS was elevated significantly compared to that of normal control subjects (88.6+/-16.2 vs. 42.1+/-10.6 pg/ml; P < 0.05). No significant differences were noted in the serum IL-6 levels between the two groups. A significant correlation (r = 0.742, P = 0.006) was found between tear fluid IL-6 levels and the focus score of lip biopsy specimens in patients with SS. CONCLUSION Tear fluid IL-6 levels may serve as an important marker for tear gland involvement in SS.

Hydroxychloroquine treatment for primary Sjögren’s syndrome: its effect on salivary and serum inflammatory markers

OBJECTIVE To evaluate the effect of hydroxychloroquine treatment on interleukin 6 (IL6), hyaluronic acid (HA), and soluble interleukin 2 receptor (sIL2R) concentrations in the saliva and serum of patients with primary Sjögren’s syndrome (SS). METHODS Fourteen SS patients treated with hydroxychloroquine 200 mg/day for 12 months were investigated in an open prospective study. Clinical parameters of efficacy and routine biochemical and haematological data to assess drug safety and tolerability were determined every three months. Salivary and serum IL6, sIL2R, and HA values were determined at study entry, 6 and 12 months, using ELISA and radiometric assays. RESULTS After hydroxychloroquine treatment, salivary IL6 concentrations decreased from 13.2 (1.2) to 7.3 (1.1) pg/ml (mean (SEM)) (p < 0.0001). Similarly, salivary HA concentrations were also reduced from 577.8 (120) to 200 (34) ng/ml (mean (SEM) (p < 0.003). Serum IL6 concentrations decreased from 5.4 (0.6) to 2.9 (0.2) pg/ml (mean (SEM) (p < 0.001), while serum HA concentrations remained unchanged. No change has been detected in salivary or serum sIL2R concentrations after 12 months of treatment with hydroxychloroquine. Treatment also resulted in significant reduction in erythrocyte sedimentation rate, serum γ globulin, and C reactive protein values while only partial clinical improvement was noted in some patients. A more pronounced decrease of salivary IL6 and HA levels was found in the two patients in whom a reduction in the swelling of the parotid gland was noted. CONCLUSION In this open label study of hydroxychloroquine treatment for SS a significant reduction of some salivary inflammatory markers was seen at the end of 12 months. Although during the treatment period only a partial clinical effect could be noted, the findings suggest that a double blind controlled study of hydroxychloroquine in SS is indicated.

Active leflunomide metabolite inhibits interleukin 1β, tumour necrosis factor α, nitric oxide, and metalloproteinase-3 production in activated human synovial tissue cultures

Background: Leflunomide is now an approved agent for the management of adult rheumatoid arthritis (RA). Its active metabolite A771726 inhibits de novo pyrimidine biosynthesis. Although considered to be an immunosuppressive agent, its mechanism of action remains obscure. Objectives: Evaluation of the leflunomide active metabolite A771726 (LEF) effect on interleukin 1β (IL1β), tumour necrosis factor (TNFα), nitric oxide (NO), and stromelysin (metalloproteinase-3 (MMP-3)) production by activated human synovial tissue in culture. Methods: Synovial tissue was obtained during surgery from patients undergoing total knee replacement owing to RA or osteoarthritis (OA), cut into small pieces, and cultured in Petri dishes with test materials as previously described. IL1β, TNFα, NO, and MMP-3 were measured in the culture media after 48 hours incubation with different doses of LEF by methods previously described. Results: LEF (0.3, 3, and 9 μg/ml) inhibited IL1β production in the presence of lipopolysaccharide (LPS; 3 μg/ml) in a dose dependent manner (p<0.01) at LEF 0.3 μg/ml. TNFα production in the presence of IL1β (1 ng/ml) was also inhibited in a dose dependent manner (p<0.05 at LEF 0.3 μg/ml). NO and MMP-3 production in the presence of LPS (3 μg/ml) was inhibited as well (p<0.01 at LEF 1 μg/ml and at LEF 0.3 μg/ml, respectively). Synovial cell viability evaluated by the tetrazolium salt XTT was unaffected by the LEF concentration used. There was no qualitative difference in the response of OA and RA synovial tissue. Conclusion: Leflunomide may modulate the rheumatoid articular process by inhibition of local production of IL1β, TNFα, NO, and MMP-3.

Saliva: An additional diagnostic tool in Sjögren's syndrome

OBJECTIVE To assess the available data and the place of salivary analysis in the diagnosis of Sjögrens syndrome (SS). METHODS A Medline search of English language articles published between 1985 and 1996 and a manual search of the reference lists of relevant articles formed the data sources. These were combined with our clinical and experimental experience in this field. Each method of salivary analysis was assessed according to study design, type of saliva used for the study, sensitivity/ specificity for the diagnosis of SS, and correlation to the histopathological findings. RESULTS Increased levels of salivary Na+, immunoglobulins (particularly IgA), anti-Ro and La antibodies, lactoferrin, lysozyme, beta 2 microglobulin, prostaglandin E2, thromboxane B2, interleukin-6, and hyaluronic acid have been detected in various studies. Results varied according to the different methods used for saliva collection. CONCLUSION Although many changes have been detected in various constituents of saliva in SS patients, no test has proved sensitive or specific enough for diagnosing SS.

Salivary eicosanoid concentration in patients with Sjögren's syndrome.

OBJECTIVE: To investigate eicosanoid concentrations in the saliva of patients with primary Sjögrens syndrome (SS). METHODS: Whole mixed saliva of 36 subjects was assayed for eicosanoid concentrations using a radioimmunoassay. Patients with primary SS having positive lip biopsy served as the study group; their results were compared with data from patients with dry mouth and negative lip biopsy (dry mouth group), and with a group of normal healthy controls. RESULTS: Concentrations of thromboxane B2 were significantly (p < 0.01) increased in 18 patients with primary SS compared with 10 patients with dry mouth and eight healthy normal controls (1.95 (SD 0.51) ng/ml saliva compared with 0.52 (0.1) ng/ml and 0.3 (0.1) ng/ml, respectively). Similarly, prostaglandin E2 concentrations were also significantly increased (p < 0.01) in 11 patients with primary SS compared with five patients with dry mouth and eight normal controls (3.75 (0.82) ng/ml saliva compared with 0.32 (0.1) ng/ml and 0.41 (0.1) ng/ml, respectively). CONCLUSION: Salivary concentrations of eicosanoids are significantly increased in patients with primary SS, and this may prove helpful in the diagnosis of this disease.

Salivary and serum hyaluronic acid concentrations in patients with Sjögren's syndrome

OBJECTIVE To evaluate salivary hyaluronic acid (HA) concentration in patients with primary Sjögren’s syndrome (SS). METHODS Salivary and serum HA concentrations were evaluated using a radiometric assay. Thirty nine patients with SS served as the study group and their results were compared with 19 patients having clinical symptoms and signs of dry mouth and with 10 normal controls. RESULTS Salivary HA concentrations were significantly increased (p < 0.05) in the 39 patients with SS compared with the 19 patients with dry mouth and the 10 normal controls (240.7 (38.5) v 99.8 (14.6) and 91.3 (7.9) ng/ml, respectively) (mean (SEM)). No significant differences were noted in the serum HA concentrations between the three groups (42 (3.9) v 36.3 (4.1) and 32 (4.3) ng/ml, respectively) (mean (SEM)). No correlation could be found between salivary HA concentrations and the focus score of lip biopsies, nor between salivary HA concentrations and erythrocyte sedimentation rate or other serological tests. CONCLUSION Increased salivary HA concentrations can serve as a marker of local inflammation and may be of value in the diagnosis of SS.

Acute induction of joint inflammation in the rat by poly I . poly C.

Intraarticular injection of interferon inducer, double-stranded polyinosinate-polycytidylate (Poly I · C) caused acute synovitis in rats. This acute inflammatory response was accompanied by an increased concentration of prostaglandin E (PGE) in the synovial tissue. Double-stranded polyadenylate-polyuridylate (Poly A · Poly U) was less potent than Poly I · Poly C in inducing synovitis and increasing PGE concentration, while single-stranded polyinosinate (Poly I) or polycytidylate (Poly C) were inactive in these respects. Intraarticular injection of partially purified mouse fibroblast interferon also induced synovial inflammation. The present results suggest that interferon may be a mediator of viral inflammatory responses.

Simvastatin Induces Apoptosis of Fibroblast-Like Synoviocytes

Background: Statins (3-Hydroxy-3-methylglutaryl-CoA reductase inhibitors) exert favorable effects on lipoprotein metabolism, but appeared to possess anti-inflammatory properties among others, as suggested by their ability to inhibit collagen-induced arthritis in mice. Their activity in fibroblast-like synovial cells (FLS) has not yet been studied. Objectives: To evaluate the effect of varying doses of simvastatin on apoptosis of FLS. Methods: Synovial tissue, obtained during total knee replacement due to osteoarthritis, was cut into small pieces and cultured in Petri dishes with test materials, as previously described. FLS were incubated for 48 hours with 1 μmol/ml, 5 μmol/ ml, 15 μmol/ml and 50 μmol/ml of simvastatin. Following incubation, apoptosis was analyzed by two-dimensional flow cytometry (FACS) using annexin V/PI staining according to the manufacturer’s instructions. Results: Different concentrations of simvastatin induced apoptosis of FLS. The level proportion of apoptotic cells of resting or activated with lipopolysaccharide (LPS; 3 μg/ml) FLS, not treated with simvastatin, was 21%. At 48 hours, the rate of apoptosis of activated fibroblasts, incubated with 1 μmol/ml, 5 μmol/ml, 15 and 50 μmol/ml was 22%, 32%, 48% and 41% respectively. Synovial cell viability evaluated by tetrazolium salt XXT was unaffected by the simvastatin concentration used. Conclusion: Varying concentrations of simvastatin induce apoptosis of activated fibroblast-like synoviocytes, suggesting another possible mechanism of anti-inflammatory effects of statins in inflammatory conditions.

Differential effects of 1,25-(OH)2D3 on acyl hydrolase and cyclooxygenase activities in Interleukin 1β-stimulated human synovial fibroblast cultures

Effects of proinflammatory cytokines on synovial fibroblasts are considered to play a role in the accumulation of prostaglandin E2 (PGE2) in the inflamed joint in rheumatoid arthritis. We have previously shown that interleukin 1 beta (IL-1 beta) induces PGE2 production in synovial fibroblast cultures and that this effect is suppressed by the active metabolite of vitamin D3, 1,25-(OH)2D3. To study the mechanism of the inhibitory action of 1,25-(OH)2D3, its effect on acyl hydrolase (arachidonic acid (AA) release) and cyclooxygenase (COX) activities were investigated. Our findings indicate that IL-1 beta caused an increase in AA release and COX activity and that in the presence of 1,25-(OH)2D3 AA release, but not COX activity, is suppressed. In comparison, dexamethasone, which also inhibits IL-1 beta induction of PGE2, had inhibitory effects on both parameters.