ABSTRACT
OBJECTIVES: The aims of the study were to investigate the prevalence of impaired sensation after minor salivary gland biopsy (MSGB) in two Swedish centres [Karolinska University Hospital (KUH) and Skåne University Hospital (SUH)] and to assess its impact on quality of life (QoL) and associated risk factors. METHOD: A questionnaire including questions regarding the presence of impaired sensation, impact on QoL, and impact on everyday life was sent to patients who had undergone MSGB between 2007 and 2016, and their medical notes were scrutinized. RESULTS: The study included 630 patients (505 from KUH and 125 from SUH). In KUH the biopsies were performed by rheumatologists and in SUH by dentists or oral and maxillofacial surgeons (OMSs). Long-standing, probably permanent, impaired sensation after MSGB was reported by 21% of patients, and was associated with lower age and absence of anti-SSA antibodies. Patients with long-standing impaired sensation reported the inconvenience (1-10) of impaired sensation as 4.0 (2.0-7.0) [median (interquartile range)], and 32% reported an influence on their QoL, the reported influence (1-10) on everyday life being 3.0 (1.0-5.0). When comparing the outcomes from KUH and SUH, patients from SUH reported a significantly lower frequency of long-standing impaired sensation (14% vs 23%; p = 0.02). CONCLUSION: A high frequency of long-standing impaired sensation after MSGB was found among patients who had undergone MSGB, although it had a low impact on everyday life. The complication frequency was less pronounced when a dentist or an OMS had performed the biopsy.
Subject(s)
Salivary Glands, Minor , Sjogren's Syndrome , Humans , Salivary Glands, Minor/pathology , Sjogren's Syndrome/pathology , Retrospective Studies , Quality of Life , Sweden/epidemiology , Hypesthesia/pathology , Biopsy/adverse effectsABSTRACT
BACKGROUND: To assess the risk of incident cardiovascular disease in patients with primary Sjögren's syndrome, overall and stratified by Ro/SSA and La/SSB autoantibody status. METHODS: A cohort of patients with primary Sjögren's syndrome in Sweden (n = 960) and matched controls from the general population (n = 9035) were included, and data extracted from the National Patient Register to identify events of myocardial infarction, cerebral infarction and venous thromboembolism. Hazard ratios were estimated using cox proportional hazard regressions. RESULTS: During a median follow-up of 9.5 years, the overall hazard ratio (HR) was 1.6 (95% CI 1.2-2.1) for myocardial infarction, 1.2 (95% CI 0.9-1.7) for cerebral infarction and 2.1 (95% CI 1.6-2.9) for venous thromboembolism. Patients positive for both Ro/SSA and La/SSB autoantibodies had a substantially higher risk of cerebral infarction (HR 1.7, 95% CI 1.0-2.9) and venous thromboembolism (HR 3.1, 95% CI 1.9-4.8) than the general population. These risks were not significantly increased in Ro/SSA- and La/SSB-negative patients. Among autoantibody-positive patients, the highest HR of cerebral infarction was seen after ≥10 years disease duration (HR 2.8, 95% CI 1.4-5.4), while the HR for venous thromboembolism was highest 0-5 years after disease diagnosis (HR 4.7, 95% CI 2.3-9.3) and remained high throughout disease duration. CONCLUSIONS: Primary Sjögren's syndrome is associated with a markedly increased risk of cardiovascular disease and the presence of Ro/SSA and La/SSB autoantibodies identify the subgroup of patients carrying the highest risk. These findings suggest that monitoring and prevention of cardiovascular disease in this patient group should be considered.
Subject(s)
Antibodies, Antinuclear/blood , Cerebral Infarction/etiology , Myocardial Infarction/etiology , Sjogren's Syndrome/complications , Venous Thromboembolism/etiology , Biomarkers/blood , Case-Control Studies , Cerebral Infarction/immunology , Female , Humans , Male , Middle Aged , Myocardial Infarction/immunology , Risk Factors , Sjogren's Syndrome/immunology , Sweden , Venous Thromboembolism/immunologyABSTRACT
OBJECTIVE: Environmental factors have been suggested in the pathogenesis of rheumatic diseases. We here investigated whether infections increase the risk of developing primary Sjögren's syndrome (pSS). METHODS: Patients with pSS in Sweden (n = 945) and matched controls from the general population (n = 9048) were included, and data extracted from the National Patient Register to identify infections occurring before pSS diagnosis during a mean observational time of 16.0 years. Data were analysed using conditional logistic regression models. Sensitivity analyses were performed by varying exposure definition and adjusting for previous health care consumption. RESULTS: A history of infection associated with an increased risk of pSS (OR 1.9, 95% CI 1.6-2.3). Infections were more prominently associated with the development of SSA/SSB autoantibody-positive pSS (OR 2.7, 95% CI 2.0-3.5). When stratifying the analysis by organ system infected, respiratory infections increased the risk of developing pSS, both in patients with (OR 2.9, 95% CI 1.8-4.7) and without autoantibodies (OR 2.1, 95% CI 1.1-3.8), whilst skin and urogenital infections only significantly associated with the development of autoantibody-positive pSS (OR 3.2, 95% CI 1.8-5.5 and OR 2.7, 95% CI 1.7-4.2). Furthermore, a dose-response relationship was observed for infections and a risk to develop pSS with Ro/SSA and La/SSB antibodies. Gastrointestinal infections were not significantly associated with a risk of pSS. CONCLUSIONS: Infections increase the risk of developing pSS, most prominently SSA/SSB autoantibody-positive disease, suggesting that microbial triggers of immunity may partake in the pathogenetic process of pSS.
Subject(s)
Infections/complications , Sjogren's Syndrome/etiology , Adult , Aged , Female , Humans , Male , Middle Aged , Prospective Studies , Risk Assessment , Sjogren's Syndrome/epidemiologyABSTRACT
B cells play a key role in the pathogenesis of primary Sjögren's syndrome (pSS). The aim of this study was to analyse the transcriptome of CD19+ B cells from patients with pSS and healthy controls to decipher the B cell-specific contribution to pSS. RNA from purified CD19+ B cells from 12 anti-SSA antibody-positive untreated female patients with pSS and 20 healthy blood donors was subjected to whole transcriptome sequencing. A false discovery rate corrected significance threshold of α < 0.05 was applied to define differential gene expression. As validation, gene expression in B cells from 17 patients with pSS and 16 healthy controls was analysed using a targeted gene panel. RNA-sequencing identified 4047 differentially expressed autosomal genes in pSS B cells. Upregulated expression of type I and type II interferon (IFN)-induced genes was observed, establishing an IFN signature in pSS B cells. Among the top upregulated and validated genes were CX3CR1, encoding the fractalkine receptor involved in regulation of B-cell malignancies, CCL5/RANTES and CCR1. Increased expression of several members of the TNF superfamily was also identified; TNFSF4/Ox40L, TNFSF10/TRAIL, TNFSF13B/BAFF, TNFRSF17/BCMA as well as S100A8 and -A9/calprotectin, TLR7, STAT1 and STAT2. Among genes with downregulated expression in pSS B cells were SOCS1 and SOCS3, CD70 and TNFAIP3/A20. We conclude that B cells from patients with anti-SSA antibody-positive pSS display immune activation with upregulated expression of chemokines, chemokine receptors and a prominent type I and type II IFN signature, while suppressors of cytokine signalling are downregulated. This adds insight into the autoimmune process and suggests potential targets for future functional studies.
Subject(s)
B-Lymphocytes/immunology , CX3C Chemokine Receptor 1/metabolism , Interferon Type I/immunology , Interferon-gamma/immunology , OX40 Ligand/metabolism , Sjogren's Syndrome/immunology , Adult , Aged , Antigens, CD19/metabolism , Autoantibodies/immunology , Autoantigens/immunology , Chemokine CCL5/metabolism , Female , Gene Expression Profiling , Gene Expression Regulation/immunology , Humans , Middle Aged , RNA, Small Cytoplasmic/immunology , Receptors, CCR1/metabolism , Ribonucleoproteins/immunology , Signal Transduction/immunology , Transcriptional Activation/immunology , Transcriptome/geneticsABSTRACT
Genetic investigations of Sjögren's syndrome (SS) have identified a susceptibility locus at p23.3 of chromosome 11, which contains the CXCR5 gene. C-X-C motif chemokine receptor 5 (CXCR5) is a chemokine receptor expressed on B and T cell subsets, and binds the chemotactic ligand C-X-C motif chemokine ligand 13 (CXCL13). In this study we aimed to link the genetic association with functional effects and explore the CXCR5/CXCL13 axis in SS. Expression quantitative trait loci analysis of the 11q23.3 locus was performed using B cell mRNA expression data from genotyped individuals. Lymphocyte surface markers were assessed by flow cytometry, and CXCL13 levels by a proximity extension assay. CXCR5+ and CXCL13+ cells in minor salivary glands were detected using immunohistochemistry. Our results demonstrated that SS-associated genetic polymorphisms affected the expression of CXCR5 (P < 0·01). Notably, a decreased percentage of CXCR5+ cells, with lower CXCR5 expression, was observed for most circulating B and T cell subsets in SS patients, reaching statistical significance in CD19+ CD27+ immunoglobulin (Ig)D+ marginal zone (P < 0·001), CD19+ CD27+ IgD- memory (P < 0·05) and CD27-IgD double-negative (P < 0·01) B cells and CD4+ CXCR3- CCR6+ Th17 cells (P < 0·05). CXCL13 levels were increased in patient plasma (P < 0·001), and immunohistochemical staining revealed expression of CXCL13 and higher numbers of CXCR5+ cells (P < 0·0001) within focal infiltrates and interstitially in salivary glands of SS patients. In conclusion, we link a genetic susceptibility allele for SS to a functional phenotype in terms of decreased CXCR5 expression. The decrease of CXCR5+ cells in circulation was also related to homing of B and T cells to the autoimmune target organ. Therapeutic drugs targeting the CXCR5/CXCL13 axis may be useful in SS.
Subject(s)
B-Lymphocyte Subsets/immunology , Chemokine CXCL13/blood , Receptors, CXCR5/blood , Sjogren's Syndrome/blood , T-Lymphocyte Subsets/immunology , Adolescent , Adult , Chemokine CXCL13/metabolism , Chromosomes, Human, Pair 11/genetics , Female , Genetic Predisposition to Disease/genetics , Humans , Inflammation/immunology , Male , Middle Aged , Polymorphism, Genetic/genetics , Receptors, CXCR5/biosynthesis , Sjogren's Syndrome/genetics , Sjogren's Syndrome/immunology , Young AdultABSTRACT
OBJECTIVES: To define the incidence rate of primary Sjögren's syndrome (pSS) and the prevalence of extraglandular manifestations (EGMs) at the time of diagnosis of pSS in a prospective, population-based manner. METHOD: This study included all consecutive patients referred to the Department of Rheumatology at Karolinska University Hospital for the investigation of incident pSS from 1 January 2007 to 31 December 2011. Investigation was according to the current criteria for pSS, and examination with a focus on the presence of EGMs was performed. RESULTS: Of the referred individuals, 199 out of 781 were diagnosed with pSS. We found an annual incidence rate of pSS in the Karolinska University Hospital catchment area of 3.1 [95% confidence interval (CI) 2.3-4.3] cases per 100 000 adult inhabitants. The female/male ratio of incident cases was 14/1 [frequency (female) = 0.93, 95% CI 0.89-0.96]. In our cohort, we noted lower figures for severe EGMs such as lung and neurological involvement than previously reported for prevalent pSS. The frequency of autoantibodies including antinuclear antibodies (ANA), anti-Ro/SSA, and anti-La/SSB was also lower compared to other cohorts. In our study, autoantibody-positive patients had cytopaenia significantly more often, and in patients older than 60 years primary biliary cirrhosis (PBC) was more common. CONCLUSIONS: The incidence rate of pSS is 3.1 (95% CI 2.3-4.3) per 100 000 person-years. The prevalence of autoantibodies may be lower than previously reported, and at diagnosis, patients with pSS have few severe EGMs.
Subject(s)
Lung Diseases/epidemiology , Nervous System Diseases/epidemiology , Sjogren's Syndrome/epidemiology , Sjogren's Syndrome/immunology , Adult , Aged , Autoantibodies/blood , Biopsy , Humans , Incidence , Longitudinal Studies , Lung Diseases/etiology , Male , Middle Aged , Nervous System Diseases/etiology , Prevalence , Prospective Studies , Retrospective Studies , Salivary Glands/pathology , Sjogren's Syndrome/complications , Sweden/epidemiologyABSTRACT
Ro52 is an E3 ubiquitin ligase with a prominent regulatory role in inflammation. The protein is a common target of circulating autoantibodies in rheumatic autoimmune diseases, particularly Sjögren's syndrome (SS). In this study we aimed to investigate the expression of the SS target autoantigen Ro52 in salivary glands of patients with primary Sjögren's syndrome (pSS). Ro52 expression was assessed by immunohistochemical staining of paraffin-embedded and frozen salivary gland biopsies from 28 pSS patients and 19 non-pSS controls from Swedish and Norwegian registries, using anti-human Ro52 monoclonal antibodies. The degree and pattern of staining and inflammation was then evaluated. Furthermore, secreted Ro52 protein was measured in saliva and serum samples from the same individuals through a catch-enzyme-linked immunosorbent assay (ELISA). Ro52 was highly expressed in all the focal infiltrates in pSS patients. Interestingly, a significantly higher degree of Ro52 expression in ductal epithelium was observed in the patients compared to the non-pSS controls (P < 0·03). Moreover, the degree of ductal epithelial expression of Ro52 correlated with the level of inflammation (Spearman's r = 0·48, P < 0·0120). However, no secreted Ro52 protein could be detected in serum and saliva samples of these subjects. Ro52 expression in ductal epithelium coincides with degree of inflammation and is up-regulated in pSS patients. High expression of Ro52 might result in the breakage of tolerance and generation of Ro52 autoantibodies in genetically susceptible individuals. We conclude that the up-regulation of Ro52 in ductal epithelium might be a triggering factor for disease progression in SS.
Subject(s)
Ribonucleoproteins/metabolism , Saliva/metabolism , Salivary Duct Calculi/metabolism , Salivary Glands/pathology , Sjogren's Syndrome/diagnosis , Adult , Aged , Biopsy , Cohort Studies , Female , Humans , Immune Tolerance , Immunohistochemistry , Inflammation/immunology , Male , Middle Aged , Ribonucleoproteins/immunology , Salivary Duct Calculi/immunology , Sjogren's Syndrome/immunology , Up-Regulation , Young AdultABSTRACT
The genetic background of primary Sjögren's syndrome (pSS) is partly shared with systemic lupus erythematosus (SLE). Immunoglobulin G Fc receptors are important for clearance of immune complexes. Fcγ receptor variants and gene deletion have been found to confer SLE risk. In this study, four Fcγ receptor single-nucleotide polymorphisms (SNPs) and one copy number variation (CNV) were studied. Swedish and Norwegian pSS patients (N=527) and controls (N=528) were genotyped for the Fcγ receptor gene variant FCGR2A H131R (rs1801274) by the Illumina GoldenGate assay. FCGR3A F158V (rs396991) was analysed in 488 patients and 485 controls, FCGR3B rs447536 was analysed in 471 patients and 467 controls, and FCGR3B rs448740 was analysed in 478 cases and 455 controls, using TaqMan SNP genotyping assays. FCGR3B CNV was analysed in 124 patients and 139 controls using a TaqMan copy number assay. None of the SNPs showed any association with pSS. Also, no FCGR3B CNV association was detected. The lack of association of pSS with Fcγ receptor gene variants indicates that defective immune complex clearance may not be as important in pSS pathogenesis as in SLE, and may point to important differences between SLE and pSS.
Subject(s)
Receptors, IgG/genetics , Sjogren's Syndrome/genetics , Adult , Aged , Case-Control Studies , Female , Gene Deletion , Genetic Association Studies , Humans , Male , Middle Aged , Norway , Polymorphism, Single Nucleotide , SwedenABSTRACT
The Ro52 protein of the Ro/SSA antigen was recently defined as an E3 ligase controlling cytokine production. Autoantibodies from systemic lupus erythematosus (SLE) patients targeting the Ro52-RING domain, containing the E3 ligase activity, have been shown to inhibit the E3 ligase activity of Ro52. The objective of the present study was to investigate correlations between clinical parameters in patients with SLE and levels of Ro/SSA (Ro52 and Ro60) and La/SSB autoantibodies, including autoantibodies directed towards the functional RING and B-box domains of the Ro52 protein. SLE patients (n=232) were clinically examined and disease activity indices collected concurrently to blood sampling. The samples were analyzed for immunological parameters including autoantibodies. Ro52 autoantibody levels were associated with more variables than the other analyzed antibodies and were significantly associated with several individual items related to sSS and the diagnosis of sSS itself (p=0.004). Other associated variables were high sedimentation rate (p=0.0003), levels of immunoglobulins (p=0.0003), and an inverse correlation with levels of lymphocytes (p=0.003) and leukocytes (p=0.01). Antibodies to the RING domain of Ro52, which is the functionally active domain with E3 ligase activity, were significantly correlated with disease activity as measured by the SLAM score. We conclude that autoantibodies against Ro52 and in particular its functional RING domain are important in lupus patients and associated with several clinical and laboratory features of the disease. The impact on disease activity of Ro52-RING specific antibodies was especially noted, and could imply a functional role for these autoantibodies in inhibiting Ro52 activity, which is important for the control of proinflammatory cytokine production, including type 1 interferons.
Subject(s)
Autoantibodies/physiology , Autoantigens/immunology , Lupus Erythematosus, Systemic/immunology , Ribonucleoproteins/immunology , Adult , Autoantibodies/biosynthesis , Female , Humans , Immunoglobulin G/biosynthesis , Immunoglobulin G/physiology , Lupus Erythematosus, Systemic/physiopathology , Male , Middle Aged , SS-B AntigenABSTRACT
BACKGROUND: Recent studies in experimental models and in vitro indicate lowering of IL-17/Th17 as an important mechanism of interferon-beta (IFN-ß) treatment in multiple sclerosis (MS). MATERIAL AND METHODS: In this longitudinal study of MS patients (n=25), spontaneous and myelin antigen-induced secretion of IL-4, IFN-γ and IL-10 (ELISPOT), mitogen stimulated secretion of IL-13 and IL-17A (ELISA) and circulating cytokine levels (Luminex) were recorded at inclusion and after 1.5, 3, 6 and 12months of IFN-ß treatment. RESULTS: Early changes were noted for IL-4, while after one year of treatment the only recorded significant effects were a decrease in secreted IL-17A levels and an increase in IL-10 secreting cells. While IL-17A levels tended to be higher in non-responders (n=8), the decrease in IL-17A levels seemed to be more pronounced in responders (n=17) showing significantly lower IL-17A levels after one year as compared with non-responders. CONCLUSION: IFN-ß treatment seems to mainly affect IL-17/IL-10-associated pathways rather than the IFN-γ/IL-4 axis.
Subject(s)
Interferon-beta/administration & dosage , Interferon-gamma/physiology , Interleukin-10/physiology , Interleukin-13/physiology , Interleukin-17/physiology , Interleukin-4/physiology , Multiple Sclerosis/drug therapy , Adult , Aged , Biomarkers/blood , Cohort Studies , Drug Administration Schedule , Female , Follow-Up Studies , Humans , Longitudinal Studies , Male , Middle Aged , Multiple Sclerosis/blood , Treatment OutcomeABSTRACT
OBJECTIVE: Since there are clinical and genetic differences between MS patients with intrathecal oligoclonal bands (OCB+) in the cerebrospinal fluid (CSF) compared with those without (OCB-), the aim was to find out if OCB- patients showed a different pattern of cytokine immune activation compared with OCB+ patients. METHODS: The study included 25 MS patients (10 OCB- and 15 OCB+) and 13 controls. A panel of cytokines was measured; IL-1ß, IL-6, IL-8/CXCL8, IL-10, TNF and GM-CSF in serum, CSF and in supernatants from polyclonally stimulated blood mononuclear cells, where also levels of IL-12p40, IL-13, IL-15, IL-17 and IFN-γ were measured. The concentrations of soluble (s) VCAM-1 and sCD14 were measured in serum and CSF. RESULTS: In general, there were no extensive differences in cytokine concentrations between the OCB- and OCB+ groups. CONCLUSION: OCB- MS patients do not seem to constitute a separate entity concerning inflammatory parameters measured as cytokine concentrations in CSF and blood.
Subject(s)
Cytokines/blood , Cytokines/cerebrospinal fluid , Inflammation Mediators/blood , Inflammation Mediators/cerebrospinal fluid , Multiple Sclerosis/immunology , Oligoclonal Bands/cerebrospinal fluid , Adult , Aged , Biomarkers/blood , Biomarkers/cerebrospinal fluid , Case-Control Studies , Cells, Cultured , Chi-Square Distribution , Female , Humans , Leukocytes, Mononuclear/immunology , Male , Middle Aged , Multiple Sclerosis/blood , Multiple Sclerosis/cerebrospinal fluid , Multiple Sclerosis/classification , Multiple Sclerosis/diagnosis , Regression Analysis , Sweden , Young AdultABSTRACT
We performed a candidate gene association study in 540 patients with primary Sjögren's Syndrome (SS) from Sweden (n=344) and Norway (n=196) and 532 controls (n=319 Swedish, n=213 Norwegian). A total of 1139 single-nucleotide polymorphisms (SNPs) in 84 genes were analyzed. In the meta-analysis of the Swedish and Norwegian cohorts, we found high signals for association between primary SS and SNPs in three gene loci, not previously associated with primary SS. These are the early B-cell factor 1 (EBF1) gene, P=9.9 × 10(-5), OR 1.68, the family with sequence similarity 167 member A-B-lymphoid tyrosine kinase (FAM167A-BLK) locus, P=4.7 × 10(-4), OR 1.37 and the tumor necrosis factor superfamily (TNFSF4=Ox40L) gene, P=7.4 × 10(-4), OR 1.34. We also confirmed the association between primary SS and the IRF5/TNPO3 locus and the STAT4 gene. We found no association between the SNPs in these five genes and the presence of anti-SSA/anti-SSB antibodies. EBF1, BLK and TNFSF4 are all involved in B-cell differentiation and activation, and we conclude that polymorphisms in several susceptibility genes in the immune system contribute to the pathogenesis of primary SS.
Subject(s)
OX40 Ligand/genetics , Protein-Tyrosine Kinases/genetics , Sjogren's Syndrome/genetics , Sjogren's Syndrome/immunology , Trans-Activators/genetics , B-Lymphocytes/immunology , Case-Control Studies , Cohort Studies , Female , Genetic Predisposition to Disease , Humans , Interferon Regulatory Factors/genetics , Interleukin-6/genetics , Lymphocyte Activation , Male , Middle Aged , Norway , Polymorphism, Single Nucleotide , STAT4 Transcription Factor/genetics , Sjogren's Syndrome/enzymology , SwedenABSTRACT
Studies on cytokine expression in blood cells are commonly performed on cryopreserved cells. Previous studies show that cryopreservation affects cytokine expression, but the findings are not consistent. This may be due to divergent effects of freezing on different cytokines, different stimuli, and different patient groups or to the use of different assays in the studies. This study was designed to investigate the effect of freezing on spontaneous, auto-antigen, allergen, and mitogen induced cytokine secretion from peripheral blood mononuclear cells from several groups of patients expressing different cytokine profiles; multiple sclerosis, atopic children, non-atopic children, and pregnant women. The expression of IFN-gamma, IL-4, IL-5, IL-9, IL-10, and IL-13 was analysed with ELISA, ELISPOT and/or real time RT-PCR. Our data provide evidence that the process of cryopreservation and thawing does affect the expression of cytokines, both at the protein and the mRNA level. Moreover, the effect varied among different cytokines, different stimuli, and different patient groups, which partly may be explained by differences in optimal freezing conditions for non-activated and activated cells. An increase of allergen and PHA stimulated IFN-gamma secretion in atopic children was found following cryopreservation, but no such increase in auto-antigen induced IFN-gamma was seen in MS-patients. The most consistent finding was that expression of IL-4 was generally decreased in spontaneous and auto-antigen/allergen induced expression in cryopreserved cells. In conclusion, this study points out the importance of investigation of the effects of freezing for each cytokine, stimuli and patient group before using frozen cells in studies of in vitro cytokine secretion.
Subject(s)
Blood Preservation/methods , Cryopreservation/methods , Cytokines/genetics , Cytokines/metabolism , Leukocytes, Mononuclear/immunology , Adult , Aged , Base Sequence , Child , DNA/genetics , Female , Gene Expression , Humans , Hypersensitivity, Immediate/blood , Hypersensitivity, Immediate/immunology , In Vitro Techniques , Interleukin-4/genetics , Interleukin-4/metabolism , Male , Middle Aged , Multiple Sclerosis/blood , Multiple Sclerosis/immunology , Polymerase Chain Reaction , Pregnancy , RNA, Messenger/genetics , RNA, Messenger/metabolism , Th1 Cells/immunology , Th2 Cells/immunologyABSTRACT
We used ELISPOT and cell ELISA to study secretion of IL-4, IFN-gamma, TGF-beta, IL-6, and TNF-alpha by circulating mononuclear cells during the course of Guillain-Barré syndrome (GBS). Compared to healthy controls, patients with GBS had higher numbers of TGF-beta-secreting cells and the number of individuals with myelin-peptide-induced IL-4 and TGF-beta secretion was higher in the GBS group. No significant differences were seen concerning the predominantly pro-inflammatory cytokines IFN-gamma, IL-6 or TNF-alpha. Our findings indicate a down-regulatory role for TGF-beta and IL-4 in GBS.
Subject(s)
Guillain-Barre Syndrome/immunology , Leukocytes, Mononuclear/metabolism , Transforming Growth Factor beta/biosynthesis , Adolescent , Adult , Aged , Female , Guillain-Barre Syndrome/blood , Humans , Interferon-gamma/analysis , Interleukin-4/analysis , Interleukin-4/biosynthesis , Interleukin-6/analysis , Lymphocyte Count , Male , Middle Aged , Tumor Necrosis Factor-alpha/analysisABSTRACT
The mechanisms behind the beneficial effects of interferon-beta1a (IFN-beta1a) and glatiramer acetate (GA) in the treatment of multiple sclerosis (MS) are still uncertain. Altered cytokine patterns have been suggested including inhibition of proinflammatory cytokines like interferon-gamma (IFN-gamma) and enhancement of anti-inflammatory cytokines such as interleukin-4 (IL-4). Twenty-nine patients with MS (10 untreated, nine treated with IFN-beta1a and 10 with GA) were investigated with elispot of peripheral blood mononuclear cells. Spontaneous and myelin induced (myelin basic protein (MBP), myelin oligodendrocyte glycoprotein (MOG)-14-39 and MOG 63-87) IFN-gamma, IL-4, IL-5 and IL-10 secretion was studied. We found a significant reduction of spontaneous IFN-gamma, IL-4 and IL-5, but no difference in IL-10 secreting cells in both groups of treated patients compared with the untreated patients. Myelin-specific responses showed a significant decrease of IFN-gamma and an increase of IL-5, but no change in IL-4 and IL-10 secreting cells in treated compared with untreated patients. Both treatment groups revealed similar cytokine secretion patterns except for a more pronounced decrease of both spontaneous and MOG 14-39 induced IL-4 secretion in the IFN-beta1a treated group. Thus, immunological effects of IFN-beta1a and GA were similar showing that disease promoting Th1 (IFN-gamma) cells were reduced while the potentially beneficial Th2 response (IL-4) was maintained.
Subject(s)
Adjuvants, Immunologic/therapeutic use , Cytokines/metabolism , Interferon-beta/therapeutic use , Multiple Sclerosis/drug therapy , Peptides/therapeutic use , Adult , Female , Glatiramer Acetate , Humans , Immunoassay , Interferon beta-1a , Interferon-gamma/metabolism , Interleukin-10/metabolism , Interleukin-4/metabolism , Interleukin-5/metabolism , Male , Middle Aged , Multiple Sclerosis/metabolism , Myelin Basic Protein/metabolism , Myelin Proteins , Myelin-Associated Glycoprotein/metabolism , Myelin-Oligodendrocyte GlycoproteinABSTRACT
Monoclonal expansion of B cells and plasma cells, producing antibodies against 'self' molecules, can be found not only in different autoimmune diseases, such as peripheral neuropathy (PN), but also in malignancies, such as Waldenström's macroglobulinaemia and B-type of chronic lymphocytic leukaemia (B-CLL), as well as in precancerous conditions including monoclonal gammopathy of undetermined significance (MGUS). About 50% of patients with PN-MGUS have serum antibodies against peripheral nerve myelin, but the specific role of these antibodies remains uncertain. The aims of the study were to establish, and characterize, myelin-specific B cell clones from peripheral blood of patients with PN-MGUS, by selection of cells bearing specific membrane Ig-receptors for myelin protein P0, using beads coated with P0. P0-coated magnetic beads were used for selection of cells, which subsequently were transformed by Epstein--Barr virus. The specificity of secreted antibodies was tested by ELISA. Two of the clones producing anti-P0 antibodies were selected and expanded. The magnetic selection procedure was repeated and new clones established. The cells were CD5+ positive, although the expression declined in vitro over time. The anti-P0 antibodies were of IgM-lambda type. The antibodies belonged to the VH3 gene family with presence of somatic mutations. The IgM reacted with P0 and myelin-associated glycoprotein (MAG), and showed no evidence for polyreactivity, in contrast to other IgM CD5+ clones included in the study as controls. The expanded clones expressed CD80 and HLA-DR, which is compatible with properties of antigen-presenting cells. The immunomagnetic selection technique was successfully used for isolation of antimyelin protein P0-specific clones. The cell lines may provide useful tools in studies of monoclonal gammopathies, leukaemia, and autoimmune diseases, including aspects of antigen-presentation by these cells followed by T cell activation.
Subject(s)
Antibodies, Monoclonal/biosynthesis , Autoantibodies/biosynthesis , Autoimmune Diseases of the Nervous System/immunology , B-Lymphocyte Subsets/immunology , Immunoglobulin M/biosynthesis , Myelin P0 Protein/immunology , Paraproteinemias/complications , Polyneuropathies/immunology , Aged , Antibodies, Monoclonal/immunology , Antibody Specificity , Antigen-Presenting Cells/immunology , Autoantibodies/immunology , Autoimmune Diseases of the Nervous System/etiology , B7-1 Antigen/analysis , Base Sequence , Cell Line, Transformed/immunology , Cells, Cultured/immunology , Clone Cells/immunology , Enzyme-Linked Immunosorbent Assay , Female , Gene Rearrangement, B-Lymphocyte, Heavy Chain , HLA-DR Antigens/analysis , Herpesvirus 4, Human/physiology , Humans , Immunoglobulin M/immunology , Immunoglobulin lambda-Chains/genetics , Immunomagnetic Separation , Immunophenotyping , Male , Middle Aged , Molecular Sequence Data , Paraproteinemias/immunology , Paraproteinemias/pathology , Polyneuropathies/etiology , Sequence Alignment , Sequence Homology, Nucleic AcidABSTRACT
Infections with the obligate intracellular bacterium Chlamydia trachomatis are characterized by avoidance of fusion between chlamydia-containing endosomes and lysosomes, bacterial persistence and development of post-infectious sequelae. In this report we show that C. trachomatis induces apoptosis in McCoy and HeLa cells. Apoptosis was monitored by three different techniques; enzyme-linked immunoassay (EIA) of fragmented nucleosomes, terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling (TUNEL) and flow cytometry of propidium iodide-stained cells. Apoptosis occurred in uninfected cells, was induced late in the chlamydial developmental cycle, beyond 24 h post-infection and was dependent on bacterial protein synthesis. Apoptosis was not significantly increased in infected, inclusion-containing cells. Treatment of cells with the antioxidants ascorbic acid (10 microM) and alpha-tocopherol (10 microM) reduced the degree of apoptosis. These results suggest that host cells infected with C. trachomatis generate proapoptotic stimuli that induce apoptosis in uninfected, neighbouring cells and that the redox state of the cell is a regulator in chlamydia-induced apoptosis.
Subject(s)
Apoptosis , Chlamydia trachomatis/pathogenicity , Reactive Oxygen Species , Animals , Antioxidants/pharmacology , Bacterial Proteins/biosynthesis , Cell Line , Humans , In Situ Nick-End Labeling , Mice , Oxidation-ReductionSubject(s)
Community Health Centers/statistics & numerical data , Emigration and Immigration , Health Services Needs and Demand , Translating , Adult , Aged , Clinical Competence , Community Health Centers/economics , Cultural Characteristics , Female , Health Services Needs and Demand/economics , Health Services Needs and Demand/statistics & numerical data , Humans , Male , Middle Aged , Referral and Consultation/economics , Referral and Consultation/statistics & numerical data , SwedenABSTRACT
A causal role of IL-4 (Th2) production for recovery in experimental allergic neuritis (EAN) was indicated by experiments where Th1-like autoreactive cell populations, taken from the induction phase of the disease, were deviated to extensive secretion of IL-4 in a selective fashion, by ex vivo stimulation with autoantigen in the presence of IL-4. The deviated cells were adoptively transferred to EAN rats at a time just prior to the onset of clinical signs. This treatment ameliorated EAN compared with sham treatment. This therapeutic approach, with generation of autoreactive IL-4-secreting cells ex vivo followed by subsequent adoptive transfer, may become a new selective treatment of organ-specific autoimmune diseases since, in contrast to previous attempts, it is done in a physiological and technically easy way.
Subject(s)
Adoptive Transfer/methods , Epitopes, T-Lymphocyte/immunology , Interleukin-4/biosynthesis , Myelin P2 Protein/immunology , Neuritis, Autoimmune, Experimental/immunology , Neuritis, Autoimmune, Experimental/prevention & control , Peptide Fragments/immunology , Th2 Cells/transplantation , Animals , Cytokines/metabolism , Epitopes, T-Lymphocyte/toxicity , Interleukin-4/genetics , Interleukin-4/pharmacology , Lymph Nodes/cytology , Lymph Nodes/immunology , Lymph Nodes/metabolism , Lymphocyte Transfusion , Male , Myelin P2 Protein/toxicity , Neuritis, Autoimmune, Experimental/metabolism , Peptide Fragments/toxicity , Rats , Rats, Inbred Lew , Recombinant Proteins/pharmacology , T-Lymphocyte Subsets/immunology , T-Lymphocyte Subsets/metabolism , T-Lymphocyte Subsets/transplantation , Th2 Cells/immunology , Th2 Cells/metabolismABSTRACT
PURPOSE: The purpose of this study was to investigate whether interleukin-1beta in synovial fluid or blood plasma is involved in the development of pain or hyperalgesia of the temporomandibular joint (TMJ), as well as reduced mandibular mobility and anterior open bite. PATIENTS AND METHODS: Twenty-nine patients with TMJ arthritis and seven healthy subjects were studied. VAS measurement of TMJ tenderness on palpation of the TMJ (TDP), TMJ pressure pain threshold and tolerance level (PPTL), mandibular mobility, pain during joint movements, and degree of anterior open bite (AOB) were assessed. IL-1beta levels were analyzed in TMJ synovial fluid (SF-IL-1beta) and blood samples and correlated with the preceding factors. RESULTS: SF-IL-1beta showed significant positive correlations with VAS measurement of pain, TDP, and AOB and a negative correlation with PPTL. CONCLUSIONS: This study indicates that IL-1beta in the synovial fluid is associated with pain and hyperalgesia in the TMJ region as well as an anterior open bite. Concerning the latter condition, IL-1beta seems to be a warning signal of tissue destruction.