CD4(+) T cells of myasthenia gravis patients are characterized by ıncreased IL-21, IL-4, and IL-17A productions and higher presence of PD-1 and ICOS

Myasthenia gravis (MG) is an autoimmune disease mediated by autoantibodies predominantly against the acetylcholine receptor (AChR). Specific T cell subsets are required for long-term antibody responses, and cytokines secreted mainly from CD4(+) T cells regulate B cell antibody production. The aim of this study was to assess the differences in the cytokine expressions of CD4(+) T cells in MG patients with AChR antibodies (AChR-MG) and the effect of immunosuppressive (IS) therapy on cytokine activity and to test these findings also in MG patients without detectable antibodies (SN-MG). Clinically diagnosed AChR-MG and SN-MG patients were included. The AChR-MG patients were grouped as IS-positive and -negative and compared with age- and sex-matched healthy controls. Peripheral blood mononuclear cells were used for ex vivo intracellular cytokine production, and subsets of CD4(+) T cells and circulating follicular helper T (cTfh) cells were detected phenotypically by the expression of the chemokine and the costimulatory receptors. Thymocytes obtained from patients who had thymectomy were also analyzed. IL-21, IL-4, IL-10, and IL-17A productions in CD4(+) T cells were increased in AChR-MG compared to those in healthy controls. IS treatment enhanced IL-10 and reduced IFN-gamma production in AChR-MG patients compared to those in IS-negative patients. Increased IL-21 and IL-4 productions were also demonstrated in SN-MG patients. Among CD4(+) T cells, Th17 cells were increased in both disease subgroups. Treatment induced higher proportions of Th2 cells in AChR-MG patients. Both CXCR5(+) and CXCR5(-) CD4(+) T cells expressed higher programmed cell death protein 1 (PD-1) and inducible costimulatory (ICOS) in AChR-MG and SN-MG groups, mostly irrespective of the treatment. Based on chemokine receptors on CXCR5(+)PD-1(+) in CD4(+) T (cTfh) cells, in AChR-MG patients without treatment, the proportions of Tfh17 cells were higher than those in the treated group, whereas the Tfh1 cells were decreased compared with those in the controls. The relevance of CXCR5 and PD-1 in the pathogenesis of AChR-MG was also suggested by the increased presence of these molecules on mature CD4 single-positive thymocytes from the thymic samples. The study provides further evidence for the importance of IL-21, IL-17A, IL-4, and IL-10 in AChR-MG. Disease-related CD4(+)T cells are identified mainly as PD-1(+) or ICOS+ with or without CXCR5, resembling cTfh cells in the circulation or probably in the thymus. AChR-MG and SN-MG seem to have some similar characteristics. IS treatment has distinctive effects on cytokine expression.Istanbul Universit

IL-12 and IL-10 polymorphisms and their effects on cytokine production

Interleukin (IL)-12 is an inducer of differentiation of T helper (Th) cells towards Th1, whereas IL-10 is mainly an anti-inflammatory cytokine inhibiting Th1 functions. Single nucleotide polymorphisms (SNPs) in the regulatory sequences of genes are presumed to be associated with the differential production of cytokines. One IL12B 3′untranslated region (UTR) and five of IL10 gene promoter region SNPs were screened in 152 individuals by genotyping. IL-10 and IL-12 secretion of lipopolysaccharide (LPS), purified protein derivative (PPD) and Staphylococcus aureus Cowan strain I (SAC) stimulated peripheral blood mononuclear cells (PBMCs) was determined. The frequencies of the less frequent IL12B +16974 C, IL10 -2763 A -3575 A, -1082 G, -819 T and -592 A alleles were 27.4, 32.2, 25.9, 14.8, 9.3 and 8.6%, respectively. Individuals CC homozygous at IL12B 3′UTR had significantly higher IL-12 secretion levels from LPS and PPD stimulated PBMCs than AC heterozygotes (P = 0.03 and P = 0.02) or AA homozygotes (P = 0.02 and P = 0.05, respectively). IL10 -2763 and -3575 SNPs did not show any effect on in vitro secretion levels, whereas the association of proximal promoter -1082 SNP with IL-10 production is confirmed. IL10 proximal and distal promoter SNP distribution with estimated haplotype variations has implicated considerable similarities with the Caucasian populations in Turkey. © 2005 Elsevier Ltd. All rights reserved

Autoreactivity to myelin antigens related to HLA associations with multiple sclerosis

Multiple sclerosis (MS) is considered as an immune process influenced by genetic and environmental factors. HLA-DR2 and -DR4 have been documented to be associated with MS. The HLA-dependent differences of immune response to myelin and other antigens might point out some relevant mechanisms in MS development. The responses to myelin antigens and to PPD have been compared in 21 MS patients and 25 healthy controls (HCs) by primary proliferation and by short-term T-cell lines. There was a significantly higher response to MBP in DR2+ HCs compared to MS patients (SI: 5.9 versus 1.5, p=0.02). In short-term T-cell lines, we observed a higher response to PLP30-49 in both DR4+ HCs and MS patients. This response was significantly more frequent in DR4+ MS patients (34.6%) than both DR2+ MS patients (0%, p=0.0001) and DR4+ HCs (7.7%, p=0.001). The comparison between DR2+ and DR4+ MS patients has revealed that the response to MBP was also increased in DR4+ (p=0.02). Among DR4+ groups, an increased PPD response was detected in HCs compared to MS (65.2% versus 33.3%, p=0.01). These results may indicate that HLA-related differences to specific and recall antigens are detectable in MS and these differences may have implications in the disease pathogenesis

Serum cytokine profiles in Takayasu's arteritis: search for biomarkers

OBJECTIVES: Assessment of disease activity is one of the major difficulties in patients with Takayasu arteritis (TAK) during follow-up. To date, no biomarker is universally accepted to be a surrogate for active disease in TAK. In this study, we aimed to investigate levels of various pro-and anti-inflammatory molecules including serum granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin (IL)-6, IL-8, IL-10, IL-18 and IL-23 in patients with TAK.METHODS: The study included 51 patients (age: 40.6±12.2 years, F/M: 45/6) with TAK and 42 age- and sex-matched healthy controls (age: 38.1±7.4 years, F/M: 38/4). All patients fulfilled the criteria of the American College of Rheumatology (ACR). TAK patients were evaluated by physician's global assessment (PGA; active/inactive) and ITAS2010 (Indian Takayasu Arteritis Clinical Activity Score) in terms of clinical activity in baseline and follow-up visits. Commercial enzyme linked immuno-sorbent assay (ELISA) kits were used for measurements of serum cytokine levels.RESULTS: At baseline, 21 (41.2%) patients were active according to PGA and 8 (15.7%) according to ITAS2010. Serum IL-6, IL-8 and IL-18 levels were significantly higher in patients with TAK, whereas GM-CSF, IL-10, IL-23 levels were similar to healthy controls. IL-8 significantly decreased in the follow-up, associated with a decrease of clinical activity, whereas IL-23 level significantly increased. When assessed by ITAS2010 active patients had significantly higher IL-18 levels.CONCLUSIONS: We found significantly increased IL-6, IL-8 and IL-18 levels in patients with TAK compared to healthy controls. Only IL-18 level was significantly higher in active patients assessed by ITAS2010. IL-18 was also the only cytokine in our study that correlated with CRP. These findings suggest that cytokines associated with neutrophilic, pro-inflammatory responses such as IL-6, IL-8 and IL-18 can be potential biomarkers for the assessment of disease activity in TAK and warrant further studies in larger series

Cytokines and chemokines in neuro-Behçet's disease compared to multiple sclerosis and other neurological diseases.

Cytokines and chemokines in cerebrospinal fluid (CSF) can have implications on the pathogenesis of neuro-Behcet's disease (NB). CSF and serum samples from 33 patients with NB, 25 with multiple sclerosis (MS), 20 patients with infectious and/or inflammatory neurological diseases (IN) and 14 with other noninflammatory neurological diseases (NIN) were investigated by ELISA. In the CSF, CXCL10 levels were significantly higher in NB and IN than NIN and MS, whereas CXCL8 was increased in NB compared to NIN. CCL2 levels in MS CSF and sera were lower, whereas CXCL8 in MS sera was higher than the other groups. IL-12 was elevated in CSF of IN compared to NB and NIN and also in the CSF of MS compared to NIN. No difference was detected for IL-10 and IL-17. These results reflect that NB has a mediator pattern in resemblance with non-specific inflammations such as neuro-infections compared to autoimmune disorders such as multiple sclerosis, suggesting that a currently unknown infection might be the trigger of a vasculitic process in the central nervous system (CNS). (C) 2003 Elsevier B.V All rights reserved

Intrathecal oligoclonal IgG bands are infrequently found in neuro-behçet's disease

Objectives. Oligoclonal bands (OCB) of immunoglobulins (IgG) in the cerebrospinal fluid (CSF) provides an evidence for the humoral response and have been screened in the CSF and serum of patients revealing 5 different patterns. In this study, patients with Behçet's disease (BD) are screened in a larger sample to potentially provide information about the possible role of CSF oligoclonal immunoglobulins in the diagnosis of this disease. Methods. Paired CSF and serum samples from 121 consecutive BD patients with neurological complaints (43 women and 78 men) were included in this study. Parenchymal NBD was diagnosed in 74 patients, and 22 patients had cerebral venous sinus thrombosis (CVST); of the remaining patients, 18 had primary headache disorders not directly associated with BD, and 7 had a cerebrovascular event. OCB of IgG were detected by isoelectric focusing on agarose and immunoblotting of matched serum and CSF sample pairs. Intrathecal production of IgG only is considered positive (Pattern 2 or 3). Results. In the whole group, only 8 patients had OCB in the CSF showing pattern 2. All these positive cases had parenchymal neuro-BD (10.8% positive and 78.4% negative in parenchymal neuro-BD group). All other groups were negative. Conclusion. The rare presence of oligoclonal IgG bands in CSF can be utilised as another laboratory finding in the diagnosis of NBD. © Clinical and Experimental Rheumatology 2013

Plasma pentraxin-3 levels in patients with Takayasu's arteritis during routine follow-up

© Clinical and Experimental Rheumatology 2016.Objective. To date, no biomarker is universally accepted to be a surrogate for active disease being one of major difficulties in follow-up of Takayasu's arteritis (TAK). In this study, we aimed to investigate plasma pentraxin-3 (PTX-3) levels and its correlation with activity in patients with TAK. Methods. This cross-sectional study included 94 patients (age: 43.3±13.6 years, F/M: 80/14) with TAK, 40 age-sex matched control donors (age: 41.5±9.3 years, F/M: 28/12). TAK patients were evaluated by physician's global assessment (PGA; active/inactive), as well as with the activity definition by Kerr et al. and with a new composite index of ITAS2010 (Indian Takayasu Clinical Activity Score). Plasma PTX-3 levels are measured with an enzyme linked immunosorbent assay kit. Results. Thirty-three (35.5%) patients were clinically active with PGA, while 25 (31.6%) patients and 28 (31.8%) patients were accepted to have active disease according to Kerr activity criteria and ITAS2010, respectively. Plasma PTX-3 levels were significantly higher in TAK patients compared to healthy controls (3.5±2.5 ng/ml vs. 2.5±1.6 ng/ ml, p=0.029). However, PTX-3 levels were similar among active and inactive patients according to all three assessment tools. PTX-3 levels significantly correlated only with serum CRP levels. Conclusion. Although plasma PTX-3 levels were higher in patients with TAK compared to healthy controls, we observed no association with disease activity, limiting the role of PTX-3 level as a biomarker for active disease in TAK

Effect of mild heat stress on heat shock protein 70 in a balneotherapy model

Introduction: Fever-induced mild heat stress plays an important role in cellular responses. The best-known heat shock response is heat shock protein (HSP) secretion. We investigated the effect of mild heat stress on serum HSP70 levels and a possible association between the immune system and these molecules by applying a mild heat stress model using a thermal water bath

Increased costimulatory molecule expression of thymic and peripheral B cells and a sensitivity to IL-21 in myasthenia gravis.

B cells may contribute to the pathogenesis of myasthenia gravis with anti-acetylcholine antibodies (AChR + MG) by co-stimulation or selection of T cells

Anti-a alpha B-crystallin immunoreactivity in inflammatory nervous system diseases

alphaB-Crystallin, a small heat shock protein, is an immunodominant antigen with increased tissue expression in demyelination. To investigate the humoral response against alphaB-crystallin, the sera and CSF samples of patients with multiple sclerosis (MS), Guillain-Barre syndrome (GBS), neuro-Behcet's disease (NBD) and other non-inflammatory neurological diseases (NIND) were screened by enzyme-linked immunosorbent assay for anti-alphaB-crystallin IgG and IgM antibodies. Serum and CSF IgG antibody responses to alphaB-crystallin were significantly elevated only in NBD patients (serum IgG, NBD 1.29+/-0.49 vs. NIND 0.95+/-0.39, P=0.01; CSF IgG, NBD 1.22+/-0.64 vs. NIND 0.81+/-0.35, P=0.01). Similarly, high serum IgM antibody titres were also detected in NBD (1.83+/-0.72 vs. 1.16+/-0.49, P=0.0005) and in MS (1.57+/-1.07, P=0.046), whereas elevated CSF IgM responses were observed only in GBS (2.09+/-1.09 vs. 1.41+/-0.7, P=0.007). Humoral responses against alphaB-crystallin are increased in NBD and GBS, which may implicate this central nervous system antigen in the causation and pathogenesis of these inflammatory nervous system disorders