Objective. To examine humoral and cellular immune responses induced by a live attenuated herpes zoster (HZ) vaccine in patients with rheumatoid arthritis (RA) compared with osteoarthritis (OA) patients. Methods. This was an observational study of a live attenuated HZ vaccine in 41 patients with RA receiving conventional disease-modifying antirheumatic drugs (cDMARD) and/or low-dose glucocorticoids (GC) and in 28 patients with OA. Blood samples were obtained before and at 12 weeks after HZ vaccination. Immunogenicity was assessed using varicella zoster virus (VZV)-specific interferon gamma ELISA and an in-house ELISA. Clinical outcomes, including adverse events, HZ occurrence, and RA flares, were analyzed. Results. No patients developed vaccination-induced HZ during the followup period (median = 1.6 yrs). The HZ vaccine induced a significant increase in the VZV-specific enzyme-linked immunospot spot-forming units and anti-VZV immunoglobulin G antibodies in patients with RA and OA. The number of spot-forming units was lower in patients with RA than in patients with OA both at baseline and at 12 weeks after vaccination. The disease activity index for patients with RA was similar at baseline and at 12 weeks after vaccination. However, 6 patients with RA (14.6%) experienced a flare during the 12 weeks. Overall, 17 (24.6%) participants reported a mild adverse event such as an injection site reaction (11.6%). Conclusion. The HZ vaccine induced VZV-specific cellular and humoral responses in patients with RA. Although patients with RA showed a weaker vaccine-induced VZV-specific cellular immune response than patients with OA, the vaccine may be considered in patients with RA receiving cDMARD and/or low dose GC.
Objective To examine whether a JAK inhibitor regulates functional responses of human salivary gland epithelial cells ( SGEC s) and disease parameters in an animal model of Sjögren's syndrome ( SS ). Methods Common differentially expressed genes ( DEG s) were analyzed among peripheral blood mononuclear cells from patients with primary SS and other data sets, using blood and SG tissue. Validation of expression in SG s was analyzed by focus score. Inhibition of messenger RNA expression of DEG s and BAFF by filgotinib was analyzed using reverse transcription–polymerase chain reaction in primary SGEC s. SG organoid cultures were used to determine the association between DEG s and BAFF via knockdown using small interfering RNA s or to determine regulation of BAFF by JAK inhibitor. Filgotinib (1.5 mg/kg) was intraperitoneally injected into 8‐week‐old NOD /ShiLtJ mice 3 times per week to analyze manifestations of disease. Finally, STAT signaling was assessed in human and mouse SGEC s. Results Expression of the DEG s IFNG and BAFF increased in SG s from patients with primary SS , as assessed by focus score. There was a significant correlation between IFIT 2 and BAFF expression. JAK inhibitor suppressed interferon ( IFN )–induced transcription of DEG s and BAFF in human primary SGEC s. Knockdown of DEG s or inhibition of JAK caused reduced secretion of BAFF in human SG organoid cultures. In addition, filgotinib‐treated mice exhibited increased salivary flow rates and marked reductions in lymphocytic infiltration of SG s. JAK inhibitor regulated IFN α‐ and IFN γ‐induced pSTAT ‐1 Y701 , pSTAT ‐3 Y705 , and protein inhibitor of activated STAT ‐3 ( PIAS ‐3) in human SGEC s as well as IFN γ‐induced pSTAT ‐1 Y701 , pSTAT ‐3 S727 , and PIAS ‐1 in mouse SGECs. Conclusion JAK inhibition controls aberrant activation of SGEC s and may be a novel therapeutic approach for primary SS .
Objective. Triggering receptor expressed on myeloid cells 1 (TREM-1), which amplifies the inflammation elicited by the Toll-like receptor pathway, was originally implicated in sepsis and bacterial infection. However, it has been suggested that TREM-1 may also play an important role in non-infectious inflammation. The present study was conducted to investigate whether TREM-1 is involved in human acute gouty inflammation. Methods. A total of 37 gout patients were recruited between March 2011 and January 2014 from Seoul St Mary's Hospital. The expression of TREM-1 on mononuclear cells was assessed using FACS analysis, immunostaining and real-time RT-PCR. To block the TREM-1 signal, soluble TREM-1 (sTREM-1) or the synthetic blocking peptide LP17 was used. The concentration of sTREM-1 was assessed by ELISA. Results. FACS analysis and real-time RT-PCR demonstrated that TREM-1 expression was higher in the SF mononuclear cells of acute gouty arthritis patients than in peripheral blood mononuclear cells (PBMCs). Immunohistochemical staining of tophi tissues revealed TREM-1 expression, with confocal microscopy demonstrating TREM-1 expression on tophi tissue macrophages. We also demonstrated that MSU treatment induced TREM-1 expression on the PBMCs of acute gout patients in vitro. Although blockade of TREM-1 did not directly suppress MSU-induced IL-1β production of PBMCs in vitro, the concentration of soluble TREM-1 was higher in the SF of gout vs OA patients and was positively correlated with serum CRP. Conclusion. TREM-1 is induced by MSU and is associated with the inflammation of human acute gouty arthritis.
Systemic lupus erythematosus (SLE) is an autoimmune disease in which the main contributors to organ damage are antibodies against autoantigens, such as double-stranded DNA (dsDNA). Calorie restriction and intermittent fasting (IF) have been shown to improve autoimmune disease symptoms in patients and animal models. Here, we tested the hypothesis that IF might improve symptoms in MRL/lpr mice, which spontaneously develop an SLE-like disease. Groups of mice were fed every other day (IF) or provided food ad libitum (controls), and various lupus-associated clinicopathological parameters were analyzed for up to 28 weeks. Contrary to expectations, anti-dsDNA antibody levels, immune complex deposition in the kidney, and glomerular injury were higher in the IF group than the control group, although there were no differences in spleen and lymph node weights between groups. Proteinuria was also worsened in the IF group. IF also increased the abundance of B cells, plasmablasts, and plasma cells and elevated autophagy in plasma cells in the spleen and lymph nodes. Secretion of anti-dsDNA antibody by splenocytes in vitro was reduced by chloroquine-induced inhibition of autophagy. These results suggest that IF exacerbates lupus nephritis in MRL/lpr mice by increasing autoantibody immune complex formation.
Abstract Sjögren’s syndrome (SS) is a chronic and systemic autoimmune disease characterized by lymphocytic infiltration in the exocrine glands. Although an association between inflammasome activation in peripheral blood cells and disease activity of SS has been reported, little is known about inflammasome activation in exocrine glands and its underlying mechanisms. We investigated the levels of AIM2, ASC, caspase-1, IL-1β, and IL-18 in salivary gland tissues and saliva from SS patients. We found that AIM2, ASC, Caspase-1, and IL-18 were increased in the saliva from primary SS patients compared with healthy and sicca controls. Numbers of AIM2-ASC speck also were elevated in the salivary gland epithelial cells of SS patients. The expression of caspase-1 correlated with type I interferon (IFN) signature gene expressions in the minor salivary glands (MSG) of SS patients, and was increased by type I IFN stimulation in salivary gland epithelial cells (SGECs). The AIM2 inflammasome was activated in SGECs after stimulation by the AIM2 ligand, poly(dA:dT). In addition, type I IFN accelerated AIM2 inflammasome activation as determined by the increased expression of caspase-1 in SGECs. In conclusion, the AIM2 inflammasome activation is increased in the SGECs of SS patients, and therefore should be considered an important pathogenic mediator of SS disease.
Background: Immune cells express the vitamin (vit) D receptor, and vit D is a potent immune-modulator.A negative correlation between serum vit D levels and rheumatoid arthritis (RA) disease activity has been reported.Therefore, we aimed to investigate if the sufficient serum vit D level is helpful to control disease activity in RA patients treated with interleukin (IL)-6 receptor antibody tocilizumab.Methods: RA patients taking tocilizumab were enrolled, and data were collected retrospectively.Disease activity scores (DAS) 28, serum vit D levels, modified Sharp scores of hand X-ray at the time of tocilizumab initiation, and follow-up data were analysed.Peripheral blood mononuclear cells were differentiated into T-helper (Th) 17 or osteoclasts in the presence of various concentrations of tocilizumab and/or 1,25(OH) 2 D. Th17 proportions were analysed by fluorescence-activated cell sorting.Supernatant cytokine levels were determined by enzyme-linked immunosorbent assay.Results: Among 98 RA patients taking tocilizumab, 34 (34.7%) had sufficient serum 25(OH) D levels (≥ 30 ng/mL) when tocilizumab was initiated.At 24 weeks, vit D sufficient patients had greater DAS28 reduction (64.6% ± 15.5% vs. 52.7%± 20.7%, P = 0.004), and lower disease activity (91.2% vs. 70.3%,P = 0.018) or remission (82.4% vs. 57.8%,P = 0.014).These differences in DAS28 reduction and the proportion of patients with remission persisted at 48 weeks.However, there was no significant difference in hand and wrist erosion progression.In vitro, tocilizumab and 1,25(OH) 2 D treatment synergistically suppressed IL-17 production and osteoclastogenesis.Conclusion: RA patients treated with IL-6 antibody show a better response when they have sufficient serum vit D. Tocilizumab and 1,25(OH) 2 D synergistically suppress IL-17 production and osteoclast differentiation in RA patients.
Metformin is originally introduced as a biguanide antidiabetic medication, has an anti-inflammatory effect via activating AMP-activated protein kinase (AMPK). Human adipose-derived stem cells (Ad-MSCs) are stromal cells derived from adipose tissue and known to have immunoregulatory activity. The study was undertaken to examine whether metformin-treated Ad-MSCs show more potent therapeutic effect in animal model of lupus and to clarify the underlying mechanism of potent immunoregulatory impact of metformin-treated Ad-MSCs.
Methods
To examine the effects of metformin, Ad-MSCs were incubated for 72 hour in the presence of metformin. Cellular phenotype of Ad-MSCs was analyzed by flow cytometry. Indoleamine 2,3-dioxygenase, IL-10 and TGF-1 expression was analyzed by real-time PCR and ELISA. AMPK-mTOR pathway was analyzed by Western blotting in Ad-MSCs with or without metformin. MRL/lpr mice weekly injected 1×10^6 metformin-treated Ad-MSCs in 0.1 ml PBS to lateral tail vein for 7 weeks. All mice were sacrificed at the age of 16 weeks. Urinary albumin-to-creatinine ratios were then calculated. The amount of anti-dsDNA IgG antibody in mice sera was measured by ELISA. PAS stained kidney sections were used for assessment of histology. Deposition of IgG and C3 was detected by confocal microscope. Population of cellular subset in spleen, kidney and blood was analyzed by Flow cytometry.
Results
In vitro, metformin-treated Ad-MSCs increased mRNA level of IDO, IL-10 and TGF-1 compared with untreated Ad-MSCs. Also, the concentrations of IDO, IL-10 and TGF- increased in culture supernatants. Metformin upregulated expression of p-AMPK and inhibited the expression of p-STAT3, p-mTOR, and p-Raptor in Ad-MSCs. Intravenously injected metformin-treated Ad-MSCs significantly reduced the splenomegaly and lymphadenopathy compared with untreated Ad-MSCs in MRL/lpr mice. In addition, Metformin-treated Ad-MSCs decreased anti-dsDNA antibodies in serum and proteinuria compared with untreated Ad-MSCs. Metformin-treated Ad-MSCs alleviated lupus nephritis, as judged by changes in the histopathology scores and immune complex deposition. Metformin-treated Ad-MSCs reduced CD90.2+T cells, CD90.2+CD4 CD8- (double negative) T cells, whereas CD4 +CD25+Foxp3+Treg cells were increased in splenocytes, kidney tissue and blood cells of MRL/lpr mice.
Conclusions
Metformin can optimize the immunomodulatory potential of Ad-MSCs, suggesting a promising strategy of MSC use in lupus treatment.
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