Little is known about the immunology underlying variable treatment response in rheumatoid arthritis (RA). We performed large-scale transcriptome analyses of peripheral blood immune cell subsets to ...identify immune cells that predict treatment resistance.
We isolated 18 peripheral blood immune cell subsets of 55 patients with RA requiring addition of new treatment and 39 healthy controls, and performed RNA sequencing. Transcriptome changes in RA and treatment effects were systematically characterised. Association between immune cell gene modules and treatment resistance was evaluated. We validated predictive value of identified parameters for treatment resistance using quantitative PCR (qPCR) and mass cytometric analysis cohorts. We also characterised the identified population by synovial single cell RNA-sequencing analysis.
Immune cells of patients with RA were characterised by enhanced interferon and IL6-JAK-STAT3 signalling that demonstrate partial normalisation after treatment. A gene expression module of plasmacytoid dendritic cells (pDC) reflecting the expansion of dendritic cell precursors (pre-DC) exhibited strongest association with treatment resistance. Type I interferon signalling was negatively correlated to pre-DC gene expression. qPCR and mass cytometric analysis in independent cohorts validated that the pre-DC associated gene expression and the proportion of pre-DC were significantly higher before treatment in treatment-resistant patients. A cluster of synovial DCs showed both features of pre-DC and pro-inflammatory conventional DC2s.
An increase in pre-DC in peripheral blood predicted RA treatment resistance. Pre-DC could have pathophysiological relevance to RA treatment response.
To investigate metabolite alterations in the plasma of SLE patients to identify novel biomarkers and provide insight into SLE pathogenesis.
Patients with SLE (n = 41, discovery cohort and n = 37, ...replication cohort), healthy controls (n = 30 and n = 29) and patients with RA (n = 19, disease control) were recruited. Metabolic profiles of the plasma samples were analysed using liquid chromatography-time-of-flight mass spectrometry and capillary electrophoresis-time-of-flight mass spectrometry. Transcriptome data was analysed using RNA-sequencing for 18 immune cell subsets. The importance of histidine (His) in plasmablast differentiation was investigated by using mouse splenic B cells.
We demonstrate that a specific amino acid combination including His can effectively distinguish between SLE patients and healthy controls. Random forest and partial least squares-discriminant analysis identified His as an effective classifier for SLE patients. A decrease in His plasma levels correlated with damage accrual independent of prednisolone dosage and type I IFN signature. The oxidative phosphorylation signature in plasmablasts negatively correlated with His levels. We also showed that plasmablast differentiation induced by innate immune signals was dependent on His.
Plasma His levels are a potential biomarker for SLE patients and are associated with damage accrual. Our data suggest the importance of His as a pathogenic metabolite in SLE pathogenesis.
To describe the long-term clinical course of each manifestation of Behçet's disease (BD) and clarify factors involved in oral ulcer (OU) remission using clinical information of BD patients.
We ...retrospectively studied 155 BD patients visiting our hospital (1989-2020). We defined remission criteria for each manifestation and examined long-term clinical changes. Classification and regression trees and multivariable analyses were performed to investigate OU prognostic factors; hazard ratios were used to assign scores to prognostic factors deemed significant OU prognosis score (OuP score). Risk stratification was examined by dividing the OuP scores into four stages.
OUs appeared earliest, with the slowest decline in prevalence observed post-BD diagnosis. OU presence was the most common factor inhibiting complete remission. Young age at OU onset, never smoker, presence of genital ulcers, positive pathergy test, no usage of tumour necrosis factor inhibitors or of immunosuppressants, and long-term non-treatment or symptomatic treatment for OUs were poor OU prognostic factors. Based on multivariable analysis, the area under the curve of the OuP score to predict OU prognosis was 0.678.
Remission criteria for each symptom clarified that OU had the greatest impact on complete BD remission. Faster OU remission was associated with earlier OU therapeutic intervention other than symptomatic treatment.
Abstract
Background
Behçet’s syndrome (BS) is an immune-mediated disease characterized by recurrent oral ulcers, genital ulcers, uveitis, and skin symptoms. HLA-B51, as well as other genetic ...polymorphisms, has been reported to be associated with BS; however, the pathogenesis of BS and its relationship to genetic risk factors still remain unclear. To address these points, we performed immunophenotyping and transcriptome analysis of immune cells from BS patients and healthy donors.
Methods
ImmuNexUT is a comprehensive database consisting of RNA sequencing data and eQTL database of immune cell subsets from patients with immune-mediated diseases and healthy donors, and flow cytometry data and transcriptome data from 23 BS patients and 28 healthy donors from the ImmuNexUT study were utilized for this study. Differential gene expression analysis and weighted gene co-expression network analysis (WGCNA) were performed to identify genes associated with BS and clinical features of BS. eQTL database was used to assess the relationship between genetic risk factors of BS with those genes.
Results
The frequency of Th17 cells was increased in BS patients, and transcriptome analysis of Th17 cells suggested the activation of the NFκB pathway in Th17 cells of BS patients. Next, WGCNA was used to group genes into modules with similar expression patterns in each subset. Modules of antigen-presenting cells were associated with BS, and pathway analysis suggested the activation of antigen-presenting cells of BS patients. Further examination of genes in BS-associated modules indicated that the expression of
YBX3
, a member of a plasmacytoid dendritic cell (pDC) gene module associated with BS, is influenced by a BS risk polymorphism, rs2617170, in pDCs, suggesting that
YBX3
may be a key molecule connecting genetic risk factors of BS with disease pathogenesis. Furthermore, pathway analysis of modules associated with HLA-B51 indicated that the association of IL-17-associated pathways in memory CD8
+
T cells with HLA-B51; therefore, IL-17-producing CD8
+
T cells, Tc17 cells, may play a critical role in BS.
Conclusions
Various cells including CD4
+
T cells, CD8
+
T cells, and antigen-presenting cells are important in the pathogenesis of BS. Tc17 cells and
YBX3
may be potential therapeutic targets in BS.
The thermal environment map in street canyon is derived by using GIS building data and more detailed calculation, and its effectiveness is considered for implementing extreme high temperature ...measures. The influence of mean radiant temperature (MRT) is more dominant than the wind velocity on the distribution of standard new effective temperature (SET*) on the typical summer day in street canyon in the urban area of Kobe city, and the solar radiation shading is more effective in suppressing the rise of SET* in the daytime than improving the land coverage. The following strategy of extreme high temperature measures is derived by considering the thermal environment map in street canyon. Pedestrians may find the shaded places on the north-south road until 10:00 a.m. and after 3:00 p.m., due to the eastern building’s shade in the morning and the western building’s shade in the afternoon.
Previous gene expression analyses seeking genes specific to antineutrophil cytoplasmic antibody-associated vasculitis (AAV) have been limited due to crude cell separation and the use of microarrays. ...This study aims to identify AAV-specific gene expression profiles in a way that overcomes those limitations.
Blood samples were collected from 26 AAV patients and 28 healthy controls (HCs). Neutrophils were isolated by negative selection, whereas 19 subsets of peripheral blood mononuclear cells were sorted by fluorescence assisted cell sorting. RNA-sequencing was then conducted for each sample, and iterative weighted gene correlation network analysis (iterativeWGCNA) and random forest were consecutively applied to identify the most influential gene module in distinguishing AAV from HCs. Correlations of the identified module with clinical parameters were evaluated, and the biological role was assessed with hub gene identification and pathway analysis. Particularly, the module's association with neutrophil extracellular trap formation, NETosis, was analyzed. Finally, the module's overlap with GWAS-identified autoimmune disease genes (GADGs) was assessed for validation.
A neutrophil module (Neu_M20) was ranked top in the random forest analysis among 255 modules created by iterativeWGCNA. Neu_M20 correlated with disease activity and neutrophil counts but not with the presence of antineutrophil cytoplasmic antibody. The module comprised pro-inflammatory genes, including those related to NETosis, supported by experimental evidence. The genes in the module significantly overlapped GADGs.
We identified the distinct group of pro-inflammatory genes in neutrophils, which characterize AAV. Further investigations are warranted to confirm our findings as they could serve as novel therapeutic targets.
•Expression profile of a gene module in neutrophils characterizes AAV.•The module eigengene correlates with disease activity and neutrophil counts.•The module comprises pro-inflammatory genes, including those related to NETosis.•Genes in the module overlap autoimmune disease genes identified by GWAS.
The aim of this study was to investigate the effects of different practical training models on the comprehension and evaluation of practical training among dental students.
The study subjects were ...all sixth-year dental students at our institute, and the study took place over three consecutive years (n = 58, 63, and 65, respectively). In practical training, all students learned border molding, and practical models were modified each year from plaster models to silicone models and then to silicone models mounted in mannequins. Immediately after completing clinical training, all students were asked to complete questionnaires consisting of 21 items regarding their overall practical training and their clinical comprehension of border molding. All items were rated on a five-point Likert scale, and in order to reduce the large number of interrelated questions, exploratory factor analysis was carried out using maximum likelihood estimation with promax rotation (κ = 4) and Kaiser normalization. The number of factors was chosen using the Kaiser-Guttman rule, which states that the eigenvalue should be larger than 1, and the scree plot criteria. Items that scored less than 0.25 in communality and exhibited factor loading greater than 0.35 for more than one item were excluded. The defined factors were analyzed for the plaster models, the silicone models alone, and the silicone models with mannequins using the Kruskal-Wallis test and follow-up tests using Bonferroni-corrected Mann-Whitney U tests. The significance level was set at p < 0.05.
Exploratory factor analysis identified the following three factors: "knowledge of border molding"; "contents of practical training"; and "personal learning attitude". The students who used silicone models and mannequins gave significantly better evaluations on the "knowledge of border molding" (p < 0.001, both) and "contents of practical training" (p = 0.046, p < 0.001, respectively) subscales than those who used plaster models. No significant differences were observed between those who used silicone models and those who used mannequins. Moreover, no significant differences were found on the "personal learning attitude" subscale among students for any model.
The change in practical training models from plaster to silicone improved student evaluations of border molding training.
Objective
Idiopathic inflammatory myopathies (IIM) demonstrate characteristic clinical phenotypes depending on the myositis‐specific antibody (MSAs) present. We aimed to identify common or ...MSA‐specific immunological pathways in different immune cell types from peripheral blood by transcriptome analysis.
Methods
We recruited 33 patients with IIM who were separated into the following groups: 15 patients with active disease at onset and 18 with inactive disease under treatment. All patients were positive for MSAs: anti–melanoma differentiation‐associated gene 5 (MDA5) antibody (Ab) in 10 patients, anti‐Mi‐2 Ab in 7, and anti‐aminoacyl‐transfer RNA synthetase (ARS) Ab in 16. The patients were compared with 33 healthy controls. Twenty‐four immune cell types sorted from peripheral blood were analyzed by flow cytometry, RNA sequencing, and differentially expressed gene analysis combined with pathway analysis.
Results
The frequencies of memory B cell types were significantly decreased in active patients, and the frequency of plasmablasts was prominently increased in active patients with anti‐MDA5 Ab in comparison with healthy controls. The expression of type I interferon (IFN)‐stimulated genes of all immune cell types was increased in the active, but not inactive, patients. Endoplasmic reticulum stress‐related genes in all IIM memory B cells and oxidative phosphorylation‐related genes in inactive IIM double negative B cells were also increased, suggesting prominent B cell activation in IIM. Furthermore, active patients with anti‐MDA5 Ab, anti‐Mi‐2 Ab, or anti‐ARS Ab were distinguished by IFN‐stimulated and oxidative phosphorylation‐related gene expression in plasmablasts.
Conclusion
Unique gene expression patterns in patients with IIM with different disease activity levels and MSA types suggest different pathophysiologies. Especially, B cells may contribute to common and MSA‐specific immunological pathways in IIM.
To investigate the influence of wearing complete dentures on postural control in standing and walking.
Thirty-four edentulous patients participated in this study. All the subjects were wearing ...complete dentures, and the dentures were adjusted or replaced with new dentures when necessary. Measurements were performed under two conditions: wearing dentures and not wearing dentures. Standing stability was evaluated by the locus of center of mass, and gait stability was evaluated by the gait velocity, stride and gait cycle. In addition, gait stability was also evaluated by the maximum acceleration, maximum angle rate, lateral equilibrium, root mean square and harmonic ratio with a tri-axial accelerometer at a sampling rate of 66
Hz. Differences for the locus of center of mass, gait velocity, gait cycle and stride length were assessed with the paired
t test (
P
<
0.05). Other outcomes were compared with the Wilcoxon signed-rank test (
P
<
0.05).
With denture wear, the locus of center of mass was significantly shortened, and the gait velocity and harmonic ratio of the vertical angle rate were significantly increased; though other parameters showed no differences. Complete dentures produced an effect on the stability of edentulous patients under both static and dynamic conditions.
These results indicate that wearing complete dentures may be an effective aid to maintain and improve balance and control for elderly people.
Genetic studies have revealed many variant loci that are associated with immune-mediated diseases. To elucidate the disease pathogenesis, it is essential to understand the function of these variants, ...especially under disease-associated conditions. Here, we performed a large-scale immune cell gene-expression analysis, together with whole-genome sequence analysis. Our dataset consists of 28 distinct immune cell subsets from 337 patients diagnosed with 10 categories of immune-mediated diseases and 79 healthy volunteers. Our dataset captured distinctive gene-expression profiles across immune cell types and diseases. Expression quantitative trait loci (eQTL) analysis revealed dynamic variations of eQTL effects in the context of immunological conditions, as well as cell types. These cell-type-specific and context-dependent eQTLs showed significant enrichment in immune disease-associated genetic variants, and they implicated the disease-relevant cell types, genes, and environment. This atlas deepens our understanding of the immunogenetic functions of disease-associated variants under in vivo disease conditions.
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•Gene-regulation atlas of 28 immune cell types under immune-mediated diseases (IMDs)•Expression QTLs show immune cell-type and disease context specificity•Cellular pathways diversify eQTL effects under in vivo disease conditions•This atlas links IMD GWAS variants to susceptible genes, cell types, and environment
The gene-regulation atlas of 28 immune cell types was constructed with immune-mediated disease patient samples and shows the dynamics of gene regulation depending on cell types and immunological conditions.
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