This study investigated the association between intestinal microbiota abundance and diversity and cluster of differentiation (CD)4+ T cell subpopulations, cytokine levels, and disease activity in rheumatoid arthritis RA. A total of 108 rheumatoid arthritis (RA) patients and 99 healthy control (HC) subjects were recruited. PICRUSt2 was used for functional metagenomic predictions. Absolute counts of peripheral CD4+ T cell subpopulations and cytokine levels were detected by flow cytometry and with a cytokine bead array, respectively. Correlations were analyzed with the Spearman rank correlation test. The results showed that the diversity of intestinal microbiota was decreased in RA patients compared to HCs. At the phylum level, the abundance of Firmicutes, Fusobacteriota, and Bacteroidota was decreased while that of Actinobacteria and Proteobacteria was increased and at the genus level, the abundance of Faecalibacterium, Blautia, and Escherichia-Shigella was increased while that of Bacteroides and Coprococcus was decreased in RA patients compared to HC subjects. The linear discriminant analysis effect size indicated that Bifidobacterium was the most significant genus in RA. The most highly enriched Kyoto Encyclopedia of Genes and Genomes pathway in RA patients was amino acid metabolism. The relative abundance of Megamonas, Monoglobus, and Prevotella was positively correlated with CD4+ T cell counts and cytokine levels; and the relative numbers of regulatory T cells (Tregs) and T helper (Th17)/Treg ratio were negatively correlated with disease activity in RA. These results suggest that dysbiosis of certain bacterial lineages and alterations in gut microbiota metabolism lead to changes in the host immune profile that contribute to RA pathogenesis.

Growing experimental and clinical evidence suggests that a chronic inflammatory response induced by gut microbiome critically contribute to the development of rheumatoid arthritis (RA). Previous studies demonstrated the disturbance of lymphocyte subpopulations in RA patients. The purpose of this study was to explore the characteristics of gut microbiome and the associations between bacterium and lymphocyte subpopulations as well as cytokines in patients with RA. Fecal samples from 205 RA patients and 199 healthy controls (HCs) were collected for bacterial DNA extraction and 16S ribosomal RNA (rRNA) gene sequencing. The levels of peripheral lymphocyte subpopulation such as T, B, CD4+T, CD8+T, NK, T helper 1 (Th1), Th2, Th17, and regulatory T cells (Tregs) of these subjects were detected by flow cytometry combined with standard absolute counting beads. The serum levels of cytokines interleukin-2 (IL-2), IL-4, IL-6, IL-10, IL-17, tumour necrosis factor-α (TNF-α), and interferon-γ (INF-γ) were tested by flow cytometric bead array (CBA). Alpha and beta diversity of gut microbiome were explored by bioinformatics analysis. Spearman rank correlation test was used to explore the relationships between gut microbiome and lymphocyte subsets as well as serum cytokines. The diversity and relative abundance of intestinal microbiota in patients with RA were significantly different from those in HCs. Detailly, the abundant of phylum Proteobacteria in RA patients was more than that in HCs, while Firmicutes was less than in HCs. There was increased relative abundance of genus Clostridium_XlVa as well as genus Blautia, more abundance of Ruminococcus2 in patients with lower levels of T, B, CD4+T, and Tregs. In addition, the relative abundances of Pelagibacterium, Oxalobacter, ClostridiumXlVb, and ClostridiumXVIII were correlated with cytokines. Gut microbiome of RA patients was clearly different from that of HCs. Abnormal bacteria communities are associated with the altered levels of lymphocyte subpopulation and cytokines, which might be one of the pathogenesis of RA.

Mo.), was assessed for the rapid detection of group A beta-hemolytic streptococcus directly from a throat swab. Four hundred and thirty-five throat swab specimens from children with suspected group A streptococcal infection were tested by the Ten-Minute Group A Strep ID and by concurrent conventional culture. The strep test was effective in detecting group A streptococcal infection; 90% (63/70) of culture-positive specimens gave positive latex tests. The specificity of the test was 99.2% (362/365). The predictive values were 95.5% for positives and 98.1% for negatives. Overall agreement with culture was 98 % (425/435). This test offers a sensitive and specific method for the early detection of group A beta-hemolytic streptococci in throat swabs of children and would be most useful in a hospital laboratory or pediatrician's office.

Systemic lupus erythematosus (SLE) characterized by immune dysfunction is possibly more vulnerable to herpes simplex virus (HSV) infection. The infection has been intensively considered a common onset and exacerbation of SLE. This study is aimed at elucidating the causal association between SLE and HSV. A bidirectional two‐sample Mendelian Randomization (TSMR) analysis was systematically conducted to explore the causal effect of SLE and HSV on each other. The causality was estimated by inverse variance weighted (IVW), MR‐Egger and weighted median methods based on the summary‐level genome‐wide association studies (GWAS) data from a publicly available database. Genetically proxied HSV infection exhibited no causal association with SLE in the forward MR analysis using IVW method (odds ratio [OR] = 0.987; 95% confidence interval [CI]: 0.891–1.093; p = 0.798), nor did HSV‐1 IgG (OR = 1.241; 95% CI: 0.874–1.762; p = 0.227) and HSV‐2 IgG (OR = 0.934; 95% CI: 0.821–1.062; p = 0.297). Similar null results with HSV infection (OR = 1.021; 95% CI: 0.986–1.057; p = 0.245), HSV‐1 IgG (OR = 1.003; 95% CI: 0.982–1.024; p = 0.788) and HSV‐2 IgG (OR = 1.034; 95% CI: 0.991–1.080; p = 0.121) were observed in the reverse MR where SLE served as the exposure. Our study demonstrated no causal association between the genetically predicted HSV and SLE.

Objectives To leverage the high clinical heterogeneity of systemic lupus erythematosus (SLE), we developed and validated a new stratification scheme by integrating genome-scale transcriptomic profiles to identify patient subtypes sharing similar transcriptomic markers and drug targets. Methods A normalized compendium of transcription profiles was created from peripheral blood mononuclear cells (PBMCs) of 1046 SLE patients and 86 healthy controls (HCs), covering an intersection of 13 689 genes from six microarray datasets. Upregulated differentially expressed genes were subjected to functional and network analysis in which samples were grouped using unsupervised clustering to identify patient subtypes. Then, clustering stability was evaluated by the stratification of six integrated RNA-sequencing datasets using the same method. Finally, the Xgboost classifier was applied to the independent datasets to identify factors associated with treatment outcomes. Results Based on 278 upregulated DEGs of the transcript profiles, SLE patients were classified into three subtypes (subtype A-C) each with distinct molecular and cellular signatures. Neutrophil activation-related pathways were markedly activated in subtype A (named NE-driving), whereas lymphocyte and IFN-related pathways were more enriched in subtype B (IFN-driving). As the most severe subtype, subtype C [NE-IFN-dual-driving (Dual-driving)] shared functional mechanisms with both NE-driving and IFN-driving, which was closely associated with clinical features and could be used to predict the responses of treatment. Conclusion We developed the largest cohesive SLE transcriptomic compendium for deep stratification using the most comprehensive microarray and RNA sequencing datasets to date. This result could guide future design of molecular diagnosis and the development of stratified therapy for SLE patients.

Introduction Osteoporosis (OP) is one of the major comorbidities of rheumatoid arthritis (RA). Recent studies have shown that immune cells modulate bone health and regulate bone remodeling. However, the alterations of lymphocyte subsets in RA patients with OP are unclear. Here, we assessed the absolute numbers and proportions of the subsets in RA sufferers with OP and investigated the clinical significance. Methods A total of 777 RA patients and 117 gender- and age-matched healthy controls (HCs) were enrolled in this study. Patients were divided into RA-non-OP and RA-OP group according to their bone mineral density (BMD) and the history of fragility fracture. Peripheral lymphocyte subsets of participants were assessed by flow cytometry. Results Among 220 (28.31%) RA-OP patients, there were higher levels of erythrocyte sedimentation rate (ESR) ( P = 0.011), C-reactive protein (CRP) ( P = 0.028), rheumatoid factor (RF) ( P = 0.013) and anti-cyclic citrullinated peptide antibody (ACPA) ( P = 0.010), while red blood cells (RBC) ( P = 0.039) were lower than those in RA-non-OP group. Compared with those of HCs and RA-non-OP group, the level of circulating Th17 cells in RA-OP patients was significantly increased ( P < 0.05), while those of Tregs decreased ( P < 0.01), leading to a higher ratio of Th17/Treg ( P < 0.01). Notably, the level of B cells in both RA-non-OP and RA-OP group was reduced, this alteration was more obvious in patients with OP ( P < 0.05). Conclusions Immune disorders characterized by peripheral Th17/Treg imbalance and reduced B cells may contribute directly or indirectly to OP in RA, and this deserves more clinical attention. Supplementary Information The online version contains supplementary material available at 10.1007/s40744-022-00452-x.

Objective The most used drug in rheumatoid arthritis (RA) remains methotrexate (MTX). Unfortunately, up to 50% of patients do not achieve a clinically adequate outcome. Here we study whether the gut microbiota patterns can aid in the prediction of MTX efficacy in RA. Method To dissect gut microbiome profiles of RA patients (n = 145), 16S rRNA gene sequencing was performed. Dirichlet multinomial mixture (DMM) clustering was used to identify enterotypes at genus level. The relationships between enterotypes and clinical measures (such as lymphocyte subsets and cytokines detected by flow cytometry) were explored. Then, enterotype stability was evaluated by the stratification of the RA patients cohort in Shanghai, China (n = 66) using the same method. Finally, the enterotype-based gut microbial human index (EGMI) classifier was applied to another independent RA patients cohort (n = 27) to identify the factors associated with MTX clinical response. Results Our analysis revealed that the RA patients always displayed two different dysbiotic microbiota patterns: RA E1 comprised predominantly Prevotella and RA E2 comprised predominantly Bacteroides. Among all of the lymphocyte subsets and cytokines, only the number of CD8+ T cells showed a significant difference between RA E1 and RA E2. These results were validated in the RA patients cohort in Shanghai, China. Significant associations of RA E1 with clinical response to subsequent MTX treatment were confirmed by another independent RA patients cohort. Conclusion Together, EGMI classifier was useful to identify precisely and effectively enterotypes of individual RA patients, which could effectively evaluate MTX clinical responses.

Background:Systemic lupus erythematosus (SLE) is a multisystem autoimmune disease characterized by abnormal activation of circulating lymphocytes and overproduction of autoantibodies1. Breakdown of self-tolerance is considered as a critical cause in the development of SLE2. However, the quantitative changes of lymphocyte subsets in SLE are unclear. Since low-dose IL-2 and several drugs have been used to promote the proliferation of regulatory T cells (Tregs)3, we developed immunoregulatory therapies using these drugs to rebalance effector T cells with Tregs and test whether they are benefit to remission disease activity of SLE.Objectives:To observe the different levels of peripheral lymphocyte subsets at the first laboratory examination of SLE patients with those of healthy controls (HCs) and to evaluate the effect of immunoregulatory combination therapies on levels of lymphocyte subsets in patients with SLE.Methods:From September 2014 to December 2019, a total of 985 diagnosed patients with SLE (878 females, 107 males, mean age 42.99±13.37 years) and 206 healthy adults were enrolled in this retrospective cross-sectional study. And 795 patients with SLE (711 females and 84 males, mean age 38.26±15.242 years) were received the immunomodulatory drugs (IMiDs) such as low-dose interleukin-2, rapamycin, metformin, retinoic acid, coenzymes Q10 or other immunomodulatory treatments. The absolute numbers of T, B, NK, CD4+T, CD8+T, Th1, Th2, Th17 and CD4+CD25+Foxp3+T regulatory cells (Tregs) in peripheral blood (PB) of these individuals were measured by Flow Cytometer (FCM) combined with standard absolute counting beads.Results:As compared with those of HCs, patients with SLE had lower absolute numbers of total T, NK, and CD4+T but higher proportions of all lymphocyte subpopulations except NK, CD4+T cells(P< 0.001) (Figure 1 A, C). Notably, the absolute numbers and proportions of Tregs as well as Th1 in CD4+T subsets were decreased (P<0.05) (Figure 1 B, D). Further, there was a significant increase in the ratio of Teffs/Tregs such as Th1/Tregs, Th2/Tregs and Th17/Tregs (P<0.05) (Figure 1 E). After receiving immunoregulatory combination therapies, the absolute numbers and proportions of T, NK, CD4+T, and CD8+T were increased, while the proportion of B cells was decreased (Figure 2 A, C); the absolute numbers of most CD4+T subsets as well as the proportions of only Th1 and Tregs were significantly increased (P< 0.001) (Figure 2 B, D). The ratios of Th1/Th2 and Th1/Tregs increased while that of Th17/Tregs and Th2/Tregs decreased (P<0.01) (Figure 2 E).Conclusion:Quantitative and functional alterations of peripheral lymphocyte subsets, especially reduced Tregs, play crucial roles in the pathogenesis of the patients. Immunoregulatory combination therapies mainly promote the proliferation and functional recovery of Tregs to rebalance pro- and anti-inflammatory T cells in patients with SLE for patients’ symptoms remission.References:[1]Sharabi A, Tsokos GC. T cell metabolism: new insights in systemic lupus erythematosus pathogenesis and therapy. Nat Rev Rheumatol 2020 doi: 10.1038/s41584-019-0356-x [published Online First: 2020/01/18][2]Durcan L, O’Dwyer T, Petri M. Management strategies and future directions for systemic lupus erythematosus in adults. Lancet 2019;393(10188):2332-43. doi: 10.1016/S0140-6736(19)30237-5 [published Online First: 2019/06/11][3]Spolski R, Li P, Leonard WJ. Biology and regulation of IL-2: from molecular mechanisms to human therapy. Nat Rev Immunol 2018;18(10):648-59. doi: 10.1038/s41577-018-0046-y [published Online First: 2018/08/10]Acknowledgments :None.Disclosure of Interests:None declared