Abstract Rationale People with Cystic Fibrosis (CF) often experience acute clinical deteriorations known as pulmonary exacerbations (PEx). PEx can be associated with increased airway inflammation, leading to lung function decline and mortality. Untreated females with CF experience more frequent and severe PEx than their male counterparts, suggesting sex-specific pathogenesis. Using single-cell RNA sequencing (scRNAseq), we previously identified CF-specific impairments in host defense and proinflammatory gene expression in immature airway monocytes. Prior analysis demonstrated sex-specific pulmonary neutrophil transcripts in people with CF who had frequent vs. infrequent PExs. Here, we sought to determine if there are sex-specific transcripts in monocytes of patients with CF who experience frequent vs. infrequent exacerbations. Methods We conducted a cross-sectional study of 31 sputum samples collected from untreated adults with CF at the Yale Adult CF Program (13 male [M], 18 female [F]) over the course of 2 years. Following sputum induction, we generated cDNA libraries from single cells following our previously established scRNAseq protocols. Sequencing was completed at the Yale Genomics and Bioinformatics Core. We defined four analysis groups based on sex (M, F) and the frequency of patient exacerbations per year, characterized as two or more per year (high exacerbators, HE) or less than two per year (low exacerbators, LE). We compared differentially expressed genes (DEG) by group while adjusting for a false discovery rate (FDR) of 0.05 using a standardized pipeline with the R package Seurat 5.1.0. Results Volcano plots were used to visualize gene expression differences in monocytes between HE and LE groups for both males and females. HE females displayed increased expression of the proinflammatory genes CCL20, HIF1A, and RHOA (FC >3.2, p < 10-16), however multiple proinflammatory genes were downregulated (CXCL9, ALCAM, CCL3, FC <-0.54, p < 10-5), suggesting a dysfunctional inflammatory transcriptional program. HE males demonstrated increased expression of several proinflammatory genes (e.g., CXCL10, CCL2, CCL4L2, FC>1.6, p < 0.001), which are distinct from the upregulated proinflammatory genes in HE females (Figure 1). Conclusion These findings suggest that sex- and cell-specific transcriptional abnormalities are associated with PEx frequency in patients with CF and contribute to sexually dimorphic clinical presentations. These subgroup-specific DEGs show promise for the development of personalized therapeutic approaches to reduce PEx frequency and narrow the sex gap in sexually dimorphic features of CF. This abstract is funded by: CFF, NIH

Common and rare genetic variants in leucine-rich repeat kinase 2 (LRRK2) have been linked with sporadic and familial Parkinson's disease (PD). Recently, we discovered that common genetic variation near the LRRK2 locus determined survival in progressive supranuclear palsy (PSP). Our study aimed to explore biomarkers of LRRK2 and lysosomal dysfunction in PSP. Immunoblotting was used to measure total LRRK2 and LRRK2-dependent Rab10 phosphorylation at the threonine 73 residue (pRab10Thr73) in neutrophil and monocyte samples from PSP and control participants. Urine samples were applied to a multiplexed assay to quantitate bis(monoacylglycerol)phosphate (BMP) species as markers of lysosomal dysfunction. Cerebrospinal fluid (CSF) samples from a wider cohort of PSP and control participants were applied to a stable isotope standards and capture by anti-peptide antibodies assay to measure total LRRK2 and pRab10Thr73 levels. LRRK2 genotypes (rs76904798 and rs2242367) and 1-year change in Progressive Supranuclear Palsy Rating Scale (PSPRS) scores were obtained. A total of 61 PSP and 34 control participants were included. Total urine 22:6-BMP levels were higher in PSP versus control samples (P = 0.04) and correlated with CSF total LRRK2 levels (r = 0.49, P = 0.04). There were no group-level differences in monocyte and CSF levels of total LRRK2 and pRab10Thr73. In PSP, carriers of the alternate allele (CT and TT genotypes) at the LRRK2 PD risk variant, rs76904798, had higher levels of CSF total LRRK2 versus CC genotype (P = 0.02). Baseline monocyte total LRRK2 levels predicted 1-year change in the PSPRS score (P = 0.008). Biochemically defined lysosomal dysfunction is evident in PSP. Genetic and biochemical stratification may identify PSP patients that would benefit from LRRK2-targeting therapies. © 2026 The Author(s). Movement Disorders published by Wiley Periodicals LLC on behalf of International Parkinson and Movement Disorder Society.

Abstract Description E-cigarette (e-cig) use has increased substantially but the health effects are poorly understood. E-cig vapor contains nicotine like tobacco cigarette smoke, which is known to increase the risk of cardiovascular disease (CVD), in part via monocyte activation. Functional differences in monocytes from males and females with CVD have been reported. Thus, we hypothesized that e-cigarette exposure activates monocytes, with a greater effect in females. We recruited 10 e-cig users (EC; 5F/5M) and 12 healthy controls (HC; 7F/5M) with no history of e-cig use, quantified circulating monocyte subsets, and performed bulk ATACseq and RNAseq on purified CD14+CD16- classical monocytes (CM). Compared to HC, EC had lower frequencies of CM (p < 0.05) and higher frequencies of CD14-CD16+ nonclassical monocytes (NCM p < 0.05), but no sex differences were observed. ATACseq identified 16,508 differentially accessible chromatin regions (DAR) in female EC vs HC and 8,744 DAR in female vs male EC. Of these, over 60% of these are located near the promoter region. Additionally, GeneOntology analysis revealed enriched pathways in female HC associated with myeloid cell differentiation and antigen processing. RNAseq analysis identified altered gene expression of functional monocyte genes. Our results indicate NCM are increased in female EC, which has been associated with CVD. Further, functional differences in monocytes from female EC may become apparent with maturation or interactions with the endothelium. Funding Sources Supported by NIH T32AI007485: NIH ES005605-32; University of Iowa Lulu Merle Fellowship, University of Iowa Inflammation Program Topic Categories Innate Immune Responses and Host Defense: Cellular Mechanisms (INC)

Abstract Description Chronic myelomonocytic leukemia (CMML) is considered to be a chronic myelodysplastic and myeloproliferative neoplasm characterized by a persistent increase in abnormal monocytes. According to the 2022 WHO classification,CMML is defined when 94% and more CD14+CD16- cells are detected in monocytes by flow cytometry. However more than 90% of those cells are also detected in patients with reactive monocytosis (RM), this makes the differentiation of CMML from RM difficult. We investigated the differences in cell morphology and compared the values ??of monocyte parameters side fluorescence (SFL) obtained by flow cytometry between both diseases. In this study, five patients with CMML and 44 patients with RM were enrolled. In morphological comparison of monocytes, the size of monocytes from patients with RM (20.39 ± 2.24 mm) was significantly bigger than that from CMML (17.25 ± 1.02 mm). When comparing the number of vacuoles over 1.0 mm, 3.3 ± 1.0 vacuoles were detected in monocytes from patients with RM and 1.0 ± 0.3 vacuoles were detected in monocytes from CMML. When the mean gray values of monocytes were examined, values of monocytes from RM (156.1 ± 3.7) were significantly higher than those from CMML (145.9 ± 3.7). The median values of SFL in monocytes by flow cytometry were 103.4 ± 5.8 (RM) and 114.1 ± 4.6 (CMML), SFL in monocytes from CMML was significantly higher than that from RM. Morphological characteristics and SFL may be useful in the differentiation of CMML from RM. Funding Sources Supported by JSPS KAKANHI, JP 21K15645 Topic Categories Lymphocyte Differentiation and Peripheral Maintenance (LYM)

Recent evidence increasingly supports a potential role of Perivascular Macrophages (PVMs), a unique subpopulation of brain immune cells, in the pathogenesis of Alzheimer's disease (AD). Strategically positioned at the brain-vasculature interface, PVMs sense the redox status, modulate immunity, and potentially influence ferroptosis-an iron-dependent form of regulated cell death increasingly implicated in AD. However, whether the involvement of PVMs in AD pathology specifically entails mechanisms related to the crosstalk between immunometabolism and ferroptosis, and the precise molecular pathways linking PVMs, immunometabolism, and ferroptosis to AD, remains unclear. We first obtained single-cell RNA sequencing data of PVMs from AD patients and control subjects via the GEO database, identified Differentially Expressed Genes (DEGs), and applied Mendelian Randomization (MR), with robustness validated via leave-one-out analysis to pinpoint key genes among the DEGs with causal relevance to AD. Next, we identified ferroptosis-related genes within these key genes and examined their associations with immune cell infiltration and immunometabolic signaling pathways, while also predicting their regulatory transcription factors to inform potential therapeutic strategies. We identified 149 DEGs in PVMs between AD and control groups, which were primarily enriched in immune and metabolic pathways. MR analysis established eight genes (ACSL1, SPATA6, RAB31, NIBAN1, HDAC4, GRAMD1B, GCC2, and DENND3) as causally and negatively associated with AD risk (IVW analysis identified all P < 0.05, with robustness confirmed by leave-one-out analysis), with ACSL1 being recognized as a known ferroptosis driver. Immune cell infiltration analysis revealed significant differences in monocyte and neutrophil proportions in AD, with DENND3 identified as the sole gene significantly correlated with monocyte abundance. The Key genes demonstrated distinct associations with immunometabolic pathways: GRAMD1B expression was positively associated with PI3K/AKT/mTOR signaling, whereas both NIBAN1 and SPATA6 showed enrichment in cells with high Notch signaling activity. ACSL1 exhibited robust associations with multiple pathways implicated in ferroptosis, including the IL-6/JAK/STAT3, interferon-γ, TGF-β, bile acid metabolism, and cholesterol homeostasis pathways, suggesting potential mechanisms that mediate the crosstalk between immunometabolism and ferroptosis. Transcription factor analysis highlighted shared regulation by CEBPD and the SP1/2/3/4 family, indicating convergent transcriptional control of these genes. This study identifies eight key genes in PVMs that may protect against AD through mechanisms involving the interplay between immunometabolism and ferroptosis. Our findings provide novel insights into the function of PVMs in AD pathophysiology and suggest potential therapeutic targets for this devastating neurodegenerative disease.

This experiment aimed to assess the effects of both preweaning nutrition and postweaning growth rate on the resilience of dairy heifers from birth to 20 months of age. Immune competence and metabolic characteristics were assessed via repeated vaccine immune challenges throughout early life. Heifers were subject to either a high or low preweaning nutritional treatment (high: 8 L vs. low: 4 L of milk per day). Calves in these treatment groups were then equally divided into either a high or low postweaning growth rate treatment until 20 months of age. Nutritional intake, growth and metabolic data can be found in a companion paper, while the current paper outlines the responses to the three immune challenges. In the preweaning phase, heifers on a high milk volume had superior immune competence, demonstrated by higher monocyte and eosinophil counts. All other immune biomarkers were not different between treatments. By 8 months of age, the differences in monocytes were lost; however, the differences in preweaning eosinophil counts remained at 8 months and through to 13 months of age. At 13 months of age, there were also three-way interaction effects of preweaning nutrition, postweaning growth rate and vaccination for white blood cell count and neutrophil count; however, the trends in these responses appear random and do not align towards any clear advantages of pre- or postweaning nutrition. Metabolic responses to the immune challenges do not suggest any form of carryover effect from the preweaning phase and seemed to reflect the nutritional input at the time.

BackgroundIdiopathic pulmonary fibrosis (IPF) is a degenerative respiratory condition characterized by significant mortality rates and a scarcity of available treatment alternatives. Cuproptosis, a novel form of copper-induced cell death, has garnered attention for its potential implications. The study aimed to explore the diagnostic value of cuproptosis-related hub genes in patients with IPF. Additionally, multiple bioinformatics analyses were employed to identify immune-related biomarkers associated with the diagnosis of IPF, offering valuable insights for future treatment strategies.MethodsFour microarray datasets were selected from the Gene Expression Omnibus (GEO) collection for screening. Differentially expressed genes (DEGs) associated with IPF were analyzed. Additionally, weighted gene coexpression network analysis (WGCNA) was employed to identify the DEGs most associated with IPF. Ultimately, we analyzed five cuproptosis-related hub genes and assessed their diagnostic value for IPF in both the training and validation sets. Additionally, four immune-related hub genes were screened using a protein–protein interaction (PPI) network and evaluated through the receiver operating characteristic (ROC) curve. Lastly, single-cell RNA-seq was employed to further investigate differential gene distribution.ResultsWe identified a total of 92 DEGs. Bioinformatics analysis highlighted five cuproptosis-related genes as candidate biomarkers, including three upregulated genes (CFH, STEAP1, and HDC) and two downregulated genes (NUDT16 and FMO5). The diagnostic accuracy of these five genes in the cohort was confirmed to be reliable. Additionally, we identified four immune-related hub genes that demonstrated strong diagnostic performance for IPF, with CXCL12 showing an AUROC of 0.90. We also examined the relationship between these four genes and immune cells. CXCL12 was significantly negatively associated with neutrophils, while CXCR2 was associated exclusively with neutrophils, consistent with our single-cell sequencing results. CTSG showed a primarily positive association with follicular helper T, and SPP1 was most strongly associated with macrophages. Finally, our single-cell sequencing data revealed that in patients with IPF, CXCL12 was highly expressed in the endothelial cell subset (ECs), while SPP1 exhibited high expression in multiple cellular populations. The expression of the CTSG showed statistically significant differences in monocyte macrophages.ConclusionThe research methodically depicted the intricate interplay among five cuproptosis-related genes, four immune-related hub genes, and IPF, offering new ideas for diagnosing and treating patients with IPF.

Post-traumatic osteoarthritis (PTOA) is a painful joint disease characterized by the degradation of bone, cartilage, and other connective tissues in the joint. PTOA is initiated by trauma to joint-stabilizing tissues, such as the anterior cruciate ligament, medial meniscus, or by intra-articular fractures. In humans, ~50% of joint injuries progress to PTOA, while the rest spontaneously resolve. To better understand molecular programs contributing to PTOA development or resolution, we examined injury-induced fluctuations in immune cell populations and transcriptional shifts by single-cell RNA sequencing of synovial joints in PTOA-susceptible C57BL/6J (B6) and PTOA-resistant MRL/MpJ (MRL) mice. We identified significant differences in monocyte and macrophage subpopulations between MRL and B6 joints. A potent myeloid-driven anti-inflammatory response was observed in MRL injured joints that significantly contrasted the pro-inflammatory signaling seen in B6 joints. Multiple CD206+ macrophage populations classically described as M2 were found enriched in MRL injured joints. These CD206+ macrophages also robustly expressed Trem2, a receptor involved in inflammation and myeloid cell activation. These data suggest that the PTOA resistant MRL mouse strain displays an enhanced capacity of clearing debris and apoptotic cells induced by inflammation after injury due to an increase in activated M2 macrophages within the synovial tissue and joint space.

Our study aimed to identify markers of enterococci's virulence potential by evaluating the properties of strains of different sites of isolation. Enterococcal strains were isolated as commensals from faeces and as invasive strains from the urine and blood of patients from the University Clinical Centre, Gdańsk, Poland. Changes in monocytes' susceptibility to the cytotoxic activity of isolates of different origins and their adherence to biofilm were evaluated using a flow cytometer. The bacterial protein profile was estimated by matrix assisted laser desorption ionization-time of flight mass spectrometer. The cytotoxicity of biofilm and monocytes' adherence to it were the most accurate factors in predicting the prevalence of the strain in the specific niche. Additionally, a bacterial protein with mass-to-charge ratio (m/z) 5000 was found to be responsible for the increased bacterial cytotoxicity, while monocytes' decreased adherence to biofilm was linked with the presence of proteins either with m/z 3330 or 2435. The results illustrate that monocytes' reaction when exposed to the bacterial biofilm can be used as an estimator of pathogens' virulence potential. The observed differences in monocytes' response are explainable by the bacterial proteins' profile. Additionally, the results indicate that the features of both bacteria and monocytes impact the outcome of the infection.

The pro-protein convertase FURIN (PCSK3) is implicated in a wide range of normal and pathological biological processes such as infectious diseases, cancer and cardiovascular diseases. Previously, we performed a systemic inhibition of FURIN in a mouse model of atherosclerosis and demonstrated significant plaque reduction and alterations in macrophage function. To understand the cellular mechanisms affected by FURIN inhibition in myeloid cells, we optimized a CRISPR-mediated gene deletion protocol for successfully deriving hemizygous (HZ) and nullizygous (NZ) FURIN knockout clones in U937 monocytic cells using lipotransfection-based procedures and a dual guide RNA delivery strategy. We observed differences in monocyte and macrophage functions involving phagocytosis, lipid accumulation, cell migration, inflammatory gene expression, cytokine release patterns, secreted proteomics (cytokines) and whole-genome transcriptomics between wild-type, HZ and NZ FURIN clones. These studies provide a mechanistic basis on the possible roles of myeloid cell FURIN in cardiovascular disorders.

Objective: The purpose of this study was to look at the seasonal distribution, age and gender distribution, and eosinophil, lymphocyte, and monocyte values according to age and gender in cases of insect bites that were brought to the emergency room over the course of a year. Materials and method: Retrospective analysis was performed on patients who were brought to the emergency room between 1.12.2021 and 1.12.2022 and had the ICD code W57 (Diagnosis Code - Bitten or stung by Nonvenomous Insects and Other Nonvenomous Arthropods). The following values were noted: age, gender, presenting season, CRP, Leukocyte, Platelet, Lymphocyte, Monocyte, Eosinophil, INR, PTZ, and Aptt levels. Findings: The study comprised a total of 694 patients—308 females and 386 males. The patients were 39.81 16.42 years old on average. Spring saw 9.4% of the patients, summer saw 67%, and fall saw 23.6%. According to the patients' gender, there were significant differences in the eosinophil (t:-3.535; p:0.0010.01) and monocyte (t:-4.909; p:0.0010.01) values. Regarding the season in which the patients were admitted, significant differences in lymphocyte (F:7.045; p:0.0010.01) and monocyte (F:3.208; p:0.0410.05) values were discovered. When the disparities in eosinophil, lymphocyte, and monocyte values were evaluated in relation to the patients' ages, significant differences in monocyte values were discovered (F:2.552; p:0.0270.05). Result: We commonly see insect bites in emergency rooms, which we can usually cure with straightforward remedies or occasionally without treatment, but in some unfortunate circumstances, we may have to deal with major issues and allergic responses (4). Almost little studies have been done on the seasonal distribution and evaluation of blood tests according to age and gender, despite the fact that there are many studies on this topic in the literature. We think that more study on this topic is necessary.

Dimethyl fumarate (DMF) is an immunomodulatory drug approved for the therapy of multiple sclerosis (MS). The identification of response biomarkers to DMF is a necessity in the clinical practice. With this aim, we studied the immunophenotypic and transcriptomic changes produced by DMF in peripheral blood mononuclear cells (PBMCs) and its association with clinical response. PBMCs were obtained from 22 RRMS patients at baseline and 12 months of DMF treatment. Lymphocyte and monocyte subsets, and gene expression were assessed by flow cytometry and next-generation RNA sequencing, respectively. Clinical response was evaluated using the composite measure "no evidence of disease activity" NEDA-3 or "evidence of disease activity" EDA-3 at 2 years, classifying patients into responders (n=15) or non-responders (n=7), respectively. In the whole cohort, DMF produced a decrease in effector (TEM) and central (TCM) memory T cells in both the CD4+ and CD8+ compartments, followed by an increase in CD4+ naïve T cells. Responder patients presented a greater decrease in TEM lymphocytes. In addition, responder patients showed an increase in NK cells and were resistant to the decrease in the intermediate monocytes shown by non-responders. Responder patients also presented differences in 3 subpopulations (NK bright, NK dim and CD8 TCM) at baseline and 4 subpopulations (intermediate monocytes, regulatory T cells, CD4 TCM and CD4 TEMRA) at 12 months. DMF induced a mild transcriptional effect, with only 328 differentially expressed genes (DEGs) after 12 months of treatment. The overall effect was a downregulation of pro-inflammatory genes, chemokines, and activators of the NF-kB pathway. At baseline, no DEGs were found between responders and non-responders. During DMF treatment a differential transcriptomic response was observed, with responders presenting a higher number of DEGs (902 genes) compared to non-responders (189 genes). Responder patients to DMF exhibit differences in monocyte and lymphocyte subpopulations and a distinguishable transcriptomic response compared to non-responders that should be further studied for the validation of biomarkers of treatment response to DMF.

Background: Periodontal diseases are inflammatory disorders caused by the accumulation of oral biofilm and the host response to this accumulation which characterized by exaggerated leukocytes and neutrophils attraction to the sites of inflammation by chemoattractants which are a very important part of the pathogenesis of periodontal diseases. This study aimed to determine and compare the clinical periodontal parameters and the leukocyte cell types in the peripheral blood between patients with gingivitis and periodontitis with different severities compared to healthy controls. Materials and methods: This study included 150 male subjects aged between 35-50 years. They were divided into three groups: gingivitis group (n=30), periodontitis patients (n=90) which subdivided into Mild =30 patients, Moderate =30 patients, Severe =30 patients and a control group (n=30) with clinically healthy periodontium. Clinical periodontal parameters were recorded ((plaque index (PLI), gingival index (GI), bleeding on probing (BOP), probing pocket depth (PPD) and clinical attachment level (CAL)). Blood samples were collected then an automated blood analyzer evaluated leukocyte cell types. Results: Significant differences in The counts of neutrophils and lymphocytes exhibited significant differences among the study groups and subgroups. On contrary, differences in monocytes, eosinophils, and basophils counts were not significant. Additionally, severity of periosontitis was significantly correlated with the mean counts of the various leukocyte cell types; however, clinical periodontal characteristics did not show such correlation with these inflammatory cells. Conclusion: This study demonstrated that periodontal disease with different severities is associated with possible episodes of bacteremia that originate from periodontal lesions which mediate inflammatory conditions that in turn causing changes in the systemic markers especially leukocytes cells types.

The present study was designed to detect possible biomarkers associated with Type 1 diabetes mellitus (T1DM) incidence in an effort to develop novel treatments for this condition. Three mRNA expression datasets of peripheral blood mononuclear cells (PBMCs) were obtained from the GEO database. Differentially expressed genes (DEGs) between T1DM patients and healthy controls were identified by Limma package in R, and using the DEGs to conduct GO and DO pathway enrichment. The LASSO-SVM were used to screen the hub genes. We performed immune correlation analysis of hub genes and established a T1DM prognosis model. CIBERSORT algorithm was used to identify the different immune cells in distribution between T1DM and normal samples. The correlation of the hub genes and immune cells was analyzed by Spearman. ROC curves were used to assess the diagnostic value of genes in T1DM. A total of 60 immune related DEGs were obtained from the T1DM and normal samples. Then, DEGs were further screened to obtain 3 hub genes, ANP32A-IT1, ESCO2 and NBPF1. CIBERSORT analysis revealed the percentage of immune cells in each sample, indicating that there was significant difference in monocytes, T cells CD8+, gamma delta T cells, naive CD4+ T cells and activated memory CD4+ T cells between T1DM and normal samples. The area under curve (AUC) of ESCO2, ANP32A-IT1 and NBPF1 were all greater than 0.8, indicating that these three genes have high diagnostic value for T1DM. Together, the findings of these bioinformatics analyses thus identified key hub genes associated with T1DM development.

Background. After trauma, the subtypes of white blood cells (WBCs) in circulation and the derived neutrophil-to-lymphocyte ratio (NLR), monocyte-to-lymphocyte ratio (MLR), and platelet-to-lymphocyte ratio (PLR) may undergo relative changes and reflect the patients’ immune-inflammatory status and outcome. This retrospective study was designed to investigate the relationship between these variables and the mortality outcomes in adult patients with polytrauma, which is defined as an abbreviated injury scale (AIS) score ≥ 3 in two or more different body regions. Methods. A comparison of the expression of subtypes of WBCs, NLR, MLR, and PLR upon arrival to the emergency department was performed in selected propensity score-matched patient cohorts created from 479 adult patients with polytrauma between 1 January 2015 and 31 December 2019. A multivariate logistic regression analysis was used to identify the independent risk factors for mortality. Results. There were no significant differences in monocyte, neutrophil, and platelet counts, as well as in MLR, NLR, and PLR, between deceased (n = 118) and surviving (n = 361) patients. In the propensity score-matched patient cohorts, which showed no significant differences in sex, age, comorbidities, and injury severity, deceased patients had significantly higher lymphocyte counts than survivors (2214 ± 1372 vs. 1807 ± 1162 [106/L], respectively, p = 0.036). In addition, the multivariate logistic regression analysis revealed that the lymphocyte count (OR, 1.0; 95% confidence interval [CI], 1.00–1.06; p = 0.043) was a significant independent risk factor for mortality in these patients. Conclusions. This study revealed that there was no significant difference in the counts of monocytes, neutrophils, and platelets, as well as in MLR, NLR, and PLR, between deceased and surviving patients with polytrauma. However, a significantly higher lymphocyte count may be associated with a worse mortality.

BackgroundMembranous nephropathy (MN) is a common pathological phenotype for adult nephrotic syndrome (NS). The occurrence of MN is increasing across China, but diagnostic methods for MN still rely on kidney biopsy and PLA2R and THSD7A detection in plasma and kidney tissue, and there has been no new biomarker for MN discovered since 2014. Immune infiltration status in MN patients suffers from the dearth of associated studies. In the present study, we aimed to find new bio-markers for MN and evaluate the role of immune cells infiltration in MN pathology.MethodsWe downloaded MN expression profile from the Gene Expression Omnibus database and used R-project to screen differentially expressed genes (DEGs) and performed functional correlation analysis. Least absolute shrinkage and selection operator (LASSO) logistic regression and Radom Forest algorithms were used to screen and verify the bio-markers of MN. Finally, CIBERSORT was used to evaluate the infiltration of immune cells in MN tissues.ResultsA total of 463 DEGs were screened from the MN tissue in this study. ETS2 was identified as bio-marker for MN. The CIBERSORT results showed that there were statistical differences in monocytes, plasma cells, regulatory T cells, and memory B cells. In addition, ETS2 was positively related to monocytes, M1 phase macrophages, and neutrophils and negatively correlated to plasma cells, CD4+ T memory cells, M2 macrophages, CD8+ T cells, memory B cells, and resting mast cells.Conclusion(1) Machine learning algorithms reveals Ets2 as a novel target for membranous nephropathy patients. (2) Immune infiltration plays an important part in membranous nephropathy. (3) Ets2 expression is related to immune cells infiltration.

Background: Carotid artery disease accounts for 30% of ischemic strokes in the general population. Numerous biomarkers have been investigated for predicting either the progression or the severity of the disease. The aim of this retrospective study was to compare hematologic indices among patients referred for surgical interventions due to severe carotid disease. Methods: In total, 135 patients (87 (64.4%) men and 48 (35.6%) women) with a mean age of 70 ± 8 years who underwent surgical carotid intervention were enrolled into the study. Results: A Mann–Whitney test for independent samples revealed significant differences in monocyte to lymphocyte ratio (MLR) and mean corpuscular hemoglobin concentration (MCHC) between patients with one and two (collateral) carotid diseases. The cut-off value for MLR was 0.3 (AUC = 0.654, p = 0.048, 70.0% sensitivity and 74.6% specificity) and for MHCH was 21.6. (AUC = 0.730, p < 0.001, 70.0% sensitivity and 77.2% specificity). A multivariable model of logistic regression revealed two significant parameters for collateral carotid stenosis disease including MLR > 0.3 (OR 6.19 with 95% CI 2.02–19.01, p = 0.001) and MCHC > 21.6 (OR 7.76, 95% CI 2.54–23.72, p < 0.001). Conclusions: MLR above 0.3 and MCHC above 21.6 have predictive values for colleterial carotid stenosis and may be used as easily accessible indicators for atherosclerosis severity.

Abstract Background and aim Cardiometabolic risk accrues across the entire life course and childhood is a key epoch for effective prevention. Obesity in childhood is the most prevalent modifiable risk factor for later cardiovascular disease (CVD). Inflammatory biomarkers and innate immune capacity are increased in adults with obesity, but childhood data are scarce. We aimed to investigate (i) innate immune cell activation in children with and without obesity; and (ii) whether weight loss impacts the innate immune inflammatory phenotype. Methods The innate immune phenotype of Peripheral Blood Mononuclear Cells (PBMCs) from 31 children with obesity (BMI z-score&amp;gt;2.5) and 22 children of healthy weight (−1.5≤BMIz≤1.5, sex, age and pubertal stage matched) was characterized by high dimensional flow cytometry, ex vivo stimulation assays with subsequent 27-plex cytokine measurements, and transcriptome analysis using RNA sequencing (Figure 1). Children with obesity participated to the Royal Children's Hospital Weight Management Service (median 5 years) and at follow-up, PBMCs were obtained again as well as anthropometric data and subclinical cardiovascular phenotypes. Results Flow cytometric analysis showed marked differences in cell composition between children with obesity and children of healthy weight. Specifically, children with obesity have significant changes in monocyte subsets and an increased expression of monocyte activation markers. Upon stimulation, monocytes of children with obesity show an increased cytokine production capacity. Finally, transcriptomic analysis shows significant differences between monocytes from obese children and healthy controls. Effects of weight loss on these immune parameters and correlations with preclinical CVD phenotypes are currently being analysed. Conclusions Monocytes from children with obesity have a pro-inflammatory phenotype compared to children of normal weight. Heightened inflammation may contribute to increased CVD risk later in life and may offer opportunities for early intervention. Funding Acknowledgement Type of funding sources: Other. Main funding source(s): Dutch Scientific Organisation (NWO) - Rubicon grant to S.B. Dutch Heart Foundation - CVON IN CONTROL II to N.P.R. and D.B. Figure 1. Schematical overview of study

Functional and transcriptional differences in monocytes from children with obesity compared to children of healthy weight

Background and AimsPatients with chronic kidney disease (CKD) are at higher risk of severe complications and mortality due to Covid-19 than patients with other known risk factors. The association between CKD and mortality persist in analyses adjusted for covariates known to associate with worse Covid-19 outcomes, suggesting that CKD confers a risk beyond that associated to comorbid conditions. The mechanisms underlying the increased susceptibility to severe Covid-19 in CKD remains unclear but morphologic and functional differences in monocytes have been associated with prolonged hospitalization in other cohorts of patients. The monocyte is capable to contribute to the pathophysiology through different mechanisms. There is however insufficient information on factors that orchestrate different aspects of monocyte function and how these factors relate to outcomes. Increased knowledge into which features of monocyte function that contribute to risks in CKD patients with Covid-19 is important to guide treatment strategies. The aim of the present study was to examine the concentrations of monocyte chemoattractant markers MCP-1 (Monocyte Chemoattractant Protein-1; CCL2) and MIP-1α (Macrophage Inflammatory Protein 1-α; CCL3) in patients with Covid-19 and normal or impaired kidney function and to compare that to CKD patients matched for sex and eGFR, and sex matched healthy subjects. We analyzed the impact of these monocyte chemoattractant markers on in-hospital and 30 days mortality by logistic and multiple regression analyses. We related this to established risk factors for morbidity and mortality in Covid-19 patients, e.g. CRP and IL-6.MethodWe prospectively included 110 patients with Covid‐19 (mean age 59 yr., mean eGFR 75 ml/min/1.73m2) admitted to Danderyd University Hospital, Stockholm, Sweden, during the first pandemic wave in April to May 2020 and 33 sex and eGFR-matched patients (mean age 51 yr., eGFR 52 ml/min/1.73m2) with CKD and 35 sex matched healthy subjects (mean age 47 yr., eGFR 101 ml/min/1.73m2).We used Luminex assays to analyze MCP-1, MIP-1α and IL-6 and routine laboratory tests to determine white blood cell count (WBC) and CRP.ResultsPatients with Covid-19 had significantly lower concentrations of MIP-1α (p<0.001), and higher IL-6 (p<0.001) and CRP (p<0.001) than patients with CKD and healthy subjects (Kruskal-Wallis), there were no differences in MCP-1 between groups. We found significant negative correlations between MCP-1 (p<0.05), MIP-1α (p<0.05) and IL-6 (p<0.05) with eGFR in patients with Covid-19 (Spearman´s rank correlation). Logistic regression analysis (Odds ratio, OR, 95% Confidence Intervals (CI)) and Cox proportional hazard models (Hazard ratio, HR), both adjusted for age, showed significant associations between in-hospital mortality and WBC, CRP, IL-6, MCP-1 and MIP-1α (Table, Figure). Similar findings were observed also for 30 days mortality.MO132 Table.Logistic regression analysis, odds ratio (OR) and Cox proportional hazard ratio (HR) both adjusted for age with 95% CI of in-hospital mortality in patients with Covid-19Logistic regression analysisCox proportional hazard analysisp-valueOR95% CIp-valueHR95% CILowerUpperLowerUpperWhite BC =0.001 1.4381.1671.771 <0.001 1.2511.1281.387CRP =0.01 1.0081.0021.015 0.004 1.0081.0031.013IL-6 =0.05 1.0071.0001.013 0.001 1.0011.0011.002MCP-1 <0.05 1.0011.0001.002 0.001 1.0001.0001.001MIP-1α =0.006 1.0061.0021.011 0.001 1.0051.0021.008ConclusionWe demonstrate that factors related to monocyte recruitment and activation, MCP-1 and MIP-1α, are associated with in-hospital and 30-days mortality in CKD patients with Covid-19. In this patient group general inflammatory markers as IL-6 and CRP are also associated with risk of mortality. These data contribute to an increased understanding of the impact of monocyte activation in Covid-19 and may be of value when treatment strategies are evaluated.