Gene Expression In Mononuclear Cells Research Articles

Analysis of Peripheral Blood Mononuclear Cells Gene Expression Highlights the Role of Extracellular Vesicles in the Immune Response following Hematopoietic Stem Cell Transplantation in Children

Hematopoietic stem cell transplantation (HSCT) is an effective treatment method used in many neoplastic and non-neoplastic diseases that affect the bone marrow, blood cells, and immune system. The procedure is associated with a risk of adverse events, mostly related to the immune response after transplantation. The aim of our research was to identify genes, processes and cellular entities involved in the variety of changes occurring after allogeneic HSCT in children by performing a whole genome expression assessment together with pathway enrichment analysis. We conducted a prospective study of 27 patients (aged 1.5–18 years) qualified for allogenic HSCT. Blood samples were obtained before HSCT and 6 months after the procedure. Microarrays were used to analyze gene expressions in peripheral blood mononuclear cells. This was followed by Gene Ontology (GO) functional enrichment analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, and protein–protein interaction (PPI) analysis using bioinformatic tools. We found 139 differentially expressed genes (DEGs) of which 91 were upregulated and 48 were downregulated. “Blood microparticle”, “extracellular exosome”, “B-cell receptor signaling pathway”, “complement activation” and “antigen binding” were among GO terms found to be significantly enriched. The PPI analysis identified 16 hub genes. Our results provide insight into a broad spectrum of epigenetic changes that occur after HSCT. In particular, they further highlight the importance of extracellular vesicles (exosomes and microparticles) in the post-HSCT immune response.

Changes in immune system and intestinal bacteria of cows during the transition period

High-yield dairy cows need high energy feed during periods of increased milk production. The transitional feeding to high energy feed increases the risk of developing a variety of metabolic disorders. Here, five Holstein cows were fed a four-stage feeding protocol (3 weeks for each stage) ranging from 54.9 to 73.7% total digestive nutrients (TDN). The purpose of the study was to investigate the effect of lactic acid bacteria on high-energy-fed cows associated with transitional feeding, and to evaluate the effects of probiotics on intestinal bacterial changes and inflammatory responses. Three feed transition periods were established for five cows, and Lactobacillus plantarum RGU-LP1 (LP1) was fed as a probiotic during the high-energy feeding period. The number of lymphocyte subsets such as CD3-, CD4-, and CD8 positive cells decreased in response to the high energy feed. Lipopolysaccharide (LPS)-induced cytokine (IL-1β and IL-2) gene expression in peripheral blood mononuclear cells (PBMCs) was shown to increase in those animals receiving the high energy feed. However, supplementation with LP1 resulted in an increase in the number of lymphocyte subsets and the expression of IL-1β and IL-2 were returned to the level at low energy diet. These results suggest that high energy diets induce inflammatory cytokine responses following LPS stimulation, and that the addition of LP1 mitigates these results by regulating the LPS-induced inflammatory reaction. Therefore, the functional lactic acid bacteria LP1 is expected to regulate inflammation resulting from high energy feeding, and this probiotic could be applied to support inflammatory regulation in high-yield dairy cows.

Involvement of lncRNA IL21-AS1 in interleukin-2 and T follicular regulatory cell activation in systemic lupus erythematosus

BackgroundThe single nucleotide polymorphism (SNP) rs62324212, located in IL21 antisense RNA 1 (IL21-AS1), has been identified as a genetic risk variant associated with systemic lupus erythematosus (SLE). We aimed to probe the characteristics of IL21-AS1 and explore its clinical relevance focusing on T helper subsets and disease activity in patients with SLE.Methodsrs62324212 genotyping was determined using allelic discrimination by quantitative PCR. Gene expression in peripheral blood mononuclear cells and cell surface markers in CD4+ T cells were analyzed using PCR and flow cytometry. The association among IL21-AS1, CD4+ T cell subsets, and SLE disease activity was accessed.ResultsEnsembl Genome Browser analysis revealed that rs62324212 (C>A) was located in the predicting enhancer region of IL21-AS1. IL21-AS1 was expressed in the nucleus of CD4+ T and B cells, but its expression was decreased in patients with SLE. IL21-AS1 expression was positively correlated with mRNA levels of IL-2 but not IL-21, and it was associated with the proportion of activated T follicular regulatory (Tfr) cells. Furthermore, we observed a significant negative correlation between IL21-AS1 expression and disease activity in patients with SLE (n = 53, p < 0.05).ConclusionIL21-AS1 has an effect on disease activity through an involvement of IL-2-mediated activation of Tfr cells in SLE. Thus, targeting the IL21-AS1 may provide therapeutic approaches for SLE.

Replacing Saturated Fat with Polyunsaturated Fat Modulates Peripheral Blood Mononuclear Cell Gene Expression and Pathways Related to Cardiovascular Disease Risk Using a Whole Transcriptome Approach.

The aim of this study is to explore the molecular mechanisms underlying the effect of replacing dietary saturated fat (SFA) with polyunsaturated fat (PUFA) on cardiovascular disease (CVD) risk using a whole transcriptome approach. Healthy subjects with moderate hypercholesterolemia (n = 115) are randomly assigned to a control diet (C-diet) group or an experimental diet (Ex-diet) group receiving comparable food items with different fatty acid composition for 8 weeks. RNA isolated from peripheral blood mononuclear cells (PBMCs) at baseline and after 8 weeks of intervention is analyzed by microarray technology (n = 95). By use of a linear regression model (n = 92), 14 gene transcripts are differentially altered in the Ex-diet group compared to the C-diet group. These include transcripts related to vascular smooth muscle cell proliferation, low-density lipoprotein receptor folding, and regulation of blood pressure. Furthermore, pathways mainly related to immune response and inflammation, signal transduction, development, and cytoskeleton remodeling, gene expression and protein function, are differentially enriched between the groups. Replacing dietary SFA with PUFA for 8 weeks modulates PBMC gene expression and pathways related to CVD risk in healthy subjects with moderate hypercholesterolemia.

Gene expression analysis of MAOA and the clock gene ARNTL in individuals with bipolar disorder compared to healthy controls

Introduction Circadian rhythms are associated with bipolar disorder (BD). This cross-sectional study aimed at investigating ARNTL and MAOA gene expression differences (1) between individuals with BD and controls, (2) between affective episodes, and (3) the relationship between ARNTL and MAOA expression. Methods ARNTL and MAOA gene expression in peripheral mononuclear blood cells were analysed from fasting blood samples (BD n = 81, controls n = 54) with quantitative real-time PCR operating on TaqMan® assays (normalised to 18S RNA expression). ANCOVAs corrected for age, sex, body mass index, and medication was used to evaluate expression differences and correlation analyses for the relation between ARNTL and MAOA expression. Results ARNTL gene expression differed between affective episodes (F(2,78) = 3.198, p = 0.047, Partial Eta2= 0.083), but not between BD and controls (n.s.). ARNTL and MAOA expression correlated positively in BD (r = 0.704, p < 0.001) and in controls (r = 0.932, p < 0.001). MAOA expression differed neither between BD and controls nor between affective episodes (n.s.). Discussion Clock gene expression changes were observed in different affective states of BD. More precisely, ARNTL gene expression was significantly higher in euthymia than in depression. ARNTL and MAOA gene expression correlated significantly in BD and in controls, which emphasises the strong concatenation between circadian rhythms and neurotransmitter breakdown.

Gene expression in peripheral blood mononuclear cells in impulsive aggression: Intermittent explosive disorder compared with non-aggressive healthy and psychiatric controls

Evidence of chronic, systemic, low levels of inflammation is present in several stress-related psychiatric conditions including schizophrenia, depression, and intermittent explosive disorder (IED). We analyzed leukocyte gene expression (mRNA) to quantify the activity of pro and anti-inflammatory signaling pathways. Work performed in non-aggressive populations has uncovered a Conserved Transcriptional Response to Adversity (CTRA) characterized by an upregulation of pro-inflammatory gene transcription in chronically stressed individuals. We used pathway-based bioinformatic analyses of genome-wide transcriptional profiles of peripheral blood leukocyte samples from IED study participants (N = 45) and controls [healthy (n = 45) and psychiatric (n = 34)], with analyses focusing on the pro-inflammatory transcription control pathway mediated by the NF-kB family of transcription factors (typically upregulated in CTRA) and the antiviral transcription control pathway mediated by anti-viral response (IRF) family transcription factors (typically downregulated in CTRA). Compared with both healthy and psychiatric controls, individuals with IED had upregulated transcriptional activity of the antiviral response (IRF), but no evidence of pro-inflammatory NF-kB activation. Analyses implicated CD4 + T cells, CD8 + T cells, and B lymphocytes in IED-related transcriptional alterations, but showed no significant indication of monocyte involvement. This suggests that the inflammatory profile of IED differs substantially from that observed previously in other stress-related disorders, and may involve a pathogen-driven adaptive immune etiology.

Investigating the Correlation between Growth Differentiation Factor 15 Serum Level and Its Gene Expression with Psoriasis and Its Severity.

Psoriasis is a chronic inflammatory dermatitis characterized by an inflammatory epidermal hyperproliferation. Growth differentiation factor-15 (GDF-15), a member of the transforming growth factor-β family, has immune modulatory roles in autoimmune condition of Psoriasis. This study aimed to evaluate the relationship between GDF-15 serum levels as well as gene expression with psoriasis and its severity. This case-control study was performed on 45 patients with psoriasis Vulgaris and 45 healthy individuals. The severity of the disease was determined based on the psoriasis area and severity index (PASI score). Serum levels of GDF-15 were measured by enzyme-linked immunosorbent assay (ELISA) and its gene expression in peripheral blood mononuclear cells was quantified by real-time polymerase chain reaction (RT-PCR). The mean serum levels of GDF-15 in patients and controls were 1.98±1.57 ng/mL and 0.93±0.48 ng/mL, respectively. GDF-15 gene expression was measured as 9.7±6.6% in the patient group and 7.6±2.5% in the healthy group. The mean of GDF-15 serum levels in mild, moderate, and severe cases of psoriasis were 0.45±0.35, 2.27±0.7, and 3.5±1.6 ng/mL, respectively, indicating that elevated serum levels of GDF-15 correlate significantly with disease severity. The mean of GDF-15 gene expression in the mild, moderate, and severe forms of psoriasis were 5.25±3.2, 7.6±2.8, and 17.8±5.7, respectively which indicate a significant relationship between GDF-15 gene expression and psoriasis severity. Based on this study, in psoriatic patients, GDF-15 serum levels and gene expression are significantly higher than those in healthy controls. Such values were correlated with disease activity, especially in severe cases. Therefore, GDF-15 may be used as a prognostic marker of psoriasis.

Immunomodulatory Effect of Propolis on Foxp3 Gene Expression in Human Peripheral Blood Mononuclear Cells Stimulated in vitro with Pseudomonas Aeruginosa Ag.

Immune balance during infection is critical for both supporting the defense of the immune system of the body and preventing an overly aggressive immune response. Foxp3, a transcription factor of regulatory T cells, plays a critical role in balancing the immune system of the body. Propolis has been shown to affect Foxp3 expression. This study aimed to verify the effect of propolis extracts on in vitro Foxp3 gene expression in peripheral blood mononuclear cells (PBMCs) stimulated with Pseudomonas aeruginosa Ag. In this study, a total of 20 apparently healthy volunteers were included, with 10 males and 10 females within the age range of 20-40 years old. Five ml of blood were drawn from each participant to assess Foxp3 gene expression in PBMCs using density gradient lymphoprep and stimulated with P.aeruginosa lipopolysaccharide (LPS) in vitro. The samples were divided into four distinct groups as follows: LPS stimulated PBMCs, ethanol-extracted propolis (EEP) + LPS stimulated PBMCs, and water-extracted propolis (WEP) + LPS stimulated PBMCs and PBMCs as the control group. The Foxp3 gene expression level was estimated in all four groups following a period of 48 h of cultivation by real-time polymerase chain reaction technique using SYBR green dye. Results of the study indicated that propolis had a great effect on the mRNA Foxp3 expression. Both EEP and WEP had immunomodulatory effects through the Foxp3 mRNA expression, both the EEP and WEP could significantly inhibit Foxp3 mRNA gene expression by human PBMCs after stimulation with pseudomonas Ag in vitro. Propolis exhibited an immunoregulatory effect which was the same with ethanol and water extracts on Foxp3 mRNA gene expression.

Effect of liraglutide on expression of inflammatory genes in type 2 diabetes

Anti-inflammatory effects of glucagon-like peptide 1 receptor agonist (GLP-1 RA) treatment in T2D may contribute to the cardiovascular benefits observed with GLP-1 RAs in outcome studies. We investigated if the GLP-1 RA liraglutide exerts anti-inflammatory effects through modulation of inflammatory gene expression in peripheral blood mononuclear cells (PBMCs). From 54 participants of a double-blinded trial where individuals with type 2 diabetes (T2D) were randomized to liraglutide (1.8 mg/day) or placebo for 26 weeks, a sub-study was performed in which PBMCs were extracted from fresh blood at study start and at end-of-treatment. The expression of selected inflammatory genes in PBMCs were measured by quantitative real-time polymerase chain reaction (PCR). Moreover, the expression of the GLP-1 receptor (GLP1R) was examined in a subset (n = 40) of the PBMC samples. The human monocytic cell line THP-1 was used for in vitro GLP-1 exposure experiments. The expression of tumor necrosis factor-α (TNFA) (p = 0.004) and interleukin-1β (IL1B) was downregulated (p = 0.046) in the liraglutide-treated group (n = 31), and unchanged in the placebo group (n = 21, p ≥ 0.11), with no significant differences between the two groups (p ≥ 0.67). The expression of interferon-γ (IFNG) and cluster of differentiation 163 (CD163) were upregulated in both groups (p ≤ 0.006) with no differences between groups (p ≥ 0.47). C–C Motif Chemokine Ligand 5 (CCL5) was upregulated in the liraglutide-treated group (p = 0.002) and unchanged in the placebo group (p = 0.14), with no significant difference between groups (p = 0.36). Intercellular adhesion molecule 1 (ICAM1) was unchanged in both groups (p ≥ 0.43). GLP1R expression in the PBMCs was undetectable. In vitro experiments showed no effect of GLP-1 treatment on inflammatory gene expression in THP-1 cells. GLP1R expression in THP-1 cells was not detectable. In summary, we observed a discrete modulatory effect of liraglutide on the expression of inflammatory genes in PBMCs. The lack of evidence for GLP1R expression in PBMCs and THP-1 cells suggests that possible effects of liraglutide on the PBMCs’ gene expression are most likely indirect. Further investigations are needed to establish the anti-inflammatory potential of GLP-1 RAs.

Abstract P139: Diet-induced Shifts In HDL Profiles Are Associated With Changes In Immune Parameters

Introduction: Preclinical and population-based studies have demonstrated immunomodulatory and anti-inflammatory properties of HDL; however, it is unclear whether diet-induced changes in HDL profiles can directly impact markers of immune inflammation. Hypothesis: We hypothesized that daily consumption of different egg-based diets, which have been shown to differentially impact serum lipids and lipoprotein profiles, would induce changes in HDL parameters that correspond to shifts in clinical and molecular immune markers. Methods: Healthy men and women (18-35y, BMI &lt; 30kg/m 2 or &lt; 30% body fat for men and &lt; 40% body fat for women, n = 26) participated in a 16-week randomized, crossover intervention trial (NCT03577223), in which they consumed an egg-free diet for 4 weeks, followed by a 4-week diet containing either 3 whole eggs or 3 egg whites per day. Participants then followed a 4-week egg-free diet washout period, before switching to the alternative whole egg or egg white diet treatment. Fasting lipoprotein profiles, complete blood cell counts, and gene expression in peripheral blood mononuclear cells (PBMCs) was measured at the end of each diet period. Results: While no changes in total HDL particles, average HDL particle diameter, or the concentration of small (7.4-8.0 nm) or medium (8.1-9.5 nm) HDL particles was observed, the concentration of large HDL particles (10.8nm) was increased by whole egg intake as compared to intake of egg whites. A trend toward increased apolipoprotein A1 during the whole egg diet period as compared to the egg-free diet period was additionally observed. In line with our hypothesis, changes in large HDL concentrations between whole egg vs. egg white periods were inversely correlated with total white blood cell counts and absolute lymphocyte counts. CD8A mRNA expression in PBMCs was inversely correlated with concentrations of total and large HDL particles. Importantly, while serum choline and betaine were increased by whole egg intake compared to the other diet periods, no differences in pro-atherogenic and inflammatory trimethylamine-N-oxide (TMAO) between diet periods was observed. Conclusion: These findings suggest that diet-induced changes in HDL profiles are directly associated with changes in immune parameters.

Marked up-regulation of anti inflammatory cytokine gene expression in the peripheral blood mononuclear cells of postpartum cows with endometritis

This study was designed to determine the mRNA expression profile of inflammatory and anti-inflammatory cytokines in peripheral blood mononuclear cells (PBMC) of endometritic and non-endometritic cows to find out a possible marker for diagnosis of endometritis. Cows (21) including 12 endometritic, and 9 normal (non-endometritic) that did not develop uterine diseases postpartum (pp) were selected. Expression profile (mRNA) of cytokines, viz. IL-1β, TNFα, IL-8 and IL-4 in endometritic cows relative to non-endometritic was studied in PBMC isolated from the blood of the cows collected within 4 to 5 weeks pp. Fold difference (n-fold) in cytokine gene expression in the PBMC of endometritic cows relative to normal was calculated using relative quantification method (2-ΔΔCt). The relative fold change in gene expression for IL-1 was significantly lower in endometritic animals (0.31) than in cows that did not develop endometritis. Significant up-regulation was noted for the proinflammatory cytokine TNFα (9.53 fold and anti-inflammatory cytokine IL-4 (90.09) in cows suffering from endometritis relative to nonendometritic cows. Up-regulation of chemokine IL-8 gene (6.25 fold; was also observed in endometritic compared to non-endometritic cows. Highest and significant up-regulation of mRNA expression for IL-4 followed by TNFα in PBMC of endometritic cows recorded in the study may indicate the development of endometritis in postpartum cows.

MYO1F regulates antifungal immunity by regulating acetylation of microtubules

Opportunistic fungal infections have become one of the leading causes of death among immunocompromised patients, resulting in an estimated 1.5 million deaths each year worldwide. The molecular mechanisms that promote host defense against fungal infections remain elusive. Here, we find that Myosin IF (MYO1F), an unconventional myosin, promotes the expression of genes that are critical for antifungal innate immune signaling and proinflammatory responses. Mechanistically, MYO1F is required for dectin-induced α-tubulin acetylation, acting as an adaptor that recruits both the adaptor AP2A1 and α-tubulin N-acetyltransferase 1 to α-tubulin; in turn, these events control the membrane-to-cytoplasm trafficking of spleen tyrosine kinase and caspase recruitment domain-containing protein 9 Myo1f-deficient mice are more susceptible than their wild-type counterparts to the lethal sequelae of systemic infection with Candida albicans Notably, administration of Sirt2 deacetylase inhibitors, namely AGK2, AK-1, or AK-7, significantly increases the dectin-induced expression of proinflammatory genes in mouse bone marrow-derived macrophages and microglia, thereby protecting mice from both systemic and central nervous system C. albicans infections. AGK2 also promotes proinflammatory gene expression in human peripheral blood mononuclear cells after Dectin stimulation. Taken together, our findings describe a key role for MYO1F in promoting antifungal immunity by regulating the acetylation of α-tubulin and microtubules, and our findings suggest that Sirt2 deacetylase inhibitors may be developed as potential drugs for the treatment of fungal infections.

Chaya Leaf Decreased Triglycerides and Improved Oxidative Stress in Subjects With Dyslipidemia.

Chaya is an edible leaf popular in Mexico and Central America because of its high nutritional value. Studies in animal models have demonstrated the beneficial effects of Chaya, which include reduction of circulating lipids and increase in antioxidant activity. However, its hypolipidemic and antioxidant effects have not been demonstrated in humans. Thus, the aim of the present study was to evaluate the effect of Chaya on the lipid profile, lipid peroxidation, inflammation, and peripheral blood mononuclear cell gene expression in a population with dyslipidemia. We performed a single-arm trial in 30 participants with dyslipidemia who consumed 500 mL of Chaya beverage per day over a 6-week period. Interestingly, we observed a significant decrease in serum triglyceride concentration (P < 0.05) and an increase in plasma antioxidant activity and polyphenol concentration (P < 0.005) after 6 weeks of Chaya consumption. This was accompanied by a reduction in the oxidative stress marker MDA (P < 0.0001) and by an increase in the antioxidant enzyme CAT expression in peripheral blood mononuclear cells (P < 0.001). Altogether, our results demonstrate that consumption of Chaya has hypotriglyceridemic and antioxidant effects in subjects with dyslipidemia.

Effect of lysed and non-lysed sickle red cells on the activation of NLRP3 inflammasome and LTB4 production by mononuclear cells.

This study tested the hypothesis that sickle red blood cell (SS-RBC) can induce inflammasome NLRP3 components gene expression in peripheral blood mononuclear cells (PBMCs) as well as interleukin-1β (IL-1β) and leukotriene B4 (LTB4) production. Additionally, we investigated the effect of hydroxyurea (HU) treatment in these inflammatory markers. PBMCs from healthy donors (AA-PBMC) were challenged with intact and lysed RBCs from SCA patients (SS-RBC) and from healthy volunteers (AA-RBC). NLRP3, IL-1β, IL-18 and Caspase-1 gene expression levels were assessed by quantitative PCR (qPCR). IL-1β protein levels and LTB4 were measured by ELISA. We observed that lysed SS-RBC induced the expression of inflammasome NLRP3 components, but this increase was more prominent for CASP1 and IL18 expression levels. Moreover, we observed that intact SS-RBC induced higher production of IL-1β and LTB4 than lysed SS-RBC. Although SCA patients treated with HU have a reduction in NLRP3 gene expression and LTB4 production, this treatment did not modulate the expression of other inflammasome components or IL-1β production. Thus, our data suggest that caspase-1, IL-1β and IL-18 may contribute to the inflammatory status observed in SCA and that HU treatment may not interfere in this inflammatory pathway.

Reduced Hemolysis and Related Complications in Females with Sickle Cell Disease

Sickle cell disease (SCD) is an inherited erythrocyte disorder characterized by chronic hemolysis and vasculopathy. Females have reduced levels of hemolysis compared to males in both the general population (Jordan A et al, Vox Sang 2016) as well as in SCD (Kanias T et al, Transfusion 2016). This phenomenon may be due to the effects of gender on hemoglobin (Hb) F levels (Dover GJ et al, Blood 1992), red blood cell membrane stability (Jordan A et al, Vox Sang 2016), and nitric oxide (NO) bioavailability (Gladwin MT et al, Circulation 2003). We wished to confirm reduced hemolysis in SCD females and whether this would be associated with reduced hemolytic complications, some of which are associated with mortality. Finally, we wished to see if there were suggestions of improved survival in females compared to males with SCD.We investigated differences in laboratory and clinical features by gender in 395 (155 male, 240 female) SCD patients treated at the University of Illinois at Chicago (UIC) and in 668 (304 male, 364 female) SCD patients from the multicenter Walk-PHaSST cohort. Comparisons of linear and categorical variables were performed using ANOVA and logistic regression analysis, respectively adjusting for age, SCD genotype, and Hb F levels. Median values and interquartile range (IQR) are provided.In SCD patients either not on hydroxyurea or with Hb F levels available prior to initiating hydroxyurea, we observed higher Hb F values in females vs. males from both the UIC (n=329, P=0.004) and Walk-PhaSST (n=368, P=0.003) cohorts (Table 1). Hemolytic markers were lower in females vs. males and these relationships persisted after adjusting for HbF levels, including LDH (UIC: P=0.0004, Walk-PHaSST: P=0.01); total bilirubin (UIC: P=0.008; Walk-PHaSST: P=0.002), AST (UIC: P=0.08; Walk-PHaSST: P=0.0002), and absolute reticulocyte count (UIC: P=0.04, Walk-PHaSST: P=0.01) (Table 1).Less hemolysis would be expected to decrease the occurrence or severity of a hemolytic vasculopathy and its manifestations. Pulse pressures were lower in females than males in both cohorts (UIC: P <0.0001; Walk-PHaSST: P=0.0003). Additional manifestations less frequently observed in females than males included leg ulcer history (UIC: P<0.0001; Walk-PHaSST: P=0.09) and tricuspid regurgitant velocity >3.0m/s (UIC: P=0.2; Walk-PHaSST: P=0.01)(Table 1). The degree of hemolysis is an independent predictor for early mortality in SCD (Kato GJ et al, Blood 2006, Nouriae M et al, Haematologica 2013). There were progressively higher ratios of females:males with increasing age in SCD patients (Cochran's linear trend; UIC: P=0.03, Walk-PHaSST: P=0.005) (Figure 1).We compared gene expression in peripheral blood mononuclear cells by gender in 180 SCD patients from UIC using the Affymetrix Human Exon 2.0 ST array. Restricting analyses to autosomes with an FDR < 0.05, no statistically significant differences in gene expression were observed. We also determined the X-linked G6PD A- status by PCR in 259 SCD patients from the UIC cohort. G6PD A- deficiency was observed in 4% (6/155) of females and 10% (10/104) of males and was not associated with hemolytic markers.In summary, females had a lower degree of hemolysis, decreased vasculopathy, and represented a progressively higher proportion of SCD patients with increasing age in both the UIC and Walk-PHaSST cohorts. Future investigation of loci on the sex chromosomes for effects on Hb F levels and the degree of hemolysis are warranted. Understanding the mechanisms for reduced hemolysis in women with SCD, such as the potential effects of testosterone and estrogen on erythrocyte membrane stabilization, may also help guide future therapies. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Tocilizumab is safe and tolerable and reduces C-reactive protein concentrations in the plasma and cerebrospinal fluid of ALS patients.

We tested safety, tolerability, and target engagement of tocilizumab in amyotrophic lateral sclerosis (ALS) patients. Twenty-two participants, whose peripheral blood mononuclear cell (PBMC) gene expression profile reflected high messenger ribonucleic acid (mRNA) expression of inflammatory markers, were randomized 2:1 to three tocilizumab or placebo treatments (weeks 0, 4, and 8; 8mg/kg intravenous). Participants were followed every 4wk in a double-blind fashion for 16 wk and assessed for safety, tolerability, plasma inflammatory markers, and clinical measures. Cerebrospinal fluid (CSF) was collected at baseline and after the third treatment. Participants were genotyped for Asp358 Ala polymorphism of the interleukin 6 receptor (IL-6R) gene. Baseline characteristics, safety, and tolerability were similar between treatment groups. One serious adverse event was reported in the placebo group; no deaths occurred. Mean plasma C-reactive protein (CRP) level decreased by 88% in the tocilizumab group and increased by 4% in the placebo group (-3.0-fold relative change, P < .001). CSF CRP reduction (-1.8-fold relative change, P=.01) was associated with IL-6R C allele count. No differences in PBMC gene expression or clinical measures were observed between groups. Tocilizumab treatment was safe and well tolerated. PBMC gene expression profile was inadequate as a predictive or pharmacodynamic biomarker. Treatment reduced CRP levels in plasma and CSF, with CSF effects potentially dependent on IL-6R Asp358 Ala genotype. IL-6 trans-signaling may mediate a distinct central nervous system response in individuals inheriting the IL-6R C allele. These results warrant further study in ALS patients where IL-6R genotype and CRP levels may be useful enrichment biomarkers.

Effect of diet and nonesterified fatty acid levels on global transcriptomic profiles in circulating peripheral blood mononuclear cells in early lactation dairy cows

After calving, lipid mobilization caused by increased nutrient demands for lactation leads to elevated circulating concentrations of nonesterified fatty acids (NEFA). Excessive NEFA levels have previously been identified as a major risk factor for postpartum immunosuppression. The aim of this study was to investigate changes in global transcriptomic gene expression of peripheral blood mononuclear cells (PBMC) in dairy cows offered different early lactation diets (high concentrate, n = 7; medium, n = 8; or low, n = 9) and with differing circulating levels of NEFA. Cows were classified as having NEFA concentrations of either <500 µM (low, n = 6), 500 to 750 µM (medium, n = 8) or >750 µM (high, n = 10) at 14 d in milk. Plasma urea concentrations were greater for cows on the high concentrate diet but β-hydroxybutyrate and glucose concentrations did not differ significantly between either dietary treatments or NEFA groups. Cows with high NEFA weighed more at drying off and suffered greater body condition score loss after calving. The PBMC were isolated at 14 d in milk, and RNA was extracted for RNA sequencing. Differential gene expression was analyzed with DESeq2 with q-value for false discovery rate control followed by Gene Ontology Enrichment. Although there were no differentially expressed genes associated with lactation diet, 304 differentially expressed genes were identified between cows with high and low circulating NEFA, with 118 upregulated and 186 downregulated. Gene Ontology enrichment analysis demonstrated that biological adhesion and immune system process were foremost among various PBMC functions which were altered relating to body defenses and immunity. High NEFA concentrations were associated with inhibited cellular adhesion function by downregulating 20 out of 26 genes (by up to 17-fold) related to this process. Medium NEFA concentrations altered a similar set of functions as high NEFA, but with smaller enrichment scores. Localization and immune system process were most significant, with biological adhesion ranking only eleventh. Our results demonstrated that increased circulating NEFA concentrations, but not diet, were associated with immune system processes in PBMC in early lactation cows. Leukocyte cell-to-cell adhesion was inhibited when the NEFA concentration exceeded 750 µM, which would reduce the efficiency of diapedesis and so contribute to decreased body defense mechanisms and predispose animals to infection.

Gene expression changes and DNA damage after ex vivo exposure of peripheral blood cells to various CT photon spectra

Dual-energy CT provides enhanced diagnostic power with similar or even reduced radiation dose as compared to single-energy CT. Its principle is based on the distinct physical properties of low and high energetic photons, which, however, may also affect the biological effectiveness and hence the extent of CT-induced cellular damage. Therefore, a comparative analysis of biological effectiveness of dual- and single-energy CT scans with focus on early gene regulation and frequency of radiation-induced DNA double strand breaks (DSBs) was performed. Blood samples from three healthy individuals were irradiated ex vivo with single-energy (80 kV and 150 kV) and dual-energy tube voltages (80 kV/Sn150kV) employing a modern dual source CT scanner resulting in Volume Computed Tomography Dose Index (CTDIvol) of 15.79–18.26 mGy and dose length product (DLP) of 606.7–613.8 mGy*cm. Non-irradiated samples served as a control. Differential gene expression in peripheral blood mononuclear cells was analyzed 6 h after irradiation using whole transcriptome sequencing. DSB frequency was studied by 53BP1 + γH2AX co-immunostaining and microscopic evaluation of their focal accumulation at DSBs. Neither the analysis of gene expression nor DSB frequency provided any evidence for significantly increased biological effectiveness of dual-energy CT in comparison to samples irradiated with particular single-energy CT spectra. Relative to control, irradiated samples were characterized by a significantly higher rate of DSBs (p < 0.001) and the shared upregulation of five genes, AEN, BAX, DDB2, FDXR and EDA2R, which have already been suggested as radiation-induced biomarkers in previous studies. Despite steadily decreasing doses, CT diagnostics remain a genotoxic stressor with impact on gene regulation and DNA integrity. However, no evidence was found that varying X-ray spectra of CT impact the extent of cellular damage.

Effective DNA damage response after acute but not chronic immune challenge: SARS-CoV-2 vaccine versus Systemic Lupus Erythematosus

Whether and how an acute immune challenge may affect DNA Damage Response (DDR) is unknown. By studying vaccinations against Influenza and SARS-CoV-2 (mRNA-based) we found acute increases of type-I interferon-inducible gene expression, oxidative stress and DNA damage accumulation in blood mononuclear cells of 9 healthy controls, coupled with effective anti-SARS-CoV-2 neutralizing antibody production in all. Increased DNA damage after SARS-CoV-2 vaccine, partly due to increased oxidative stress, was transient, whereas the inherent DNA repair capacity was found intact. In contrast, in 26 patients with Systemic Lupus Erythematosus, who served as controls in the context of chronic immune activation, we validated increased DNA damage accumulation, increased type-I interferon-inducible gene expression and induction of oxidative stress, however aberrant DDR was associated with deficiencies in nucleotide excision repair pathways. These results indicate that acute immune challenge can indeed activate DDR pathways, whereas, contrary to chronic immune challenge, successful repair of DNA lesions occurs.

Gene Expression of PSORI-CM01 and Yinxieling in the Treatment of Psoriasis Vulgaris

Background. This study aimed to explore the mechanisms of action of the PSORI-CM01 and Yinxieling formulas in the treatment of patients with psoriasis vulgaris by analyzing gene expression in peripheral blood mononuclear cells (PBMCs). Methods. PBMC samples were collected from 21 patients before and after treatment. The study included nine patients in the PSORI-CM01 treatment group, 12 patients in the Yinxieling treatment group, and nine patients in the healthy control group. Gene expression levels in PBMCs were determined using the Affymetrix gene chip technology. Results. In the PSORI-CM01 group before and after treatment, a total of 668 differentially expressed genes were found, of which 445 were upregulated and 223 were downregulated. Before and after Yinxieling treatment, 657 differentially expressed genes were found, of which 168 were upregulated and 489 were downregulated. Venn analysis showed that 78 genes were not differentially expressed in the PSORI-CM01 group and 74 were not differentially expressed in the Yinxieling group compared with those in the controls. Among these genes, 72 genes were common to both groups, which were the genes on which the two drugs acted jointly. The results of KEGG analysis and Venn analysis on the signalling pathways of drug action in treatment groups showed that haemostasis and pathways involving Rho GTPases were common signalling pathways of drug action in the two groups. Conclusions. By a comparative analysis of the treatment groups, we found that both drugs have a positive effect on patients with psoriasis vulgaris, primarily by regulating the pathways related to platelet activation, aggregation, and blood coagulation. Trial registration: ChiCTR, ChiCTR-TRC-14005185, Registered 8 August 2014, http://www.chictr.org.cn/showproj.aspx?proj=4390