IL-10 Gene Expression Research Articles

The role of probiotics in restoring the Th1 to Th2 ratio in women experiencing recurrent implantation failure; a double-blind randomized clinical trial.

Among the complex causes of infertility, Recurrent Implantation Failure (RIF) stands out as a challenging condition. Immunological aberrations are closely implicated in RIF, particularly the imbalance between T helper 1 (Th1) and T helper 2 (Th2) cytokines. Interest in harnessing probiotics to enhance fertility outcomes in RIF-affected women is growing. This study aimed to investigate the potential of probiotics in restoring the Th1 to Th2 ratio and enhancing implantation outcomes in women experiencing RIF. A double-blinded randomized controlled trial enrolled 50 women with a record of at least three RIFs and 50 women with successful pregnancies as a control group in our study. RIF patients were randomly assigned to either the probiotic intervention group (n=25) or the placebo group (n=25). The probiotic group administered two tablets per day for 6months the probiotic tablets that contained prebiotic. At the same time, the placebo group received an identical-looking placebo. Flow cytometry technique was used to evaluate Th1 and Th2 cell frequencies. Gene expression of IL-4, IL-10, TNF-α and IFN-ɣ was analyzed using QRT-PCR. Serum concentrations of IL-4, IL-10, TNF-α and IFN-ɣ cytokines were evaluated using the ELISA technique. Flow cytometry analysis revealed significantly higher Th1 cell percentages in RIF patients compared to healthy controls, while healthy controls exhibited more significant Th2 cell proportions. Probiotic intervention led to a reduction in Th1 cells and an increase in Th2 cells in RIF patients. ELISA analysis indicated higher proinflammatory and lower anti-inflammatory cytokines compared to controls. After probiotic treatment, TNF-α and IFN-ɣ levels decreased, while IL-10 and IL-4 levels increased in RIF patients. QRT-PCR analysis showed no significant differences in cytokine gene expression between RIF patients before intervention. After probiotic therapy, pro-inflammatory cytokine expression decreased, and anti-inflammatory cytokine expression increased. Placebo treatment resulted in limited changes. Probiotic intervention effectively modulated the Th1 to Th2 ratio in RIF patients, evidenced by reduced Th1 cell percentages, enhanced Th2 cell populations, and cytokine level adjustments. The probiotic intervention improved implantation outcomes in women with RIF, as evidenced by a higher clinical pregnancy rate in the treatment group compared to the placebo group (p=0.037). These findings highlight the potential of probiotics in restoring immune balance and improving implantation outcomes in women with RIF.

FC24 Zasocitinib (TAK-279), an oral, allosteric, highly selective TYK2 inhibitor, modulates disease-central cytokine pathways that define clinical response in patients with moderate-to-severe plaque psoriasis

Abstract Zasocitinib (TAK-279) demonstrated promising efficacy and was generally well tolerated in a completed Phase 2b study (NCT04999839) in patients with moderate-to-severe plaque psoriasis, with the primary endpoint [75% improvement in psoriasis area and severity (PASI75)] achieved at doses ≥5 mg vs. placebo at Week 12. In an exploratory analysis of this study, associations between zasocitinib treatment, psoriasis/tyrosine kinase 2 (TYK2) biomarkers, and clinical/histological response were investigated. Lesional/non-lesional serum samples and skin biopsies were obtained with consent at Day 1 (pre-dose) and Weeks 4 and 12 from patients receiving zasocitinib (2, 5, 15, and 30 mg) or placebo orally once daily. RT-qPCR, RNA-seq and immunohistochemistry were used to assess changes in lesion keratinocyte proliferation (KRT16 expression), psoriasis/TYK2 biomarkers and lesion gene signatures, and their associations with clinical (PASI75) and histological (plaque resolution) response. Serum samples from 252 patients and biopsy samples from 63 patients were analysed. At Week 12, most clinical responders (n = 21/24, 88%) experienced reductions in KRT16 expression of >87% vs. baseline lesion levels. Pooled analysis showed that, in most histological responders (n = 18/25, 72%), interleukin (IL)17A and IL17F expression reduced by >80% vs. baseline lesion levels. Reductions in lesional type I interferon, IL-12 and IL-23 pathway gene expression were observed at zasocitinib 15 and 30 mg doses compared with baseline levels (P < 0.05). Dose- and time-dependent reductions in serum IL-17A, IL-17C and IL-17F were observed in all zasocitinib groups vs. placebo. In the 15 and 30 mg groups (n = 23), expression of key psoriasis/TYK2 biomarkers (e.g. DEFB4A, IL36G, IL23A) reverted to non-lesional levels at Week 12. Among evaluable PASI90 responders at Week 12 (n = 12), 48/50 of the top upregulated genes in lesions reverted to non-lesional expression levels. In all dose groups, patients had reduced lesional epidermal thickness, CD3+ T-cell counts and CD11c+ myeloid dendritic cell counts with zasocitinib vs. baseline. Zasocitinib treatment did not result in clinically meaningful longitudinal changes in laboratory parameters associated with Janus kinase inhibition. Zasocitinib modulated psoriasis/TYK2 biomarkers were associated with clinical and histological response in patients with moderate-to-severe plaque psoriasis.

Preface

As Editor-in-Chief of GenoMed Connect, it is my great pleasure to introduce the inaugural issue of this journal, dedicated to bridging the fields of genomics and medicine. Our vision for GenoMed Connect is to serve as a platform for the latest research that informs precision medicine and promotes integrative healthcare approaches. This inaugural issue features a diverse collection of seven articles, showcasing the journal's commitment to interdisciplinary research and its impact on advancing medical science. Each study addresses a unique aspect of genomics and medical research: 1. Evaluation of IL-12 Gene Expression in Coronary Artery Disease examines the role of IL-12 in atherosclerosis, shedding light on inflammatory pathways that may serve as therapeutic targets in coronary artery disease. 2. A Case Report on Epidermolytic Hyperkeratosis Type 1 introduces a novel mutation in the KRT1 gene, expanding the understanding of genetic dermatological conditions and offering insights into the molecular underpinnings of epidermolytic hyperkeratosis. 3. Identification of Prognostic Biomarkers in Uveal Melanoma explores tumor immune microenvironments, identifying genetic markers that could improve the prediction and treatment strategies for this rare but aggressive form of eye cancer. 4. HLA Class I and II Gene Diversity in North India provides an extensive genetic analysis that is valuable for transplantation immunology and population genetics, enhancing the knowledge of HLA variability and its implications in disease association. 5. Investigation of TLR and CD36 Expression in Coronary Artery Disease delves into the immune response mechanisms in coronary artery disease, examining the role of toll-like receptors and their potential as markers for disease progression. 6. Ethical Approaches in Reporting Secondary Findings from Exome Sequencing reviews current guidelines and ethical considerations, addressing challenges in reporting incidental findings and highlighting pathways for more responsible genomic data sharing. 7. The Role of Microbes in Endocrine Health presents an intriguing review on microbial endocrinology, exploring the influence of the gut microbiome on hormone regulation and its implications for metabolic health. These articles embody the interdisciplinary nature of GenoMed Connect, contributing to a deeper understanding of genomics in disease diagnosis, treatment, and ethical considerations. I extend my gratitude to our dedicated editorial board, the reviewers, and the team at Scifiniti for their hard work in bringing this issue to life. I look forward to witnessing the journal’s growth as it continues to foster collaboration and drive innovation in genomic medicine. I hope that the research presented in this issue inspires readers and advances our collective efforts to improve healthcare through genomic insights.

The effect of hyperbaric oxygen therapy on oxidative stress and inflammation in patients with diabetic foot ulcers: A Preliminary Study

IntroductionPatients with an uncontrolled glycemic index develop a wide variety of pathologies associated with diabetes, such as diabetic foot ulcers (DFUs). Hyperbaric oxygen therapy (HBOT) is an adjunctive therapy used to heal wounds and prevent lower extremity amputations in this population. ObjectiveThis preliminary study aimed to evaluate how HBOT impacts inflammation in patients with Wagner stages 2-4 DFUs by analyzing its effect on the gene expression of key oxidative stress regulators SOD1, SOD2, and GPX2, of pro-inflammatory cytokines TNFα, IL-1β, IL-4, and IL-12, and of the NLRP3 inflammasome. MethodsThe effect of HBOT was assessed in 15 patients with Wagner stages 2-4 DFUs that underwent 30 sessions in the hyperbaric chamber. This protocol is registered on Clinical Trials under the title Hyperbaric Oxygen Therapy in Diabetic Foot (15/07/2024) with the number NCT06502808. Blood samples were collected, and relative gene expression was assessed by quantitative real-time polymerase chain reaction (qPCR). ResultsThe hyperbaric chamber treatment increased the expression of SOD1 and GPX2 genes (0.4 and 3 times, respectively) after 30 sessions compared to baseline levels. Similarly, the gene expression of pro-inflammatory cytokines IL-1β, IL-12, IL-4, and NLRP3 increased after 30 sessions (2.1, 0.4, 1.5, and 1.2, respectively), while the expression of the TNFα gene decreased (0.5 times). Clinically, the patients’ lesions were fully resolved. ConclusionsHBOT directly influences the gene expression of several potent antioxidants and pro-inflammatory cytokines, thus favoring angiogenesis and blood circulation in the extremities.

Ceria-Stabilized Zirconia/Alumina Nanocomposite (NANO-Zr) Surface Enhances Osteogenesis Through Regulation of Macrophage Polarization

Zirconia implants are recognized for their excellent biocompatibility, aesthetics, and favorable mechanical properties. However, the effects of zirconia surfaces on osteogenesis, particularly in the presence of macrophages, are still not well understood. This study compares two types of zirconia surfaces—ceria-stabilized zirconia/alumina nanocomposite (NANO-Zr) and 3 mol% yttria-stabilized tetragonal zirconia polycrystal (3Y-TZP)—with titanium (Ti) substrates. Both zirconia surfaces promoted macrophage adhesion and proliferation, facilitated a shift from M1 to M2 polarization, and created an immune microenvironment conducive to osteogenesis by downregulating IL-6 and TNF-α and upregulating IL-10 and TGF-β gene expression. In macrophage co-cultures, both zirconia surfaces also supported osteoblast adhesion and proliferation, with NANO-Zr notably enhancing osteogenic differentiation and mineralization. These results highlight NANO-Zr as a promising candidate for future dental and orthopedic implant applications.

Influence of Feeding on IL-2 Gene Expression and Peak Blood Cyclosporine Concentration in Healthy Dogs Administered Oral Cyclosporine

Influence of Feeding on IL-2 Gene Expression and Peak Blood Cyclosporine Concentration in Healthy Dogs Administered Oral Cyclosporine

Expression of key cytokines in dog macrophages infected by Leishmania tarentolae opening new avenues for the protection against Leishmania infantum

The detection of Leishmania tarentolae in sympatric areas where Leishmania infantum is endemic raised questions regarding the protective effect exerted in dogs by L. tarentolae when in coinfection. This study aimed monitoring the in vitro gene expression of pro- (IFN- γ; TNF-α; IL-12) and anti-inflammatory (IL-4; IL-6; IL-10) cytokines in primary canine macrophages infected by L. tarentolae and L. infantum in single and in co-infections. Macrophages differentiated from dog blood mononuclear cells were infected with the L. tarentolae field-isolated (RI-325) and laboratory (LEM-124) strains, with L. infantum laboratory strain (IPT1), or both. Infection and the number of amastigotes per infected cell were evaluated microscopically by counting a total of 200 cells between 4 and 96 h. Cytokine gene expression was analyzed by real-time PCR from infected macrophages mRNA. Single infections presented higher expression of the cytokines IL-4 and IL-6, and lower of IL-12. Co-infections induced a lower gene expression of IL-4 and IL-6, and a higher gene expression of IL-12, correlating with the low amastigote burden despite the slight increase of infected cells. Data highlight the potential protective effect of L. tarentolae against L. infantum in co-infection by the reduced anti-inflammatory and increased pro-inflammatory cytokines gene expression, opening new perspectives for a canine vaccine development exploiting the non-pathogenic L. tarentolae.

Hypothetical proteins of chicken-isolated Limosilactobacillus reuteri subjected to in silico analyses induce IL-2 and IL-10

Lactic acid bacteria (LAB) probiotics are health-promoting but their characteristics, safety profile and functional mechanisms are not fully understood. Hence, this study aimed to characterize some hypothetical proteins of the chicken-isolated Limosilactobacillus reuteri genome and unravel their IL-2 and IL-10-inducing potential to understand mechanisms of their immunological functionality for sustainable applications. The selected proteins were subjected to in silico analyses for transmembrane topology, sub-cellular localization, IL-2 and IL-10-inducing ability and IL-2 and IL-10 gene expression across various conditions. IL-2 and IL-10-inducing mutants were statistically analyzed using a one-way analysis of variance of a general linear model of SAS and statistical significance was set at p < 0.05. The analyzed proteins are stable under a wide temperature range. All the hypothetical proteins are IL-2 and IL-10-inducing but QHPv.2.12, QHPv.2.13 and QHPv.2.15 are non-immunogenic. The evaluated mutants are IL-2 and IL-10-inducers but QHPv.2.13 and QHPv.2.15 are not immunogenic. This study sheds light on understanding the functional mechanisms of chicken-isolated L. reuteri and suggests it or its proteins as potent candidates for feed additive and therapeutic purposes.

Enrichment of Fat Graft with Association of ASC and Nanofat in an Animal Model.

Fat graft (FG) absorption rate varies from 20 to 80% in two years. Recently, several bioengineering techniques were applied to improve FG retention rate. Numerous studies investigated the use of adipocyte-derived stem cells (ASC) as FG enrichment. However, ASC production is costly, complex, and time-consuming. In contrast, Nanofat, a combination of lipids, stem cells and growth factors, offers a faster, simpler, and more cost-effective alternative for FG enrichment. This study aims to compare the effects of ASC with those of Nanofat, as a viable option in FG enrichment. Animals were allocated in three groups: Control group (1mL fat), ASC group (1mL fat +1x106ASC), andNnF group (1mL of fat + 0.3mL NnF). These groups were subdivided in three subgroups (4, 8, and 12 weeks, n = 6/group). We performed ultrasound and macroscopic measurements for FG volume, histology and expression of healing and inflammation genes. At week 12, ASC and NnF groups showed a higher retention of FG when compared to the Control group (51%, 46%, 12% respectively, p < 0.01). Fibrosis was similar in ASC and Nanofat groups. The Nanofat group showed a higher vascular density then the Control group (p < 0.05). Il-10 gene expression was higher, and Mmp9 was lower in the Nanofat group when compared to the ASC and Control groups. This study indicates that enriching FG with both ASC and Nanofat led to an increased retention rate of the FG, suggesting that Nanofat might be a promising alternative for FG enrichment. This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews, and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies, and Experimental Studies. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Clostridium sporogenes-derived metabolites protect mice against colonic inflammation

ABSTRACT Gut microbiota-derived metabolites play a pivotal role in the maintenance of intestinal immune homeostasis. Here, we demonstrate that the human commensal Clostridium sporogenes possesses a specific metabolic fingerprint, consisting predominantly of the tryptophan catabolite indole-3-propionic acid (IPA), the branched-chain acids (BCFAs) isobutyrate and isovalerate and the short-chain fatty acids (SCFAs) acetate and propionate. Mono-colonization of germ-free mice with C. sporogenes (CS mice) affected colonic mucosal immune cell phenotypes, including up-regulation of Il22 gene expression, and increased abundance of transcriptionally active colonic tuft cells and Foxp3+ regulatory T cells (Tregs). In DSS-induced colitis, conventional mice suffered severe inflammation accompanied by loss of colonic crypts. These symptoms were absent in CS mice. In conventional, but not CS mice, bulk RNAseq analysis of the colon revealed an increase in inflammatory and Th17-related gene signatures. C. sporogenes-derived IPA reduced IL-17A protein expression by suppressing mTOR activity and by altering ribosome-related pathways in Th17 cells. Additionally, BCFAs and SCFAs generated by C. sporogenes enhanced the activity of Tregs and increased the production of IL-22, which led to protection from colitis. Collectively, we identified C. sporogenes as a therapeutically relevant probiotic bacterium that might be employed in patients with inflammatory bowel disease (IBD).

Interleukin-1 Receptor-Associated Kinase 1 in Cancer Metastasis and Therapeutic Resistance: Mechanistic Insights and Translational Advances.

Interleukin-1 Receptor Associated Kinase 1 (IRAK1) is a serine/threonine kinase that plays a critical role as a signaling transducer of the activated Toll-like receptor (TLR)/Interleukin-1 receptor (IL-1R) signaling pathway in both immune cells and cancer cells. Upon hyperphosphorylation by IRAK4, IRAK1 forms a complex with TRAF6, which results in the eventual activation of the NF-κB and MAPK pathways. IRAK1 can translocate to the nucleus where it phosphorylates STAT3 transcription factor, leading to enhanced IL-10 gene expression. In immune cells, activated IRAK1 coordinates innate immunity against pathogens and mediates inflammatory responses. In cancer cells, IRAK1 is frequently activated, and the activation is linked to the progression and therapeutic resistance of various types of cancers. Consequently, IRAK1 is considered a promising cancer drug target and IRAK1 inhibitors have been developed and evaluated preclinically and clinically. This is a comprehensive review that summarizes the roles of IRAK1 in regulating metastasis-related signaling pathways of importance to cancer cell proliferation, cancer stem cells, and dissemination. This review also covers the significance of IRAK1 in mediating cancer resistance to therapy and the underlying molecular mechanisms, including the evasion of apoptosis and maintenance of an inflammatory tumor microenvironment. Finally, we provide timely updates on the development of IRAK1-targeted therapy for human cancers.

The Addition of Hot Water Extract of Juncao-Substrate Ganoderma lucidum Residue to Diets Enhances Growth Performance, Immune Function, and Intestinal Health in Broilers.

The purpose of this experiment was to investigate the effects of Hot Water Extract of Juncao-substrate Ganoderma lucidum Residue (HWE-JGLR) on the immune function and intestinal health of yellow-feather broilers. In an animal feeding experiment, 288 male yellow-feather broilers (1 day old) were randomly allocated to four treatment groups with six replicates of 12 birds each. The control (CON) group was fed a basal diet. HJ-1, HJ-2, and HJ-3 were fed a basal diet supplemented with 0.25%, 0.50%, and 1.00% HWE-JGLR, respectively. The feeding trial lasted for 63 d. The results showed increased ADFI (p = 0.033) and ADG (p = 0.045) of broilers in HJ-3, compared with the CON group. Moreover, higher contents of serum IL-4 and IL-10 and gene expression of IL-4 and IL-10 in jejunum mucosa and lower contents of serum IL-1β and gene expression of IL-1β in jejunum mucosa in HJ-3 were observed (p < 0.05). Additionally, the jejunal mucosal gene expression of Claudin-1 and ZO-1 in HJ-2 and HJ-3 was higher than that in the CON group (p < 0.05). As for the microbial community, compared with the CON group, the ACE index, Shannon index, and Shannoneven index of cecal microorganisms in HJ-2 and HJ-3 were elevated (p < 0.05). PCoA analysis showed that the cecal microbial structure of broilers in HJ-2 and HJ-3 was different from the CON group (p < 0.05). In contrast with the CON group, the broilers in HJ-2 and HJ-3 possessed more abundant Desulfobacterota at the phylum level and unclassified Lachnospiraceae, norank Clostridia vadinBB60 group and Blautia spp. at the genus level, while Turicibacter spp. and Romboutsia spp. were less (p < 0.05). In conclusion, dietary supplementation with HWE-JGLR can improve growth performance, enhance body immunity and intestinal development, and maintain the cecum microflora balance of yellow-feather broilers.

Investigation of the relationship of tissue-resident γδ T cells and IL-17 gene expression with the pathogenesis of autoimmune hepatitis.

Autoimmune hepatitis (AIH) is a chronic inflammatory liver disease. Elevated serum immunoglobulin G (IgG) levels, autoantibodies, and histopathological interface hepatitis are the hallmarks of AIH. Autoantibodies and pathological findings, clinical and biochemical features, typical immunoglobulin levels, and exclusion of other diseases are used to diagnose the condition. Gamma-delta (γδ) T cells are a unique population of unconventional T cells with γ and δ glycoprotein chains. γδ T cells have been shown to play a crucial role in autoimmune diseases by producing interleukin (IL)-17. However, its role in AIH remains to be further elucidated. In this study, we aimed to examine the role of γδ T cells and IL-17 in the pathogenesis of AIH, by working on biopsy samples. Paraffin blocks of 18 patients with type 1 AIH and 18 control liver tissues were analyzed. qRT-PCR assessed IL-17 gene expression. Immunofluorescence double staining of CD3+TCRγδ+ was performed to reveal tissue-resident γδ T cells' role in AIH. When comparing AIH to the control, there was a substantial increase in the ratio of CD3+TCRγδ+ cells in total inflammatory cells (p = 0.01). IL-17 gene expression was lowered in AIH when compared to the control (p = 0.01). This study provides evidence for the involvement of γδ T cells and IL-17 in the pathogenesis of AIH. The ratio of γδ T cells and IL-17 gene expression showed a significant difference in AIH suggesting a potential role for γδ T cells in driving liver inflammation in A fIH.

Scutellaria baicalensis ameliorates allergic airway inflammation through agonism and transcriptional regulation of TAS2Rs

Ethnopharmacological relevanceScutellaria baicalensis Georgi (SCB, Huangqin) is a traditional medicinal plant used to treat fever and respiratory diseases. SCB has a good therapeutic effect on asthma and anti-inflammation in traditional clinic use. However, the molecular mechanism and targets of SCB in treating asthma are still unclear. Aim of the studyCombining transcriptomic analysis and in vitro experimental validation, this study aimed to reveal the molecular mechanism and targets of SCB in treating asthma. Materials and methodsThe anti-asthmatic effects of SCB and its active components, scutellarin and oroxylin A, were evaluated in ovalbumin (OVA)-induced rats by analysis of pulmonary function and pathology. The signaling pathways in rat pulmonary tissue were analyzed using transcriptomics and protein interaction network analysis. Calcium mobilization assay and molecular docking were utilized to discover the active compounds from SCB with agonism activity of type 2 taste receptors (TAS2Rs). The anti-asthmatic effect and transcriptional regulation of TAS2Rs regulated by SCB and its active components were analyzed in vitro. ResultsExtracts of SCB (ESB), scutellarin, and oroxylin A ameliorated airway function and inflammation in OVA-induced rats. The anti-asthma mechanism of ESB, scutellarin and oroxylin A was highly related to immune and taste transduction pathways based on transcriptomic analysis, especially the TAS2Rs signaling pathway. ESB was the direct agonist of TAS2R4 and TAS2R14 with EC50 of 209.1 and 217.2 μg/mL based on calcium mobilization assay, respectively. Baicalein was the main active component for TAS2R4 agonism activity, and scutellarin and oroxylin A had weak agonism activity of TAS2R4 and TAS2R14 through calcium mobilization assay and molecular docking. However, scutellarin and oroxylin A significantly upregulated the gene expression of Tas2r108 (the mouse ortholog of the TAS2R4) in lung tissue. ESB, scutellarin, and oroxylin A inhibited LPS-induced lactate dehydrogenase release and gene expression of TNF through transcriptional regulation of TAS2R4 and TAS2R14 on bronchial epithelial cells. ESB and oroxylin A ameliorated IgE-induced β-hexosaminidase release and gene expression of Il4 and Tnf and upregulated gene expression of Tas2r108. ConclusionThese results provided new insight into the anti-asthmatic mechanism of SCB and active components, scutellarin and oroxylin A, through agonism and transcriptional regulation of TAS2Rs to ameliorate allergic airway inflammation.

The lipooligosaccharide of the gut symbiont Akkermansia muciniphila exhibits a remarkable structure and TLR signaling capacity

The cell-envelope of Gram-negative bacteria contains endotoxic lipopolysaccharides (LPS) that are recognized by the innate immune system via Toll-Like Receptors (TLRs). The intestinal mucosal symbiont Akkermansia muciniphila is known to confer beneficial effects on the host and has a Gram-negative architecture. Here we show that A. muciniphila LPS lacks the O-polysaccharide repeating unit, with the resulting lipooligosaccharide (LOS) having unprecedented structural and signaling properties. The LOS consists of a complex glycan chain bearing two distinct undeca- and hexadecasaccharide units each containing three 2-keto-3-deoxy-D-manno-octulosonic acid (Kdo) residues. The lipid A moiety appears as a mixture of differently phosphorylated and acylated species and carries either linear or branched acyl moieties. Peritoneal injection of the LOS in mice increased higher gene expression of liver TLR2 than TLR4 (100-fold) and induced high IL-10 gene expression. A. muciniphila LOS was found to signal both through TLR4 and TLR2, whereas lipid A only induced TLR2 in a human cell line. We propose that the unique structure of the A. muciniphila LOS allows interaction with TLR2, thus generating an anti-inflammatory response as to compensate for the canonical inflammatory signaling associated with LOS and TLR4, rationalizing its beneficial host interaction.

Dynamic chromatin architecture identifies new autoimmune-associated enhancers for IL2 and novel genes regulating CD4+ T cell activation.

Genome-wide association studies (GWAS) have identified hundreds of genetic signals associated with autoimmune disease. The majority of these signals are located in non-coding regions and likely impact cis-regulatory elements (cRE). Because cRE function is dynamic across cell types and states, profiling the epigenetic status of cRE across physiological processes is necessary to characterize the molecular mechanisms by which autoimmune variants contribute to disease risk. We localized risk variants from 15 autoimmune GWAS to cRE active during TCR-CD28 co-stimulation of naïve human CD4+ T cells. To characterize how dynamic changes in gene expression correlate with cRE activity, we measured transcript levels, chromatin accessibility, and promoter-cRE contacts across three phases of naive CD4+ T cell activation using RNA-seq, ATAC-seq, and HiC. We identified ~1200 protein-coding genes physically connected to accessible disease-associated variants at 423 GWAS signals, at least one-third of which are dynamically regulated by activation. From these maps, we functionally validated a novel stretch of evolutionarily conserved intergenic enhancers whose activity is required for activation-induced IL2 gene expression in human and mouse, and is influenced by autoimmune-associated genetic variation. The set of genes implicated by this approach are enriched for genes controlling CD4+ T cell function and genes involved in human inborn errors of immunity, and we pharmacologically validated eight implicated genes as novel regulators of T cell activation. These studies directly show how autoimmune variants and the genes they regulate influence processes involved in CD4+ T cell proliferation and activation.

Blocking IL-23 Signaling Mitigates Cigarette Smoke-Induced Murine Emphysema.

Inflammatory cell infiltration is a characteristic feature of COPD and correlates directly with the severity of the disease. Interleukin-23 (IL-23) is a pro-inflammatory cytokine that regulates Th-17 inflammation, which mediates many pathophysiological events in COPD. The primary goal of this study was to determine the role of IL-23 as a mediator of key pathologic processes in cigarette smoke-induced COPD. In this study, we report an increase in IL23 gene expression in the lung biopsies of COPD patients compared to controls and identified a positive correlation between IL23 gene expression and disease severity. In a cigarette smoke-induced murine emphysema model, the suppression of IL-23 with a monoclonal blocking antibody reduced the severity of cigarette smoke-induced murine emphysema. Mechanistically, the suppression of IL-23 was associated with a reduction in immune cell infiltration, oxidative stress injury, and apoptosis, suggesting a role for IL-23 as an essential immune mediator of the inflammatory processes in the pathogenesis of CS-induced emphysema.

Alterations in CD4+ T Cell Cytokines Profile in Female Patients with Hashimoto's Thyroiditis Following Vitamin D Supplementation: A Double-blind, Randomized Clinical Trial.

Hashimoto's thyroiditis (HT) is an autoimmune disease characterized by the destruction of thyroid cells through immune processes involving T helper (Th)1 cytokines. This clinical trial investigates the impact of vitamin D supplementation on serum cytokine levels and gene expression in CD4+ T cells from HT patients, aiming to understand its effects on Th-1, Th-2, Th-17, and regulatory T (Treg) cell-associated factors. Female patients were randomly assigned in a double-blind design to either a vitamin D-supplemented group, which received cholecalciferol (1, 25(OH)2D3) at a dose of 50,000 IU, or the placebo group, which received a weekly placebo for a duration of three months. Serum cytokine levels were assessed using enzyme-linked immunosorbent assay (ELISA), while genes' expression levels were measured using real-time PCR. Serum 25-hydroxyvitamin D and levels exhibited a significant increase following vitamin D supplementation, in comparison to the placebo group. Additionally, the vitamin D supplementation resulted in a significant elevation of serum calcium (Ca) levels compared to baseline. In the vitamin D group, there was a significant decrease in both serum levels and expression of the interleukin (IL)-17 gene when compared to baseline, although no statistical difference was observed between the placebo and vitamin D groups. The gene expression of transforming growth factor-beta (TGFβ) was significantly increased in the vitamin D group compared to baseline, with no significant difference between the two study groups. Vitamin D treatment had no effect on serum levels of interferon-gamma (IFNϒ) and IL-4. While the gene expression of IL-4 in the vitamin D group did not exhibit a statistically significant increase, the level of GATA3 transcription factor increased significantly when compared to the placebo group. The expression of IFNϒ and transcription factors, T-bet, RORc, and forkhead box protein 3 (FOXP3) in genes did not show significant changes following vitamin D supplementation. The findings suggest that vitamin D supplementation may hold potential benefits for autoimmune diseases, such as HT. However, further longitudinal clinical trials are necessary to gain a more comprehensive understanding of the specific effects of vitamin D on HT.

Shedding reduction and immunity modulation in piglets with an inactivated Mycoplasma hyopneumoniae vaccine encapsulated in nanostructured SBA-15 silica

Mycoplasma (M.) hyopneumoniae is a primary etiological agent of porcine enzootic pneumonia (PEP), a disease that causes significant economic losses to pig farming worldwide. Current commercial M. hyopneumoniae vaccines induce partial protection, decline in preventing transmission of this pathogen or inducing complete immunity, evidencing the need for improving vaccines against PEP. In our study, we aimed to test the effectiveness of the SBA-15 ordered mesoporous silica nanostructured particles as an immune adjuvant of a vaccine composed of M. hyopneumoniae strain 232 proteins encapsulated in SBA-15 and administered by intramuscular route in piglets to evaluate the immune responses and immune-protection against challenge. Forty-eight 24-day-old M. hyopneumoniae-free piglets were divided into four experimental groups with different protocols, encompassing a commercial vaccine against M. hyopneumoniae, SBA-15 vaccine, SBA-15 adjuvant without antigens and a non-immunized group. All piglets were challenged with the virulent strain 232 of M. hyopneumoniae. Piglets that received the SBA-15 and commercial vaccine presented marked immune responses characterized by anti-M. hyopneumoniae IgA and IgG antibodies in serum, anti-M. hyopneumoniae IgA antibodies in nasal mucosa and showed an upregulation of IL-17 and IL-4 cytokines and downregulation of IFN-γ in lungs 35 days post-infection. Piglets immunized with SBA-15 vaccine presented a reduction of bacterial shedding compared to piglets immunized with a commercial bacterin. In addition, piglets from SBA-15 adjuvant suspension group presented increased IL-17 gene expression in the lungs without involvement of Th1 and Th2 responses after challenge. These results indicated that SBA-15 vaccine induced both humoral and cell-mediated responses in the upper respiratory tract and lungs, first site of replication and provided protection against M. hyopneumoniae infection with a homologous strain with reduction of lung lesions and bacterial shedding. Finally, these results enhance the potential use of new technologies such as nanostructured particles applied in vaccines for the pig farming industry.

Conditioned media from human adipose tissue-derived mesenchymal stem cells: potential effect on peripheral blood mononuclear cells in co-culture with HeLa cell line.

The use of mesenchymal stem cells (MSCs) for the treatment of various diseases is being investigated, however, their use in cervical cancer has not been well-studied. Here, we examined the impact of collected MSC-conditioned medium (CM) on 1 to 5 days on apoptosis, proliferation, and cytokine production of peripheral blood mononuclear cells (PBMCs) when co-cultured alongside the HeLa cell line for 24, 48, and 72h by CFSE assay, flow cytometry, and real-time PCR, respectively. We found that CMs collected on the third day of MSCs culture significantly increased the proliferation of PBMCs and decreased the proliferation of HeLa cells after 48h. CMs showed no significant effects on cell death, whereas it significantly increased the apoptosis of HeLa cells. Real-time PCR analysis showed that the presence of CM collected on the third day of MSCs culture caused a significant increase in the gene expression of IL2, IFN-γ, and TGF-β in PBMCs after 48h co-culture with HeLa cells. The data mentioned earlier demonstrate that MSC-CM can induce the growth and endurance of PBMCs while concurrently culturing HeLa cells. This observation indicates their promising potential as immunomodulatory therapies for cervical cancer cells. Nevertheless, additional investigation is imperative to comprehensively comprehend the fundamental mechanisms and refine therapeutic strategies involving PBMCs and mesenchymal stem cells.