IL-10 Gene Expression Research Articles

Cannabinoid receptor type 2 is upregulated in synovium following joint injury and mediates anti-inflammatory effects in synovial fibroblasts and macrophages

Joint injury-induced perturbations to the endocannabinoid system (ECS), a regulator of both inflammation and nociception, remain largely uncharacterized. We employed a mouse model of ACL rupture to assess alterations to nociception, inflammation, and the ECS while using in vitro models to determine whether CB2 agonism can mitigate inflammatory signaling in macrophages and fibroblast-like synoviocytes (FLS). Mice underwent noninvasive ACL rupture (ACLR) via tibial compression-based loading. Nociception was measured longitudinally using mechanical allodynia and knee hyperalgesia testing. Synovitis was assessed using histological scoring and histomorphometry. Gene and protein markers of inflammation were characterized in whole joints and synovium. Immunohistochemistry assessed injury-induced alterations to CB1+, CB2+, and F4/80+ cells in synovium. To assess whether CB2 agonism can inhibit pro-inflammatory macrophage polarization, murine bone marrow-derived macrophages (mBMDM) were stimulated with IL-1β or conditioned medium from IL-1β-treated FLS and treated with vehicle (DMSO), the CB2 agonist HU308, or cannabidiol (CBD). Macrophage polarization was assessed as the ratio of M1-associated (IL1b, MMP1b, and IL6) to M2-associated (IL10, IL4, and CD206) gene expression. Human FLS (hFLS) isolated from synovial tissue of OA patients were treated with vehicle (DMSO) or HU308 following TNF-α or IL-1β stimulation to assess inhibition of catabolic/inflammatory gene expression. ACLR induces synovitis, progressively-worsening PTOA severity, and an immediate and sustained increase in both mechanical allodynia and knee hyperalgesia, which persist beyond the resolution of molecular inflammation. Enrichment of CB2, but not CB1, was observed in ACLR synovium at 3d, 14d, and 28d, and CB2 was found to be associated with F4/80 (+) cells, which are increased in number in ACLR synovium at all time points. The CB2 agonist HU308 strongly inhibited mBMDM M1-type polarization following stimulation with either IL-1β or conditioned medium from IL-1β-treated mFLS, which was characterized by reductions in Il1b, Mmp1b, and Il6 and increases in Cd206 gene expression. Cannabidiol similarly inhibited IL-1β-induced mBMDM M1 polarization via a reduction in Il1b and an increase in Cd206 and Il4 gene expression. Lastly, in OA hFLS, HU308 treatment inhibited IL-1β-induced CCL2, MMP1, MMP3, and IL6 expression and further inhibited TNF-α-induced CCL2, MMP1, and GMCSF expression, demonstrating human OA-relevant anti-inflammatory effects by targeting CB2. Joint injury perturbs the intra-articular ECS, characterized by an increase in synovial F4/80(+) cells, which express CB2, but not CB1. Targeting CB2 in murine macrophages and human FLS induced potent anti-inflammatory and anti-catabolic effects, which indicates that the CB2 receptor plays a key role in regulating inflammatory signaling in the two primary effector cells in the synovium. The intraarticular ECS is therefore a potential therapeutic target for blocking pathological inflammation in future disease-modifying PTOA treatments.

Effect of nucleotides on growth performance, gut health, and some immunological parameters of broiler chicken exposed to high stocking density

The current study was conducted to investigate the effect of dietary nucleotides inclusion rate on growth performance, gut health, and immunity of broiler chickens exposed to high stocking density-induced stress. A total of 315 one-day-old mixed broiler chickens were randomly assigned to a 2 × 3 factorial arrangement of treatments with 2 different stocking densities, low stocking density (LSD; 0.073 m2/2.2 kg broiler chicken; 15 broiler chickens/replicate) and high stocking density (HSD; 0.05 m2/2.2 kg broiler chicken; 20 broiler chickens/replicate), and 3 dietary inclusion rates of nucleotides at 0, 0.05, and 0.1%, with 3 cages per treatment. The experiment lasted for 35 days. At the end of the experiment, intestinal histomorphology, digestive enzymes production, immune status, and growth performance of broiler chickens were determined. The results showed that intestinal histomorphological picture, immune indices, and digestive enzymes production were negatively affected (P < 0.05) with HSD, resulting in a decreased (P < 0.001) body weight gain (BWG), feed intake (FI) (P < 0.001), and an increased (P < 0.001) feed conversion ratio (FCR). While the intestinal and pancreatic enzymes production and intestinal histomorphology, including villus height, mucosal thickness, and goblet cell number increased linearly (P < 0.001) with the increased nucleotides supplementation rate, which resulted in a linear increase in BWG (P < 0.001) and FI (P < 0.001), and a decrease in FCR (P = 0.026). However, the effect was more prominent at LSD than HSD. In addition, immune indices, including antibody against NDV, IL-2 and INF-γ gene expression, and lysozyme production, linearly increased (P < 0.001) with the increase of dietary nucleotides inclusion rate, resulting in partial mitigation of immune compromise in broiler chickens at HSD. We concluded that dietary nucleotide supplementation, particularly at 0.1%, partially attenuated the negative effect of HSD on broiler chicken performance through improving intestinal histomorphology, digestive enzyme production, and immune status.

Network pharmacology-based study on immunomodulatory mechanism of danggui-yimucao herb pair for the treatment of RU486-induced abortion

Ethnopharmacological relevanceThe Danggui-Yimucao herb pair (DY) is a classic combination in Chinese herbal formulas, consisting of the root of Angelica sinensis (Oliv.) Diels and the aerial parts of Leonurus japonicus Houtt. DY first appeared in “Zhulinsi fuke mifang” in the Jin Dynasty, and it has a long history as a drug for the treatment of abortion. However, its underlying immunomodulatory mechanisms involved are still unclear. Aim of the studyIn this study, network pharmacology and pharmacological experiments were used to explore the role and mechanism of DY in the treatment of medical abortion. Materials and methodsNetwork pharmacology was used to establish the relationship between the components of DY and abortion-related targets, and to enrich important pathways and biological process for verification. ELISA was used to assess progesterone levels. Flow cytometry was used to detect the degree of differentiation of Th1/Th2 cells. Immunohistochemical methods and qPCR were used to measure the expression levels of T-bet, GATA-3 and IL-4. ResultsThrough the prediction analysis of network pharmacology, we found that key pathway for DY treatment of abortion, such as anemia, pelvic infection, immune disorders, and coagulation disorders, was Th1/Th2 cell differentiation pathway. The pharmacological results revealed that DY greatly corrected the imbalance of Th cell subsets in abortion mice, significantly inhibited the differentiation of Th2 cells, and resulted in an increase in the Th1/Th2 ratio. In addition, the concentration of progesterone in the serum of mice after abortion was significantly reduced. We also found that DY upregulated spleen T-bet and downregulated IL-4 gene expression in mice. Besides, immunohistochemical results showed that DYE could up-regulate T-bet but inhibit GATA-3 expression. ConclusionsOur results showed that after RU486-induced abortion, progesterone and Th1/Th2 paradigm were disordered in mice, but DY could make mice recover more quickly, which indicated that DY had great development value in immunoregulation.

Sex differences and similarities in the neuroimmune response to central administration of poly I:C

BackgroundThe neuroimmune system is required for normal neural processes, including modulation of cognition, emotion, and adaptive behaviors. Aberrant neuroimmune activation is associated with dysregulation of memory and emotion, though the precise mechanisms at play are complex and highly context dependent. Sex differences in neuroimmune activation and function further complicate our understanding of its roles in cognitive and affective regulation.MethodsHere, we characterized the physiological sickness and inflammatory response of the hippocampus following intracerebroventricular (ICV) administration of a synthetic viral mimic, polyinosinic:polycytidylic acid (poly I:C), in both male and female C57Bl/6N mice.ResultsWe observed that poly I:C induced weight loss, fever, and elevations of cytokine and chemokines in the hippocampus of both sexes. Specifically, we found transient increases in gene expression and protein levels of IL-1α, IL-1β, IL-4, IL-6, TNFα, CCL2, and CXCL10, where males showed a greater magnitude of response compared with females. Only males showed increased IFNα and IFNγ in response to poly I:C, whereas both males and females exhibited elevations of IFNβ, demonstrating a specific sex difference in the anti-viral response in the hippocampus.ConclusionOur data suggest that type I interferons are one potential node mediating sex-specific cytokine responses and neuroimmune effects on cognition. Together, these findings highlight the importance of using both males and females and analyzing a broad set of inflammatory markers in order to identify the precise, sex-specific roles for neuroimmune dysregulation in neurological diseases and disorders.

Influence of Intracanal Cryotherapy on Inflammatory Mediators Expression using Different Irrigation Protocols: A Randomized Clinical Trial

ABSTRACTAim: To evaluate the effect of intra-canal cryotherapy on interleukin-1βand interleukin-10 expression in teeth with symptomatic apical periodontitis.Methodology: Forty-eight single rooted teeth in patients with symptomatic apical periodontitiswere randomly divided into 4 groups according to the irrigation protocol.Group I (control); 5% sodium hypochlorite at room temperature. Group II; 5% coldsodium hypochlorite (2-5°C). Group III; final rinse with 20 mL of 5% cold sodiumhypochlorite (2-5°C) for 5 minutes. Group IV; final rinse with 20 mL of cold saline (2-5°C) for 5 minutes. Paper point passing 2 mm beyond the apex was used to collect periapicalfluid before, immediately after mechanical instrumentation and after one week tocharacterize the mRNA expression of IL-1β and Il-10 using real-time polymerase chainreaction (PCR). Statistical analysis was performed using one-way ANOVA to comparebetween all groups. Results: Our results revealed statistically significant (at P≤ 0.05)down regulation of IL-1β gene expression levels in all groups after one week with nostatistically significant difference between the four groups and statistically significant(at P≤ 0.05) up regulation of IL-10 gene expression levels in all groups after one week.Conclusion: All the cryotherapy irrigation protocols showed lower levels of expressionof the pro-inflammatory mediators (interleukin-1β) and higher levels of expression ofthe anti-inflammatory mediator (interleukin-10) than the control group after one week.

Autophagy-Associated IL-15 Production Is Involved in the Pathogenesis of Leprosy Type 1 Reaction.

Leprosy reactional episodes are acute inflammatory events that may occur during the clinical course of the disease. Type 1 reaction (T1R) is associated with an increase in neural damage, and the understanding of the molecular pathways related to T1R onset is pivotal for the development of strategies that may effectively control the reaction. Interferon-gamma (IFN-γ) is a key cytokine associated with T1R onset and is also associated with autophagy induction. Here, we evaluated the modulation of the autophagy pathway in Mycobacterium leprae-stimulated cells in the presence or absence of IFN-γ. We observed that IFN-γ treatment promoted autophagy activation and increased the expression of genes related to the formation of phagosomes, autophagy regulation and function, or lysosomal pathways in M. leprae-stimulated cells. IFN-γ increased interleukin (IL)-15 secretion in M. leprae-stimulated THP-1 cells in a process associated with autophagy activation. We also observed higher IL15 gene expression in multibacillary (MB) patients who later developed T1R during clinical follow-up when compared to MB patients who did not develop the episode. By overlapping gene expression patterns, we observed 13 common elements shared between T1R skin lesion cells and THP-1 cells stimulated with both M. leprae and IFN-γ. Among these genes, the autophagy regulator Translocated Promoter Region, Nuclear Basket Protein (TPR) was significantly increased in T1R cells when compared with non-reactional MB cells. Overall, our results indicate that IFN-γ may induce a TPR-mediated autophagy transcriptional program in M. leprae-stimulated cells similar to that observed in skin cells during T1R by a pathway that involves IL-15 production, suggesting the involvement of this cytokine in the pathogenesis of T1R.

Targeting CD166+ lung cancer stem cells: Molecular study using murine dendritic cell vaccine

PurposeCancer stem cells (CSC) are the most common causes of lung cancer relapse and mouse resistance to chemotherapy. CD166 was identified as CSC marker for lung cancer. Our study aimed to detect the effect of dendritic cell vaccine loaded with tumor cell lysate (TCL-DCV) on percentage of CD166+ CSC in lung of mice exposed to Benzo(a)Pyrene (BP). MethodsFemale albino mice were divided into 5 groups (22 mice per group): normal control (NC), lung cancer control (LCC) (50 mg/kg BP orally, twice weekly for four weeks), dendritic cell (DC), TCL-DCV and cisplatin. Cisplatin (6 mg/kg, intraperitoneal) was given in two doses (18th and 20th week). 1 × 106 cells of each of DC and TCL-DCV was given subcutaneously as cisplatin. At the end of experiment (22 weeks), lung tissue was used for evaluation of cytotoxic T lymphocyte antigen-4 (Ctla-4), transforming growth factor-β (Tgf-β), forkhead box protein P3 (Foxp3), programmed death ligand 1 (Pd-l1) and interleukin 12 (Il-12) gene expression using quantitative RT-PCR. The percentage of CD83+, CD8+ and CD166+ cells in lung tissue were measured using flow cytometry. ResultsThe results revealed that TCL-DCV reversed the tumorigenic effect of BP in the lung as evidenced by histopathological examination. Compared to cisplatin, dendritic cell vaccination (TCL-DCV) significantly decreased percentage of CD166+ CSC. This anticancer stemness effect was attributed to the immune-stimulatory effect as indicated by increased percentage of CD83+ and CD8+ cells, upregulation of Il-12, and downregulation of Tgf-β, Ctla-4, Pd-l1 and Foxp3 gene expression compared to LCC group. ConclusionsTCL-DCV ameliorated cancer stemness through modulating tumor immune archetypes which make it a potent therapeutic alternative to chemotherapy resistant cases.

Short-term transport stress and supplementation alter immune function in aged horses.

Long-distance transport is associated with stress-related changes in equine immune function, and shipping-associated illnesses are often reported. Horses are frequently transported short distances, yet the effects of short-term transport on immune function remain largely unknown. Twelve horses, aged 15–30 yr, were assigned to either the control (n = 6) or treatment (n = 6) groups; treatment horses received a daily antioxidant supplement 3 weeks before and after transport. All horses were transported for approximately 1.5–2 hr on Day 0. Blood was collected via jugular venipuncture at 15-min pre- and post-transport and on Days –21, 1, 3, 7, 14, and 21. Body temperature, heart rate, body weight, total cortisol, and gene expression of IFNγ, IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12α, IL-17α, SAA1, and TNFα in whole blood were measured. Peripheral blood mononuclear cells were isolated, stimulated with PMA/ionomycin, and stained for IFNγ and TNFα before analysis via flow cytometry. Statistical analyses were performed with significance set at P < 0.05 (SAS 9.4). Transport and supplementation did not appear to affect body weight, heart rate, IL-4, IL-8, IL-12α, IL-17α, change (Δ) in the % and mean fluorescence intensity (MFI) of IFNγ+ lymphocytes after stimulation, or Δ in the % and MFI of TNFα+ lymphocytes after stimulation. Supplementation decreased IL-1β and SAA1 expression. Transport increased total cortisol concentration, body temperature, and IL-2, IL-6, and IL-10 expression but decreased IL-1β, TNFα, and IFNγ expression. Short-term transportation affected physiological, endocrine, and immune responses; supplementation may ameliorate inflammation in aged horses. Immune responses were most altered at 15-min post-transport and typically recovered by Day 1, suggesting that horses may be vulnerable to disease during and almost immediately after short-term transport.

The impact of cryopreservation in signature markers and immunomodulatory profile of tendon and ligament derived cells.

Tendon and ligament (T/L) engineering strategies towards clinical practice have been challenged by a paucity of understanding in the identification and still poorly described characterization of cellular niches. Prospecting how resident cell populations behave in vitro, and how cryopreservation may influence T/ L-promoting factors, can provide insights intoT/ L-cellular profiles for novel regenerative solutions. Therefore, we studiedhuman T/ L-derived cells isolated from patellar tendons and cruciate ligaments as suitable cellular models to anticipate tendon and ligament niches responses for advanced strategies with predictive tenogenic and ligamentogenic value. Our results show that the crude populations isolated from tendon and ligament tissues hold a stem cell subset and share a similar behavior in terms of tenogenic/ligamentogenic commitment. Both T/ L-derived cells successfully undergo cryopreservation/thawing maintaining the tenogenic/ligamentogenic profiles. The major differences between cryopreserved and fresh populations were observed at the gene expression of MKX, SCX, and TNMD as well as at the protein levels of collagen type I and III, in which cells from tendon origin (hTDCs) evidence increased values in comparison to the ones from ligament (hLDCs, p < 0.05). In addition, low-temperature storage was shown to potentiate an immunomodulatory profile of cells, especially in hTDCs leading to an increase in the gene expression of the anti-inflammatory factors IL-4 and IL-10 (p < 0.05), as well as in the protein secretion of IL-10 (p < 0.01) and IL-4 (p < 0.001). Overall, the outcomes highlight the relevance of the cryopreserved T/ L-derived cells and their promising immunomodulatory cues as in vitro models for investigating cell-mediated mechanisms driving tissue healing and regeneration.

Anti-Inflammatory Activity of S. Marianum and N. Sativa Extracts on Macrophages.

Nigella sativa (N. sativa) and Silybum marianum (S. marianum) are used to regulate macrophage polarization in lipopolysaccharide-induced RAW 264.7 cells and thioglycollate-elicited peritoneal inflammation. Cytotoxicity assays and acute toxicity tests were performed to investigate the safe dose and toxicity of the prepared extracts. Also, nitric oxide production was determined by Griess assay on RAW264.7 and peritoneal macrophage supernatants. After RNA extraction from macrophages, real-time PCR was performed to measure the relative gene expression of tumor necrosis factor (TNF)-α, interleukin (IL)-6, transforming growth factor (TGF)-β, and IL-10. Finally, regulatory T cells (Treg cells) were counted by flow cytometry. S. marianum methanolic extract (SME), N. sativa ethanolic extract (NEE), and their mixture (SME+NEE) decreased NO levels significantly in RAW264.7 and peritoneal murine macrophages. N. sativa ethanolic extract significantly increased IL-10 gene expression and significantly decreased IL-6 and TNF-α expression in RAW264.7 cells. In mixture-treated peritoneal macrophages, IL-10 and TGF-β expression were significantly increased, while IL-6 and TNF-α were significantly decreased. Also, the percentage of Treg cells was significantly greater in the mixture-treated cells than in controls. These results suggest that an SME and NEE mixture has anti-inflammatory and immunomodulatory activities and may be useful in the treatment of diseases of immunopathologic origin characterized by macrophage hyperactivation.

Microbiota Signals during the Neonatal Period Forge Life-Long Immune Responses.

The microbiota regulates immunological development during early human life, with long-term effects on health and disease. Microbial products include short-chain fatty acids (SCFAs), formyl peptides (FPs), polysaccharide A (PSA), polyamines (PAs), sphingolipids (SLPs) and aryl hydrocarbon receptor (AhR) ligands. Anti-inflammatory SCFAs are produced by Actinobacteria, Bacteroidetes, Firmicutes, Spirochaetes and Verrucomicrobia by undigested-carbohydrate fermentation. Thus, fiber amount and type determine their occurrence. FPs bind receptors from the pattern recognition family, those from commensal bacteria induce a different response than those from pathogens. PSA is a capsular polysaccharide from B. fragilis stimulating immunoregulatory protein expression, promoting IL-2, STAT1 and STAT4 gene expression, affecting cytokine production and response modulation. PAs interact with neonatal immunity, contribute to gut maturation, modulate the gut–brain axis and regulate host immunity. SLPs are composed of a sphingoid attached to a fatty acid. Prokaryotic SLPs are mostly found in anaerobes. SLPs are involved in proliferation, apoptosis and immune regulation as signaling molecules. The AhR is a transcription factor regulating development, reproduction and metabolism. AhR binds many ligands due to its promiscuous binding site. It participates in immune tolerance, involving lymphocytes and antigen-presenting cells during early development in exposed humans.

The Role of Leptin in Childhood Immune Thrombocytopenia (ITP): An Anti-Inflammatory Agent?

To investigate the effect of leptin in childhood ITP, we measured plasma leptin in 39 children with acute ITP, after treatment and in remission, and in 33 healthy age/BMI-matched controls. We also cultured ITP and control peripheral blood mononuclear cells (PBMCs) with recombinant leptin to assess its direct effect on pro/anti-inflammatory cytokine gene expression. A significant increase in leptin was observed in children with active disease compared to controls. A significant inverse correlation of leptin with platelet count was also observed in children with acute ITP. Leptin remained high after treatment with IVIg, whereas steroid treatment lowered leptin below control levels. In remission, leptin was in the control range. Cytokine gene expression was significantly increased in children with acute ITP compared with controls, with highest expression for IFN-γ and IL-10. IVIg/steroid treatment significantly decreased IFN-γ and IL-10 expression. In remission, IFN-γ and IL-10 expression remained low. Addition of leptin to PBMCs isolated from patients in remission resulted in a significant increase in IL-10 gene expression compared to controls. Further experiments with purified T-cells and monocytes identified monocytes as the source of leptin-induced IL-10. We suggest that leptin acts as an active anti-inflammatory agent in childhood ITP by promoting IL-10 secretion by monocytes.

Peripheral blood mononuclear cell response to YKL-40 and Galectin-3 in cystic fibrosis

BackgroundElevated circulating levels of YKL-40 correlate with disease severity in Cystic Fibrosis (CF), but the role of YKL-40 in the inflammatory response in CF is still under investigation. Our main goal was to evaluate if YKL-40 can modulate the expression of major cytokines (IL-6, IL-10, IL-13) implicated in the inflammatory response in CF. A secondary goal was to explore the interactions between YKL-40 and other circulating proteins to determine the impacts on cytokine modulation. MethodPeripheral blood mononuclear cells (PBMCs) were isolated from the blood of 83 adult CF patients in stable clinical condition. PBMCs were treated with human YKL-40 followed by the measure of IL-6, IL-10 and IL-13 gene expression. Protein arrays were used to explore the interactions between YKL-40 and circulating proteins. Interaction with Galectin-3 (GAL3) was identified, and confirmed by binding assay. Cytokine gene expressions were again monitored by RT-qPCR after PBMC treatment with GAL3, with or without YKL-40 co-stimulation. ResultsFollowing YKL-40 stimulation, PBMC gene expression of IL-6, IL-10 and IL-13 varies across patients. IL-6 and IL-13 are coexpressed, but this response was different in male and female patients. GAL3 protein was detected in the blood of CF patients, and a molecular interaction with YKL-40 was identified. GAL3 did not interfere with the YKL-40 stimulation of IL-6, IL-10 and IL-13 but may modulate the coexpression. ConclusionWe observed that YKL-40 stimulation had a variable impact on IL-6, IL-10, and IL-13 gene expression in CF PBMCs and uncovered an interaction between GAL3 and YKL-40 in the serum of CF patients. Our findings suggest that YKL-40 is not only a biomarker of disease severity in CF, but it might play an active role in the inflammatory pathophysiology of the disease.

Revisiting bupropion anti-inflammatory action: involvement of the TLR2/TLR4 and JAK2/STAT3.

There are accumulating reports regarding poor response to common antidepressant therapy. Antidepressant resistance is often linked to inflammatory system activation and patients displaying inflammation prior to the treatment are less responsive to antidepressants. We hypothesized that the inefficacy of antidepressant therapy in some patients may be attributable to the drugs' inflammatory mode of action, which has been overlooked because of their substantial therapeutic benefit. Bupropion is a commonly prescribed antidepressant that is often used to treat seasonal affective disorders as well. Nevertheless, research suggests that bupropion causes inflammation and worsens depressive symptoms. Therefore, we investigated the impact of bupropion on cytokines of innate and adaptive immunity, as well as immune signaling pathways. We treated lipopolysaccharide (LPS)-stimulated human peripheral blood mononuclear cells (PBMCs) with different doses of bupropion. Pro-/anti-inflammatory cytokines [tumor necrosis factor alpha (TNFα), interleukin-1β (IL-1β), IL-17, and IL-10] were assessed at both transcriptional and translational levels as well as the involvement of JAK2 /STAT3, TLR2, and TLR4 signaling in this process. Bupropion reduced IL-17A, TNFα, and IL-1β protein levels in the cultures. Nonetheless, bupropion increased IL-1β (P < 0.0001), TNFα (P < 0.0001), and IL-17A (P < 0.05) mRNA levels. Treatment enhanced both IL-10 concentration (P < 0.0001) and gene expression (P < 0.0001). TLR2 (P < 0.0001), TLR4 (P < 0.0001), JAK2 (P < 0.0001), and STAT3 (P < 0.0001) gene expression also rose in response to bupropion. The findings imply that bupropion, particularly 50μM and 100μM, has pro-inflammatory effects and should be co-administered with anti-inflammatory medications, at least in patients with inflammatory conditions.

Effects of co-fermented collagen peptide-jackfruit juice on the immune response and gut microbiota in immunosuppressed mice

Recently, the development of fermented foods for ameliorating immunity and gut microbiota has attracted extensive attention. In this study, co-fermented collagen peptide-jackfruit juice was used in immunosuppressed mice to evaluate the immune response and gut microbiota modulatoration. The results showed that co-fermented peptide-jackfruit juice (FPJ) increased anti-oxidant capacity and lactic acid content. FPJ significantly promoted the immune organ (spleen and thymus) indexes, alleviated the injuries of colon tissue and stimulated the secretion of cytokines and immunoglobulins (IgA, IgM, IgG) (P < 0.05). Moreover, FPJ significantly upregulated the gene expression of TNF-α, IL-4, IFN-γ, IL-2, IL-10, T-bet, Foxp3, RORγ and GATA3. Furthermore, FPJ improved gut microbiota composition and elevated the short-chain fatty acids (SCFAs) concentration. The relative abundances of pathogenic bacteria decreased while beneficial bacteria increased after administration FPJ. These findings suggested that FPJ could be developed as a promising functional food for immunomodulation.

B-Cell Precursor Acute Lymphoblastic Leukemia (BCP-ALL) with t(5;14)(q31;q32)/ IL3-IGH Rearrangement and Eosinophilia: A Peculiar IGH BCP-ALL

Introduction: BCP-ALL associated with t(5;14)(q31;q32) is an exceptional cause of eosinophilia. The IL3 gene expression becomes regulated by the IGH enhancer leading to an overproduction of IL3 causing eosinophilia. Clinical, biological and outcome data are extremely scarce in the literature.Methods: We recruited 2 newly diagnosed patients (pts) with t(5;14)(q31;q32) in our centers. A retrospective search in the French protocol FRALLE 2000 data base and collaboration with the GFCH (Groupe Francophone de Cytogénétique Hématologique) added 4 other pts. Array CGH, MLPA or genomic PCR search for IKZF1 deletion were performed in these 6 pts. We further analysed the 12 pts with t(5;14)(q31;q32) described in the literature with our 6 newly described pts, leading to a meaningful series of 18 pts.Results: Main features at onset are displayed in table 1. Median age at diagnosis is 16.5 years. Male/female ratio is 4.6. Eosinophilia-related symptoms are common with neurologic, thromboembolic or lung involvement (respectively 4/13, 4/13 and 6/13 pts). No pt exhibits blast cells in CSF at diagnosis. Median WBC count is high (48x109/L), mainly due to a high rate of eosinophils (median: 30.5x109/L). Blood eosinophilia preceded ALL diagnosis from 6 and 12 months in 2 cases. Peripheral blast cells are low or undetectable (median: 0x109/L; range: 0-18.7x109/L). A high basophilia is frequent (median: 0.55x109/L). Bone marrow morphology is marked by a low blast infiltration (median: 47%) and increased eosinophil-lineage cells (median: 24%) with dysplastic features in 6/10 pts. All cases are of the B lineage. All 18 pts but one present the t(5;14)(q31;q32) translocation. For the remaining pt, it was revealed by sequencing as karyotype was normal in a context of eosinophilia. Six out of 18 pts have additional cytogenetic abnormalities. For the first time, we report an IKZF1 deletion which is present in 4 out of 6 pts. Few other deletions are found in more than one pt (PAX5, ETV6, MEF2C, CDKN2A). Remission and outcome data are available for 11 pts (our 6 newly described pts and 5 from the literature). Median follow-up (FU) at point date is 18 months (range: 4 days-30 months). Only 5 pts achieve a first complete remission (CR1) at first attempt. The 6 newly described pts are currently alive: 4 are in continuous CR1 (CCR1) including 2 after hematopoietic stem cell transplantation (HSCT) and 2 are in CCR2 (including 1 after HSCT). The 5 pts from the literature died, 1 from cardiac arrest secondary to eosinophilia and the 4 others from disease progression and / or sepsis.Discussion and Conclusion : 1) t(5;14)(q31;q32)-associated BCP-ALL has remarkable features including some which are shared with other IGH BCP-ALL, according to data from a comprehensive UK retrospective study using IGH FISH screening1. Both conditions affect preferentially teenagers and young adults (median age: 16 years1). The highly biased male to female ratio reported here is milder in other IGH BCP-ALL (1.441). In both entities a low blast count (<50 G/L in 119/158 pts1) is observed. Our new data revealed a high rate of IKZF1 deletion in 4/6 pts (66%) also observed in 35/88 pts (40%) in IGH BCP-ALL1. Differences obviously include the eosinophilia and related symptoms but also the low rate of cytopenias possibly linked to the partial bone marrow infiltration, prevailing in t(5;14)(q31;q32)-associated BCP-ALL. 2) In pediatric IGH BCP-ALL pts, 5-year overall survival is 84%1. The lower published survival rate in t(5;14)(q31;q32)-associated BCP-ALL should be balanced by a 100% survival in CR in our 6 newly described pts with obviously a limited sample size and a shorter FU. This may reflect new treatment strategies, including pediatric intensive chemotherapy and MRD-based stratification. HSCT seems to be an option for pts with suboptimal response (induction failure, high MRD) or relapse. 3) Despite being a specific WHO recognized entity, many common clinical and biological features allow to suggest that t(5;14)(q31;q32)-associated BCP-ALL may be considered as a very rare subset of IGH BCP-ALL. Nevertheless depending on its kinetics and severity, the specific eosinophilia can render difficult both diagnosis and management.1. Russell LJ, Enshaei A, Jones L, et al. IGH@ translocations are prevalent in teenagers and young adults with acute lymphoblastic leukemia and are associated with a poor outcome. J. Clin. Oncol. Off. J. Am. Soc. Clin. Oncol. 2014;32(14):1453-1462. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

RUNX1 gene expression changes in the placentas of women smokers.

The placenta can be affected by environmental factors, such as exposure to cigarette smoke. This exposure in the fetal context is considered a risk factor for the development of short-term postnatal diseases, such as asthma. Asthma is an inflammatory disease characterized by predominant acquisition of CD4 T lymphocytes (TLs) of the Th2 type. Transcription factors such as GATA binding protein 3 (GATA3) and STAT6 actively participate in the differentiation of virgin TLs towards the Th2 profile, while transcription factors such as STAT1, T-Box transcription factor 21 (T-BET), RUNX1 and RUNX3 participate in their differentiation towards the Th1 profile. The objective of the current study was to evaluate the impact of exposure to cigarette smoke on the gene expression of STAT1, T-BET, GATA3, IL-4, RUNX1 and RUNX3 during the gestation period, and to determine whether the expression levels of these genes are associated with changes in global methylation. STAT1, GATA3, RUNX1 and RUNX3 protein and mRNA expression levels in the placental tissue of women smokers and non-smoking women were determined via immunohistochemistry and quantitative PCR (qPCR) respectively. Additionally, T-BET and IL-4 mRNA expression levels were determined by qPCR. On the other hand, global methylation was determined via ELISA. In the present study, significant increases were observed in RUNX1 transcription factor expression in placentas from women smokers when compared with placentas of non-smoking women. Similarly, significant increases in the expression of GATA3, IL-4 and RUNX3 mRNA were observed. The changes in gene expression were not associated with changes in the global methylation levels. Finally, a higher frequency of low-birth-weight infants were identified in cases of exposure to cigarette smoke during pregnancy when compared with infants not exposed to cigarette smoke during pregnancy. Thus, the data of the present study contributed to the understanding of the genetic and clinical impacts of exposure to cigarette smoke during pregnancy and its importance in maternal and fetal health.

Leishmanicidal potentials of Gossypium hirsutum extract and its fractions on Leishmania major in a murine model: parasite burden, gene expression, and histopathological profile.

Introduction. Leishmaniasis is a neglected tropical and subtropical disease caused by over 20 protozoan species.Hypothesis. Treatment of this complex disease with traditional synthetic drugs is a major challenge worldwide. Natural constituents are unique candidates for future therapeutic development.Aim. This study aimed to assess the in vivo anti-leishmanial effect of the Gossypium hirsutum extract, and its fractions compared to the standard drug (Glucantime, MA) in a murine model and explore the mechanism of action.Methodology. Footpads of BALB/c mice were infected with stationary phase promastigotes and treated topically and intraperitoneally with G. hirsutum extract, its fractions, or Glucantime, 4 weeks post-infection. The extract and fractions were prepared using the Soxhlet apparatus with chloroform followed by the column procedure.Results. The crude extract significantly decreased the footpad parasite load and lesion size compared to the untreated control group (P<0.05), as revealed by dilution assay, quantitative real-time PCR, and histopathological analyses. The primary mode of action involved an immunomodulatory role towards the Th1 response in the up-regulation of IFN-γ and IL-12 and the suppression of IL-10 gene expression profiling against cutaneous leishmaniasis caused by Leishmania major.Conclusion. This finding suggests that the extract possesses multiple combinatory effects of diverse bioactive phytochemical compositions that exert its mechanisms of action through agonistic-synergistic interactions. The topical extract formulation could be a suitable and unique candidate for future investigation and pharmacological development. Further studies are crucial to evaluate the therapeutic potentials of the extract alone and in combination with conventional drugs using clinical settings.

Non-Thermal Atmospheric Pressure Argon-Sourced Plasma Flux Promotes Wound Healing of Burn Wounds and Burn Wounds with Infection in Mice through the Anti-Inflammatory Macrophages

Plasma medicine is the utilization of gas ionization that might be beneficial for the treatment of burn wounds, a healthcare problem with a significant mortality rate. Due to a lack of information on the impact of plasma flux in immune cells and a high prevalence of bacterial infection in burn wounds, non-thermal argon-based plasma flux was tested on macrophages (RAW246.7) and in mouse models of burn wounds with or without Staphylococcus aureus infection. Accordingly, plasma flux enhanced reactive oxygen species (ROS), using dihydroethidium assay, and decreased abundance of NF-κB-p65 (Western blot analysis) in non-stimulating macrophages. In parallel, plasma flux upregulated IL-10 gene expression (an anti-inflammatory cytokine) in lipopolysaccharide (LPS)-induced inflammatory macrophages, while downregulating the pro-inflammatory cytokines (IL-1β and IL-6). Additionally, plasma flux improved the migratory function of fibroblasts (L929) (fibroblast scratch assay) but not fibroblast proliferation. Moreover, once daily plasma flux administration for 7 days promoted the healing process in burn wounds with or without infection (wound area and wound rank score). Additionally, plasma flux reduced tissue cytokines (TNF-α and IL-6) in burn wounds with infection and promoted collagen in burn wounds without infection. In conclusion, plasma flux induced anti-inflammatory macrophages and promoted the burn-wound healing process partly through the decrease in macrophage NF-κB. Hence, plasma flux treatment should be tested in patients with burn wounds.

Association of −819 T/C IL-10 gene promoter polymorphisms with susceptibility to leprosy in South Sumatera Indonesia

Background: The prevalence of leprosy continues to increase, especially in Indonesia, which is one of the endemic areas of leprosy. One cytokine that plays an important role in the pathogenesis of leprosy is IL-10. Mutations in the gene could cause changes in IL-10 gene expression. One of the most common polymorphisms associated with the IL-10 gene is −819 C/T polymorphism, which is could affect the production of IL-10, and thus influence the process of microbial elimination in the development of leprosy. Objective: To determine the association of −819 C/T interleukin-10 promoter gene polymorphism in leprosy patients treated at RSUP Dr. Mohammad Hoesin Palembang. Method: This study was an analytic observational study with a case–control design. Blood from patients who fulfilled the inclusion criteria was taken through venous puncture and stored in EDTA tubes. DNA isolation was carried out with a DNA blood mini kit (QIAamp) and stored at −20 °C. Polymorphism of −819 C/T interleukin-10 promoter gene was measured by PCR-RFLP method, electrophoresis, and visualized under UV light. Results: In the case group, TT genotype frequency distribution was 21%, CT was 22%, and CC was 7%. In the control group, TT genotype frequency distribution was 28%, CT was 18%, and CC was 4%. In the case group, the allele T frequency distribution was 32% and C allele was 18%. In the control group, the T allele frequency distribution was 37% and C allele was 13%. Conclusion: There is no significant association of −819 C/T Interleukin-10 promoter gene polymorphism with susceptibility to leprosy, both in terms of genotypes and alleles of these genes.