Lower IL-6 Research Articles

Inflammation of Mesenteric Adipose Tissue Correlates with Intestinal Injury and Disease Severity in Rats with Severe Acute Pancreatitis.

Visceral adipose tissue (VAT) is related to SAP prognosis. As a depot of VAT, mesenteric adipose tissue (MAT) resides between pancreas and gut, which might affect SAP and the secondary intestinal injury. To investigate the changes of MAT in SAP. 24 SD rats were randomly divided into four groups. 18 rats in SAP group were euthanized in time gradients (6h, 24h, and 48h after modeling) and the others in control group. Blood samples and tissues of pancreas, gut, and MAT were taken for analysis. Compared to the control group, SAP rats appeared MAT inflammation, presenting higher mRNA expression of TNF-α and IL-6 and lower IL-10, and histological changes after 6h of modeling, which became worse over time. Flow cytometry showed that B lymphocytes increased in MAT after 24h of SAP modeling and lasted up to 48h, earlier than the changes of T lymphocytes and macrophages. The intestinal barrier integrity was damaged after 6h of modeling, presenting lower mRNA and protein expression of ZO-1 and occludin, higher serum levels of LPS and DAO, with pathological changes, which gradually aggravated after 24h and 48h. SAP rats had higher serum levels of inflammatory indicators and revealed histological inflammation of pancreas, the severity of which increased with the passage of modeling time. MAT appeared inflammation in early-stage SAP, and became worse over time, with the same trend as the intestinal barrier injury and the severity of pancreatitis. B lymphocytes infiltrated early in MAT, which might promote the MAT inflammation.

Pro- and Anti-Inflammatory Blood Cytokines Levels in Women with Moderate and Severe Pelvic Venous Insufficiency

The aim of this study was to determine the blood levels of pro- and anti-inflammatory cytokines in patients with moderate and severe pelvic venous insufficiency (PVI). Methods and Results: One hundred and four women with PVI and 30 healthy women (control group [CG]) of reproductive age were examined. The patients with PVI (main group [MG]) were divided into 2 subgroups according to the severity of the disease: MG-moderate (n=63) and MG-severe (n=41). The concentration of pro-inflammatory cytokines (IL-1β, IL-2, IL-6, IL-8), and anti-inflammatory interleukins IL-4 and IL-10 were assessed by enzyme immunoassay using monoclonal antibody panels. Measurements were performed on a microplate photometer. Data analysis showed higher values of pro-inflammatory factors (IL-1β, IL-6, and IL-8) in the MG-moderate than in the CG. The MG-severe, compared with the CG, had high levels of IL-1β, IL-2, IL-6, IL-8, and low IL-10 concentration. In addition, patients of the MG-severe had higher levels of IL-2 than patients of the MG-moderate (P=0.045). The IL-6/IL-10 ratio was characterized by higher values in the MG-moderate and MG-severe than in the CG (P<0.0001). The patients of the MG-severe also had higher levels of the IL-6/IL-10 ratio than patients of the MG-moderate (P=0.020). Conclusion: In patients with PVI with increasing severity of varicose veins, the pro- and anti-inflammatory cytokine balance is progressively disturbed toward the dominance of pro-inflammatory components. The control of these changes in patients is an important component of the design of therapeutic measures and prevention of morphofunctional disorders occurring with the progression of the disease.

Newborns from Mothers Who Intensely Consumed Sucralose during Pregnancy Are Heavier and Exhibit Markers of Metabolic Alteration and Low-Grade Systemic Inflammation: A Cross-Sectional, Prospective Study

Robust data in animals show that sucralose intake during gestation can predispose the offspring to weight gain, metabolic disturbances, and low-grade systemic inflammation; however, concluding information remains elusive in humans. In this cross-sectional, prospective study, we examined the birth weight, glucose and insulin cord blood levels, monocyte subsets, and inflammatory cytokine profile in 292 neonates at term from mothers with light sucralose ingestion (LSI) of less than 60 mg sucralose/week or heavy sucralose intake (HSI) of more than 36 mg sucralose/day during pregnancy. Mothers in the LSI (n = 205) or HSI (n = 87) groups showed no differences in age, pregestational body mass index, blood pressure, and glucose tolerance. Although there were no differences in glucose, infants from HSI mothers displayed significant increases in birth weight and insulin compared to newborns from LSI mothers. Newborns from HSI mothers showed a substantial increase in the percentage of inflammatory nonclassical monocytes compared to neonates from LSI mothers. Umbilical cord tissue of infants from HSI mothers exhibited higher IL-1 beta and TNF-alpha with lower IL-10 expression than that found in newborns from LSI mothers. Present results demonstrate that heavy sucralose ingestion during pregnancy affects neonates' anthropometric, metabolic, and inflammatory features.

Variants of NOD2 in Leishmania guyanensis-infected patients with cutaneous leishmaniasis and correlations with plasma circulating pro-inflammatory cytokines.

Leishmaniases, a group of vector-borne diseases, are caused by the protozoan intracellular parasite Leishmania (L.) and are transmitted by the phlebotomine sandflies. A wide range of clinical manifestations in L- infection is observed. The clinical outcome ranges from asymptomatic, cutaneous leishmaniasis (CL) to severe mucosal leishmaniasis (ML) or visceral leishmaniasis (VL), depending on the L. species. Interestingly, only a fraction of L.-infected individuals progress to disease development, suggesting a key role of host genetics in the clinical outcome. NOD2 plays a critical role in the control of host defense and inflammation. The NOD2-RIK2 pathway is involved in developing a Th1- type response in patients with VL and C57BL/6 mice infected with L. infantum. We investigated whether variants in the NOD2 gene (R702W rs2066844, G908R rs2066845, and L1007fsinsC rs2066847) are associated with susceptibility to CL caused by L. guyanensis (Lg) in 837 patients with Lg-Cl and 797 healthy controls (HC) with no history of leishmaniasis. Both patients and HC are from the same endemic area of the Amazonas state of Brazil. The variants R702W and G908R were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), and L1007fsinsC was by direct nucleotide sequencing. The minor allele frequency (MAF) of L1007fsinsC was 0.5% among the patients with Lg-CL and 0.6% in the healthy controls group. R702W genotypes frequencies were similar in both groups. Only 1% and 1.6% were heterozygous for G908R among the patients with Lg-CL and HC, respectively. None of the variants revealed any association with susceptibility to the development of Lg-CL. Correlations of genotypes with the level of plasma cytokines revealed that individuals with the mutant alleles of R702W tend to have low levels of IFN-γ. G908R heterozygotes also tend to have low IFN-γ, TNF-α, IL-17, and IL-8. Variants of NOD2 are not involved in the pathogenesis of Lg-CL.

Cardiovascular health profiles, systemic inflammation, and physical function in older adults: A population-based study

We examined the association of modifiable cardiovascular health (CVH) metrics with physical function among rural older adults in China and the potential role of inflammatory mechanisms in the association. This study included 3733 stroke- and dementia-free participants (age ≥65 years; 56.9% women) in the baseline survey of a multimodal intervention study in rural China. From March-September 2018, data were collected via face-to-face interviews, clinical assessments, and laboratory tests. The Short Performance Physical Battery (SPPB) test was performed to assess physical function. We defined six modifiable CVH metrics according to the modified American Heart Association's recommendations. Serum interleukin (IL)-6 was measured in a subsample (n = 1156). Data were analyzed with multiple general linear and logistic regression models and structural equation modeling. Poor physical function (SPPB score ≤9) was defined in 1443 participants. Ideal CVH (vs. poor CVH) was associated with multivariable-adjusted odds ratio of 0.60 (95%CI 0.48–0.75) for poor physical function. Ideal CVH was significantly associated with higher scores on balance, chair stand, and walking speed tests (all p < 0.05). Moreover, ideal CVH profile was associated with lower serum IL-6 (multivariable-adjusted β=-0.04; 95% CI -0.06, -0.01). Mediation analysis revealed that serum IL-6 accounted for 14% of the association of CVH with total SPPB score and 10% of the association with walking speed score (p < 0.05). This study suggests that an ideal CVH profile is associated with better physical function among stroke- and dementia-free older adults, partly via inflammatory mechanisms. The preventive implications of these findings warrant further investigation in cohort studies.

Cytokine response in asymptomatic and symptomatic Plasmodium falciparum infections in children in a rural area of south-eastern Gabon.

Plasmodium falciparum is a parasite that causes asymptomatic or symptomatic malaria infections in humans depending on various factors. These infections are also a major cause of anemia in intertropical countries such as Gabon. Past studies have clearly demonstrated that inflammatory markers such as cytokines play a key role in the pathogenesis of malaria disease. However, the clinical manifestations of severe malaria vary according to the level of transmission and more information is needed to gain a better understanding of the factors involved. As such, the objective of this study was to investigate the circulating levels of nine cytokines in asymptomatic and symptomatic P. falciparum infections in Gabonese children and their roles in the pathogenesis of anemia. Blood samples were collected from 241 children aged 3 to 180 months in Lastourville, south-eastern Gabon. Diagnosis of P. falciparum infection was performed using Rapid Diagnosis Tests, microscopy and nested PCR. Levels in the plasma of the Th1 (IFN-γ, TNF-α, IL-6 and IL-12p70), Th17 (IL-17A and IL-22) and Th2 (IL-10, IL-4 and IL-13) cytokines were measured by ELISA. Data showed that IL-6, IFN-γ, IL-12p70, IL-10, and IL-13 levels were significantly higher in children with symptomatic P. falciparum infection compared to uninfected children. IL-10 levels were significantly higher in symptomatic children than in asymptomatic children, who had moderately increased levels compared to uninfected controls. Moreover, only IL-10 and IL-6 levels were significantly higher in children with severe malarial anemia compared to children with uncomplicated malaria who had significantly lower IL-10 levels than children with moderate malarial anemia. These data indicate that the progression of P. falciparum infection towards an advanced stage in children is accompanied by a significant increase in type Th1 and/or Th2 cytokines. These inflammatory mediators could serve as potential predictors of anemia for malaria patients.

Dietary Bacillus licheniformis shapes the foregut microbiota, improving nutrient digestibility and intestinal health in broiler chickens.

Bacillus licheniformis is considered a potential alternative to antibiotic growth promoters of animal growth and health. However, the effects of Bacillus licheniformis on the foregut and hindgut microbiota, and their relationships with nutrient digestion and health, in broiler chickens remain unclear. In this study, we aimed to identify the effects of Bacillus licheniformis BCG on intestinal digestion and absorption, tight junctions, inflammation, and the fore- and hind-gut microbiota. We randomly assigned 240 1-day-old male AA broilers into three treatment groups: CT (basal diet), BCG1 (basal diet + 1.0 × 108 CFU/kg B. licheniformis BCG), and BCG2 (basal diet + 1.0 × 109 CFU/kg B. licheniformis BCG). On day 42, the jejunal and ileal chyme and mucosa were subjected to analysis of digestive enzyme activity, nutrient transporters, tight junctions, and signaling molecules associated with inflammation. The ileal and cecal chyme were subjected to microbiota analysis. Compared with the CT group, the B. licheniformis BCG group showed significantly greater jejunal and ileal α-amylase, maltase, and sucrase activity; moreover, the α-amylase activity in the BCG2 group was higher than that in the BCG1 group (P < 0.05). The transcript abundance of FABP-1 and FATP-1 in the BCG2 group was significantly greater than that in the CT and BCG1 groups, and the GLUT-2 and LAT-1 relative mRNA levels were greater in the BCG2 group than the CT group (P < 0.05). Dietary B. licheniformis BCG resulted in significantly higher ileal occludin, and lower IL-8 and TLR-4 mRNA levels than observed in the CT group (P < 0.05). B. licheniformis BCG supplementation significantly decreased bacterial community richness and diversity in the ileum (P < 0.05). Dietary B. licheniformis BCG shaped the ileac microbiota by increasing the prevalence of f_Sphingomonadaceae, Sphingomonas, and Limosilactobacillus, and contributed to nutrient digestion and absorption; moreover, it enhanced the intestinal barrier by increasing the prevalence of f_Lactobacillaceae, Lactobacillus, and Limosilactobacillus. Dietary B. licheniformis BCG decreased microbial community diversity by diminishing Desulfovibrio, Alistipes, Campylobacter, Vibrio, Streptococcus, and Escherichia coli-Shigella levels, and down-regulating inflammatory associated molecule expression. Therefore, dietary B. licheniformis BCG contributed to digestion and absorption of nutrients, enhanced the intestinal physical barrier, and decreased intestinal inflammation in broilers by decreasing microbial diversity and optimizing the microbiota structure.

Effect of Cancer-Related Cachexia and Associated Changes in Nutritional Status, Inflammatory Status, and Muscle Mass on Immunotherapy Efficacy and Survival in Patients with Advanced Non-Small Cell Lung Cancer

Immune checkpoint inhibitor (ICI)-based immunotherapy has significantly improved the survival of patients with advanced non-small cell lung cancer (NSCLC); however, a significant percentage of patients do not benefit from this approach, and predictive biomarkers are needed. Increasing evidence demonstrates that cachexia, a complex syndrome driven by cancer-related chronic inflammation often encountered in patients with NSCLC, may impair the immune response and ICI efficacy. Herein, we carried out a prospective study aimed at evaluating the prognostic and predictive role of cachexia with the related changes in nutritional, metabolic, and inflammatory parameters (assessed by the multidimensional miniCASCO tool) on the survival and clinical response (i.e., disease control rate) to ICI-based immunotherapy in patients with advanced NSCLC. We included 74 consecutive patients. Upon multivariate regression analysis, we found a negative association between IL-6 levels (odds ratio (OR) = 0.9036; 95%CI = 0.8408-0.9711; p = 0.0025) and the miniCASCO score (OR = 0.9768; 95%CI = 0.9102-0.9999; p = 0.0310) with the clinical response. As for survival outcomes, multivariate COX regression analysis found that IL-6 levels and miniCASCO-based cachexia severity significantly affected PFS (hazard ratio (HR) = 1.0388; 95%CI = 1.0230-1.0548; p < 0.001 and HR = 1.2587; 95%CI = 1.0850-1.4602; p = 0.0024, respectively) and OS (HR = 1.0404; 95%CI = 1.0221-1.0589; p < 0.0001 and HR = 2.3834; 95%CI = 1.1504-4.9378; p = 0.0194, respectively). A comparison of the survival curves by Kaplan-Meier analysis showed a significantly lower OS in patients with cachexia versus those without cachexia (p = 0.0323), as well as higher miniCASCO-based cachexia severity (p = 0.0428), an mGPS of 2 versus those with a lower mGPS (p = 0.0074), and higher IL-6 levels (>6 ng/mL) versus those with lower IL-6 levels (≤6 ng/mL) (p = 0.0120). In conclusion, our study supports the evidence that cachexia, with its related changes in inflammatory, body composition, and nutritional parameters, is a key prognostic and predictive factor for ICIs. Further larger studies are needed to confirm these findings and to explore the potential benefit of counteracting cachexia to improve immunotherapy efficacy.

Evaluation of Host Constitutive and Ex Vivo Coccidioidal Antigen-Stimulated Immune Response in Dogs with Naturally Acquired Coccidioidomycosis.

The early innate immune response to coccidioidomycosis has proven to be pivotal in directing the adaptive immune response and disease outcome in mice and humans but is unexplored in dogs. The objectives of this study were to evaluate the innate immune profile of dogs with coccidioidomycosis and determine if differences exist based on the extent of infection (i.e., pulmonary or disseminated). A total of 28 dogs with coccidioidomycosis (pulmonary, n = 16; disseminated, n = 12) and 10 seronegative healthy controls were enrolled. Immunologic testing was performed immediately, without ex vivo incubation (i.e., constitutive), and after coccidioidal antigen stimulation of whole blood cultures. Whole blood cultures were incubated with a phosphate-buffered solution (PBS) (negative control) or a coccidioidal antigen (rCTS1 (105-310); 10 µg/mL) for 24 h. A validated canine-specific multiplex bead-based assay was used to measure 12 cytokines in plasma and cell culture supernatant. Serum C-reactive protein (CRP) was measured with an ELISA assay. Leukocyte expression of toll-like receptors (TLRs)2 and TLR4 was measured using flow cytometry. Dogs with coccidioidomycosis had higher constitutive plasma keratinocyte chemotactic (KC)-like concentrations (p = 0.02) and serum CRP concentrations compared to controls (p < 0.001). Moreover, dogs with pulmonary coccidioidomycosis had higher serum CRP concentrations than those with dissemination (p = 0.001). Peripheral blood leukocytes from dogs with coccidioidomycosis produced higher concentrations of tumor necrosis factor (TNF)-α (p = 0.0003), interleukin (IL)-6 (p = 0.04), interferon (IFN)-γ (p = 0.03), monocyte chemoattractant protein (MCP)-1 (p = 0.02), IL-10 (p = 0.02), and lower IL-8 (p = 0.003) in supernatants following coccidioidal antigen stimulation when compared to those from control dogs. There was no detectable difference between dogs with pulmonary and disseminated disease. No differences in constitutive or stimulated leukocyte TLR2 and TLR4 expression were found. These results provide information about the constitutive and coccidioidal antigen-specific stimulated immune profile in dogs with naturally acquired coccidioidomycosis.

Anti-inflammatory effects of 2nd generation antipsychotics in patients with schizophrenia: A systematic review and meta-analysis

BackgroundSchizophrenia is a major psychiatric disorder with unknown aetiology. Recent evidence suggests a potential role for cytokines in its pathophysiology and that antipsychotic medication may alter this. While the aetiology of schizophrenia remains only partly understood, an altered immune function representing an important avenue of further discovery. In this systematic review and meta-analysis we focus on the specific effects of second generation antipsychotics risperidone and clozapine on inflammatory cytokines. MethodsA defined systematic search of PubMed and Web of Science databases was performed to identify relevant studies published between Jan 1900 and May 2022. After screening of 2969 papers, 43 studies (27 single-arm and 8 dual-arm) were included that consisted of a total of 1421 patients with schizophrenia in the systematic review. From these, twenty studies (4 dual-arm; 678 patients) had data available on which a meta-analysis could be carried out. ResultsOur meta-analysis showed a significant reduction of pro-inflammatory cytokines post-risperidone treatment in the absence of a similar association with clozapine. Subgroup analyses (First episode v chronic) demonstrated that duration of illness influenced the extent of cytokine alteration; risperidone treatment produced significant cytokine changes (lowered IL-6 and TNF-α) in chronic patients but not in first-episode psychosis (FEP) patients. ConclusionVarying treatment effects on cytokines can be observed by the use of different antipsychotic drugs. The cytokine alterations post-treatment are influenced by the specific antipsychotic drugs and patient status. This may explain disease progression in certain patient groups and influence therapeutic choices in the future.

A phase II biomarker-oriented study to evaluate paclitaxel, olaparib, and durvalumab combination in patients with advanced gastric cancer.

404 Background: DNA damage response (DDR) gene alterations are observed in 27% of gastric cancer (GC), therefore, targeting DDR might be an interesting strategy in GC. Olaparib plus paclitaxel showed numerically improved overall survival (OS) compared to paclitaxel in 2L GC patients (pts), even though it did not meet the endpoint. (Lancet Oncol 2017). Besides DNA damage repair, olaparib induces immune modulation. The role of immune checkpoint inhibitor (ICI) is established in 1L and 3L+ GC. The combination of cytotoxic chemotherapy, DDR acting agent and ICI might have synergistic effects in GC, which has not been tested in clinical studies. Here, we report biomarker-oriented phase II study of paclitaxel/olaparib/durvalumab combination in 2L GC. (NCT03579784). Methods: Advanced GC pts who have failed to 1L chemotherapy were enrolled. During 1st cycle, paclitaxel (80 mg/m2, D1, 8, 15 Q 4wk) and olaparib (150mg bid, D1-28) was administered. From 2nd cycle, durvalumab (1500 mg, D1 Q 4wk) was added on them. Tumor biopsy (pretreatment, after 1st cycle, at disease progression) and blood collection (every cycle) was mandatory. Primary endpoint was the disease control rate (DCR). Secondary endpoints included ORR, DoR, PFS, OS and safety. For biomarker study, multiplex IHC, NGS, ctDNA, and cytokines and angiogenic factors analysis were performed. Efficacy analysis set (ES) and safety analysis set (SS) were defined as pts who underwent at least one tumor assessment and all pts who received cycle1 day1 or more, respectively. Results: Fifty pts were enrolled (6 screening fail or withdrawal of consent, 44 in SS, 38 in ES) HER2+ GC were 13.2%. Among 38 in ES, DCR and ORR were 84.1% (95% CI, 72.5-95.7) and 39.5% (95% CI, 24.0-55.0) (1 CR, 14 PR and 17 SD). PFS, DoR and OS were 6.7 (95% CI, 4.5-10.3), 6.5 (95% CI, 5.1-10.7) and 11.4 (95% CI, 9.2-13.2) months. Homologous recombination repair (HRR) genetic alterations (9.6 vs. 5.9 in PFS, p=0.037), high CD8/CD3 ratio (8.8 vs. 3.7 in PFS, p=0.008), high PD-L1 (CPS≥10) (9.6 vs 5.1 in PFS, p=0.031), low ATM IHC (13.3 vs. 9.8 in OS, p=0.066), and low serum IL-6 (12.9 vs. 8.3 in OS, p=0.001) were associated with prolonged PFS or OS. In SS, most adverse events (AEs) were mild, and Gr 3/4 leukopenia and anemia were observed in 52.2% and 31.8%. There were no treatment related AEs resulted in study discontinuation. Conclusions: Paclitaxel/olaparib/durvalumab is a promising combination in 2L GC, and is worth to be further explored, especially in biomarker-enriched patients with HRR, specific T-cells subsets, high PD-L1, low ATM and IL-6. Clinical trial information: NCT03579784 .

Abstract WP226: Circadian Rhythm Influences Immune Response In Ischemic Stroke Mice

Background: Stroke leads to disability and death worldwide. There is evidence that stroke affects the immune system function, and the clock gene controls the immune system. Stroke elevates the inflammatory cascade. Immune response controls the stroke pathology. However, it is unclear if the circadian rhythm influences the immune system in ischemic stroke mice and affects stroke outcomes. Hypothesis: We hypothesized that immune response might be affected by a circadian rhythm that aggravates stroke pathology in a mouse suture occlusion model Methods: Seven to eight-month-old C57BL/6J (Wild Type, n=8-10 mice/group) mice were randomly assigned to do stroke at the different time points of the day following zeitgeber time at ZT0, ZT6, ZT12, and Z18. Cerebral Ischemia was induced by occlusion of the middle cerebral artery (MCAO) for 60 min. Whole blood was analyzed using flow cytometry, and we observed macrophages (M1, M2), neutrophils (N1, N2), Anti-inflammatory IL10, and pro-inflammatory TNF-α cytokines at 24h and 48h. Forty-eight hours after stroke, TTC staining was done to estimate brain infarction, and the infarct area was measured using NIH-Image J software. Results: There was a significant increase ( p &lt;0.005) in TNF-α (9±2.50) and significant low IL-10 (5.12±1.35) ( p &lt;0001) at (ZT6, noon) stroke at 48h during a deep sleep period (ZT6, noon) stroke in comparison to fully awake period stroke. We found a significant increase ( p &lt;001) in M1 (54.12±5.16) macrophage and a significant decrease ( p &lt;001) in M2 (45.87±5.16) macrophage at ZT6 (noon) compare to other zeitgeber time points. We also found a significantly higher M1:M2 ratio (1.17) at ZT6. Additionally, we found that neutrophil N1 (66.88±4.39) level was significantly ( p &lt;0001) elevated while neutrophil N2 (33.88±4.43) was significantly reduced at ZT6 (noon) sleep period in comparison to ZT0, ZT12, and Z18 time points. We also found a considerably higher N1:N2 ratio (1.97) at ZT6. Conclusion: This study demonstrates that mice brain infarcts are influenced by immune responses that aggravate stroke pathology during their sleep period (noon/ZT6) than during their awake period (midnight/ZT18).

Correlation of high serum interleukin-6 with clinical outcome and T cell response in patients with unresectable hepatocellular carcinoma treated with atezolizumab and bevacizumab.

596 Background: We elucidated the clinical and immunologic implications of serum interleukin (IL)-6 levels in patients with unresectable hepatocellular carcinoma (HCC) treated with atezolizumab and bevacizumab (Ate/Bev). Methods: We prospectively enrolled 165 patients with unresectable HCC (Discovery cohort: 84 patients from three centers; Validation cohort: 81 patients from one center). Baseline blood samples were analyzed using a flow cytometric bead array. The tumor immune microenvironment was analyzed using RNA sequencing. Results: In the Discovery cohort, clinical benefit (CB) was defined as a complete or partial response or stable disease for ≥ 6 months. Among various blood-based biomarkers, the serum IL-6 level was significantly higher in patients without CB than in those with CB (mean 11.56 vs. 5.05 pg/mL, P=0.02). The optimal cutoff value for high IL-6 was determined as 18.49 pg/mL using maximally selected rank statistics, and 15.2% of patients were identified to have high IL-6 levels at baseline. In both the Discovery and Validation cohorts, patients with high baseline IL-6 levels had a reduced response rate and worse progression-free and overall survival after Ate/Bev compared with that of patients with low IL-6 levels. In the multivariable Cox regression analysis, the clinical implication of high IL-6 levels persisted even after adjustment for various confounding factors. Patients with high IL-6 levels showed reduced interferon-γ and tumor necrosis factor-α secretion from CD8+ T cells. Moreover, excess IL-6 suppressed cytokine production and CD8+ T-cell proliferation. Finally, patients with high IL-6 levels exhibited a non-T-cell-inflamed immunosuppressive tumor microenvironment. Conclusions: High baseline IL-6 levels can be associated with poor clinical outcomes and impaired T-cell function in patients with unresectable HCC after Ate/Bev treatment.

Nanoparticles in induced sputum - a window to airway inflammation among active smokers.

Aims: To evaluate the role of nanoparticles (NP) in sputum samples of active smokers as markers of inflammation and disease. Materials & methods: 29 active smokers were included (14 with chronic obstructive pulmonary disease [COPD]) and underwent clinical assessment, pulmonary function tests, sputum induction (with NPanalysis) and blood sampling. Results: Higher particle and NP concentrations and smaller mean size directly correlated with clinical parameters such as the COPD Assessment Test score and impulse oscillometry results. Similar correlations were found between NPs and increased sputum IL-1β, IL-6 and TNF-α. Among COPD patients, higher IL-8 and lower IL-10 serum levels also correlated with NP concentrations. Conclusion: This proof-of-concept study shows the potential of sputum NPs as markers of airway inflammation and disease.

Similar to Those Who Are Breastfed, Infants Fed a Formula Containing 2′-Fucosyllactose Have Lower Inflammatory Cytokines in a Randomized Controlled Trial

Background: Evidence suggests that human milk oligosaccharides (HMOs) provide multiple benefits to infants, including prebiotic effects, gut maturation, antimicrobial activities, and immune modulation. Clinical intervention studies with HMOs are required to confirm these benefits in infants.Objective: Our objective was to investigate the effects of feeding formulas supplemented with the HMO 2′-fucosyllactose (2′-FL) on biomarkers of immune function in healthy term infants.Methods: We performed a substudy nested within a randomized, double-blind, controlled growth and tolerance study in healthy singleton infants (birth weight ≥2490 g) who were enrolled by 5 d of life and exclusively formula-fed (n = 317) or breastfed (n = 107) from enrollment to 4 mo of age. Formula-fed infants were randomly assigned to receive 1 of 3 formulas, all containing 2.4 g total oligosaccharides/L [control: galacto-oligosaccharides (GOS) only; experimental formulas: GOS + 0.2 or 1.0 g 2′-FL/L], and compared with a breastfed reference group. For this substudy, blood samples were drawn from infants at 6 wk of age (n = 31–42/group). Peripheral blood mononuclear cells (PBMCs) were isolated for cellular phenotyping and stimulated ex vivo with phytohemagglutinin for proliferation and cell cycle progression or respiratory syncytial virus (RSV). Cytokine concentrations were measured in plasma and in ex vivo–stimulated culture supernatants.Results: Breastfed infants and infants fed either of the experimental formulas with 2′-FL were not different but had 29–83% lower concentrations of plasma inflammatory cytokines than did infants fed the control formula [interleukin (IL) receptor antagonist (IL-1ra), IL-1α, IL-1β, IL-6, and tumor necrosis factor α (TNF-α)] (P ≤ 0.05). In ex vivo RSV–stimulated PBMC cultures, breastfed infants were not different than either of the groups fed formula with 2′-FL, but they had lower concentrations of TNF-α (31%) and interferon γ (IFN-γ 54%) (P ≤ 0.05) and tended to have lower IL-1ra (25%) and IL-6 (38%) (unadjusted P ≤ 0.05) and IL-1β (30%) (unadjusted P = 0.06) than did infants fed the control formula.Conclusions: Our data indicate that infants fed formula supplemented with 2′-FL exhibit lower plasma and ex vivo inflammatory cytokine profiles, similar to those of a breastfed reference group. This trial was registered at clinicaltrials.gov as NCT01808105.

Lower Creatine Kinase, IL-6 Levels and Femoral Offset Values Lead to Better Outcome Harris Hip Score Post Bipolar Hemiarthroplasty Surgery Day 14 and Day 30

Bipolar Hemiarthroplasty (HA) procedure allows patients after a neck femur fracture to be able to perform early mobilization. Hence complications due to prolonged bed rest could be prevented. Early mobilization should be achieved immediately after bipolar HA surgery. There are several factors affecting early mobilization in patients, some of which include tissue damage during surgery (creatine kinase), inflammatory status of patients (IL-6) and implant position (femoral offset). This study aims to show whether lower levels of creatine kinase, IL-6 and femoral offset can result in better Harris Hip Score following Bipolar HA surgery day 14 and day 30. By knowing the influencing factors, it is hoped that more accurate management could be carried out. Thus, the outcome parameters of early mobilization in the form of returning to daily life and work activities are getting better and the risk of complications can be avoided. This study compared the Harris Hip Score (HHS) in patients who'd already had Bipolar Hemiarthroplasty using a prospective cohort study design (Bipolar HA) who met the inclusion criteria at the Central General Hospital (RSUP) Prof I.G.N.G Ngoerah Denpasar and provided them informed consent. In order to do descriptive analysis, normality and homogeneity tests, and proportion comparison analysis, the research data was entered into a research sheet and processed using a computer and the SPSS for Windows version 26 program. In this study, the sample data (n=32) was not normally distributed. Lower levels of IL-6 (&lt; 15 pg/ml), and FO values ​​(​​&lt; 42.5 mm) resulted in better HHS values ​​in patients following Bipolar HA surgery day-14. Additionally, lower levels of CK (&lt; 75 U/L), IL-6 (&lt; 15 pg/ml), and FO values (​​&lt; 42.5 mm) ​​resulted in better HHS values ​​in patients following bipolar HA surgery on day-30. Chi-Square analysis showed that the results of CK levels day-14 were statistically insignificant (p= 0.072&gt;0.05), while the results of CK levels day-30 were statistically significant (p=0.033&lt;0.05). Moreover, chi-Square analysis showed that the results of day-14 and day-30 were statistically significant on IL-6 levels (p-0.016&lt;0.05 and p= 0.015&lt;0.05), and FO values (p= 0.012&lt;0.05 and p-0.033&lt;0.05). It is concluded that lower levels of IL-6, and FO values resulted in better HHS values in patients following Bipolar HA surgery day-14. Additionally, lower levels of CK, IL-6, and FO values resulted in better HHS values in patients following bipolar HA surgery on day-30.

Perindopril Reduce SARS-COV-2 Spike-ACE2 Binding, ACE2 Overexpression and Cytokine Storm in Adipocyte Cell Infected with SARS-COV-2

Abstract Funding Acknowledgements Type of funding sources: Public Institution(s). Main funding source(s): Ristekdikti Background and Aims The SARS-CoV-2 virus can infect adipocyte cells via ACE2 Receptor thus triggering ACE2 overexpression and cytokine storms which cause lethal complications. Hence, we explore the effect of Perindopril on the expression of ACE2, IL-6, IL-1B TNF-α in adipocyte cultures infected by SARS-CoV-2 spike protein and identify the possible mechanism involved. Materials and Methods Adipocyte culture obtained from a healthy and obese donor was divided into 4 triplicate groups (P0: negative control without treatment; P1: positive control (SARS-CoV-2 spike protein); P2: SARS-CoV-2 spike protein + exposure to Perindopril 0.5 µM); P3: SARS-CoV-2 spike protein + anti-ACE2 antibody 100 µg/mL; and evaluated at 24 and 48 hours. ACE2 expression was evaluated using immunofluorescence. IL-6, TNFα, and IL-1B were evaluated using ELISA. SARS-COV-2 Spike-ACE2 Binding was evaluated using Competitive ELISA. Data analysis was performed using SPSS 25.0 software. Results At first 24 hours of incubation, perindopril treatment has the highest ACE2 expression compared to negative control, positive control and anti-ACE2 antibody (113.52±0.34 ng/mL vs 13.3±0.87 ng/mL, 90.2±2.73 ng/mL, 17.3±0.11 ng/mL, p&amp;lt;0.01), lower ACE-ACE2R binding compared to anti-ACE2 antibody group (169.52±4.07 ng/mL vs 290.71±6.22 ng/mL, p&amp;lt;0.01) and higher IL-6 expression compared to positive control group (64.65±0.12 ng/mL vs 60.08±0.77 ng/mL p&amp;lt;0.01). Interestingly, after 48 hours, perindopril treatment was shown to prevent further increase of ACE2 expression compared to a positive control (47.37±0.76 ng/mL vs 80.31±5.37 ng/mL, p&amp;lt;0.01), higher SARS-COV-2 Spike-ACE2 binding compared to anti-ACE2 antibody group (143.68±3.68 ng/mL vs 103.1±9.49 ng/mL, p&amp;lt;0.01), and lower IL-6 expression compared to the positive control group (42.66±1.94 ng/mL vs 90.93±2.48 ng/mL p&amp;lt;0.01). However, no significant difference in TNFα and IL-1B expression between perindopril treatment and positive control in both 24 and 48 hours. Conclusion This study showed that perindopril reduces cytokine storm by preventing ACE-2 and IL-6 overexpression via an increasing number of SARS-COV-2 Spike-ACE2 competitive binding in adipocyte culture infected with SARS-COV-2 spike protein. A further clinical trial is needed to prove the benefit of perindopril in obese patients with COVID-19.

Low Expression of Calcitriol Level and Interleukin-10 and Hypoxia-inducible Factor-1 Alpha Expression on Placenta

BACKGROUND: Preeclampsia (PE) is the disease of theories and the second leading cause of maternal and perinatal morbidities and mortalities worldwide. These pathological disturbances will induce inflammation process, oxidative stress, and poor subsequent growth on the fetus including 32% of intrauterine growth restriction, 22% of prematurity, and 24% of neonatal sepsis and asphyxia. There are many theories about the mechanism of PE. In the molecularly level, it is suspected that the low level of calcitriol and interleukin (IL)-10 expressions and high expression of hypoxia- inducible factor (HIF)-1α are the risk factors of PE. AIM: The aim of this study was to prove the low calcitriol level, IL-10, and high expression of HIF-1α in the placenta as the risk factors of PE. METHODS: A nested case–control study was conducted at the Department of Obstetrics-Gynecology Sanglah and Wangaya Hospital Denpasar, Bali, from November 2020 to February 2021. RESULTS: A total of 64 samples of 20–40 weeks gestation age were selected by purposive consecutive sampling, divided into two groups consist of 32 PE as the cases and 32 non-PE as the controls. The material examination, 3 × 3 cm was isolated from the maternal placental surface, was performed at Laboratorium Biomedik Terpadu, Faculty of Medicine, Udayana University. We performed an ELISA technique to find out the calcitriol level; in the other hand, we used immunohistochemistry for detected expression of IL-10 and HIF-1-α. The results revealed the risk of PE in low placental calcitriol levels about 13.8 times higher than in high calcitriol levels (odds ratio [OR] = 13,801, 95% confidence interval [CI] = 4.048–47,050, p = 0.001. The risk of PE in low placental IL-10 expression about 6.6 times higher than in high IL-10 expression (OR = 6600; 95% CI = 2208–19,728; p = 0.001). The risk of PE in high placental HIF-1-α expression about 5.6 times higher than in low placental HIF-1-α expression (OR = 5.622; 95% CI = 1.922–16.450; p = 0.001). CONCLUSION: Low calcitriol level, low IL-10, and high HIF-1-α expression on the placenta were proved as significant risk factors for the development of PE.

High serum IL-6 correlates with reduced clinical benefit of atezolizumab and bevacizumab in unresectable hepatocellular carcinoma

We elucidated the clinical and immunologic implications of serum IL-6 levels in patients with unresectable hepatocellular carcinoma (HCC) treated with atezolizumab and bevacizumab (Ate/Bev). We prospectively enrolled 165 patients with unresectable HCC (discovery cohort: 84 patients from three centres; validation cohort: 81 patients from one centre). Baseline blood samples were analysed using a flow cytometric bead array. The tumour immune microenvironment was analysed using RNA sequencing. In the discovery cohort, clinical benefit 6 months (CB6m) was defined as complete or partial response, or stable disease for ≥6 months. Among various blood-based biomarkers, serum IL-6 levels were significantly higher in participants without CB6m than in those with CB6m (mean 11.56 vs. 5.05pg/ml, p= 0.02). Using maximally selected rank statistics, the optimal cut-off value for high IL-6 was determined as 18.49pg/ml, and 15.2% of participants were found to have high IL-6 levels at baseline. In both the discovery and validation cohorts, participants with high baseline IL-6 levels had a reduced response rate and worse progression-free and overall survival after Ate/Bev treatment compared with those with low baseline IL-6 levels. In multivariable Cox regression analysis, the clinical implications of high IL-6 levels persisted, even after adjusting for various confounding factors. Participants with high IL-6 levels showed reduced interferon-γ and tumour necrosis factor-α secretion from CD8+ T cells. Moreover, excess IL-6 suppressed cytokine production and proliferation of CD8+ T cells. Finally, participants with high IL-6 levels exhibited a non-T-cell-inflamed immunosuppressive tumour microenvironment. High baseline IL-6 levels can be associated with poor clinical outcomes and impaired T-cell function in patients with unresectable HCC after Ate/Bev treatment. Although patients with hepatocellular carcinoma who respond to treatment with atezolizumab and bevacizumab exhibit favourable clinical outcomes, a fraction of these still experience primary resistance. We found that high baseline serum levels of IL-6 correlate with poor clinical outcomes and impaired T-cell response in patients with hepatocellular carcinoma treated with atezolizumab and bevacizumab.

Dietary addition of Humulus scandens improved the intestinal barrier in rabbits

ABSTRACT This study was conducted to investigate the effect of Humulus scandens (HS) in diet on the production performance and intestinal barrier of rabbits. One hundred and sixty Laiwu black rabbits at 35 days of age with body weight of 692.5 ± 48.3 g were divided into four groups (five replicates per group and eight rabbits per replicate): fed on a basal diet free of HS (control), 4%, 8% or 12% HS powder supplement. The results showed that the dietary addition of HS decreased the ADFI, F/G, diarrhoea ratio and mortality ratio. Besides, the final body weight and ADG (P < .05) increased with the level of HS. Rabbits in 12% HS group had a higher sIgA and IgG concentration in serum and ileum than control, while lower TNFα, IFN-γ and IL-6 concentration (P < .05). Compared with the control, the HS supplementation groups for 12% level could decrease the mRNA expression of ZO-1, JAM3 and mucin1 (P < .05). In conclusion, dietary supplement of HS modulates immune responses and enhances intestinal barrier, meanwhile inhibits the synthesis of cytokine. Besides, our experiment offers positive evidence in improving rabbit health of HS as rabbit feed resource.