Lower Tumor Necrosis Factor Research Articles

Vitamin D Levels and Markers of Inflammation and Metabolism in HIV-Infected Individuals on Suppressive Antiretroviral Therapy.

Data on vitamin D insufficiency as a cause of inflammation and metabolic dysfunction in HIV-infected individuals are conflicting. We examined the relationships between levels of 25-hydroxyvitamin D [25(OH)D] and biomarkers of inflammation and metabolism in stored blood samples from a prospective trial of vitamin D repletion. Blood samples from HIV-infected individuals on antiretroviral therapy (ART) with HIV-1 RNA <200 copies/ml enrolled in a prospective study were analyzed for 25(OH)D levels, a broad panel of cytokines, highly sensitive C-reactive protein, D-dimer, adiponectin, leptin, and insulin. Correlations between markers and 25(OH)D levels were determined. The Wilcoxon Rank Sum test was used to compare markers between individuals 25(OH)D insufficient and sufficient at baseline and before and after repletion among those who were insufficient and repleted to ≥30 ng/ml after 12 weeks. Of 106 subjects with stored plasma [66 with 25(OH)D <30 ng/ml and 40 ≥ 30 ng/ml], the median age was 50, the CD4 count was 515 cells/mm(3), 94% were male, and the median baseline 25(OH)D was 27 ng/ml. Higher 25(OH)D levels were associated with lower tumor necrosis factor (TNF)-α (r = -0.20, p = 0.04) and higher adiponectin levels (r = 0.30, p = 0.002). Following successful repletion to 25(OH)D ≥30 ng/ml there were no significant changes in inflammatory or metabolic parameters. Our study found associations between low 25(OH)D levels and TNF-α and adiponectin. Repletion did not result in changes in markers of inflammation or metabolism. These data support continued study of the relationship between vitamin D, inflammation, and metabolism in treated HIV infection.

288 Anti-TNF Withdrawal in IBD: Relapse and Recapture Rates and Predictive Factors From 160 Patients in a Pan-UK Study

Introduction The mechanisms behind primary non-response (PNR) to infliximab (IFX) in IBD are still incompletely understood. The role of IFX trough levels (TL) and early antibody formation (ATI) during the induction phase (0-2-6 weeks) are contradicting. Furthermore, the evolution of serum tumor necrosis factor (TNF) during IFX induction has been sparsely studied. Aim We investigated if serum markers of inflammation or drug exposure help in understanding what is driving PNR. Methods In this retrospective single-center study, we identified a cohort of 201 anti-TNF naive Crohn's disease (CD) patients who received IFX induction and had serum samples drawn at weeks 0, 2, 6 and 14. In all samples CRP, albumin, TNF (Prometheus Laboratories Inc.), ATI (measured with drug tolerant homogeneous mobility shift assay, Prometheus Laboratories Inc.) and TL (in-house-developed ELISA) were assessed. PNR was defined as complete absence of clinical improvement at week 14 (physician global assessment). All measurements were compared between responders and those with PNR. Results The incidence of PNR was 8% (n=16). In univariate analysis, low albumin at w6 was associated with PNR (P=0.01, Mann Whitney test). We observed a significant increase of serum TNF after each IFX infusion with medians at w0 and w14 of 1.6 pg/ml (IQR 0.9-2.7) and 7.7 pg/ml (IQR 4.5-11.5) respectively (P<0.0001, KruskalWallis test) (see figure 1). In patients with PNR, this rise in TNF (w14-w0) was significantly lower than in responders (P=0.03, Mann Whitney test). In an attempt to classify inflammation driven by TNF relative to the overall inflammation, we compared TNF/CRP ratio between both groups. A stepwise multiple logistic regression model identified albumin at w6 and TNF/CRP at w0 to be independent significant predictors (P<0.01) of PNR to infliximab at w14, with OR (95% CI) of 0.08 (0.02-0.37) and 3.10 (1.54-6.25) respectively. Conclusions A high disease burden (represented by low albumin, high CRP and serum TNF) and not IFX TL or ATI (detectable in 21% of patients at w14) are the most important factors driving PNR to IFX. TNF and CRP separately did not predict primary response but a higher TNF/ CRP ratio before start of IFX was predictive for PNR, contradicting the theorem that PNR might be due to 'non-TNF-driven' disease. Although the median TL at w14 between both groups did not differ significantly (P=0.48), this ratio indicates that the contribution of TNF in inflammation might even be higher in PNR before IFX start, and possibly demanding a higher loading dose. We further observed an increase of serum TNF after each IFX infusion and this was less in PNR. The mechanism behind this increase remains unclear. These results warrant further investigation for the role of disease burden in PNR to IFX.

286 Drug Concentrations and Antibodies to Infliximab Are Inferior to the Impact of Disease Burden in Primary Non-Response to Infliximab in Crohn's Disease Patients

Introduction The mechanisms behind primary non-response (PNR) to infliximab (IFX) in IBD are still incompletely understood. The role of IFX trough levels (TL) and early antibody formation (ATI) during the induction phase (0-2-6 weeks) are contradicting. Furthermore, the evolution of serum tumor necrosis factor (TNF) during IFX induction has been sparsely studied. Aim We investigated if serum markers of inflammation or drug exposure help in understanding what is driving PNR. Methods In this retrospective single-center study, we identified a cohort of 201 anti-TNF naive Crohn's disease (CD) patients who received IFX induction and had serum samples drawn at weeks 0, 2, 6 and 14. In all samples CRP, albumin, TNF (Prometheus Laboratories Inc.), ATI (measured with drug tolerant homogeneous mobility shift assay, Prometheus Laboratories Inc.) and TL (in-house-developed ELISA) were assessed. PNR was defined as complete absence of clinical improvement at week 14 (physician global assessment). All measurements were compared between responders and those with PNR. Results The incidence of PNR was 8% (n=16). In univariate analysis, low albumin at w6 was associated with PNR (P=0.01, Mann Whitney test). We observed a significant increase of serum TNF after each IFX infusion with medians at w0 and w14 of 1.6 pg/ml (IQR 0.9-2.7) and 7.7 pg/ml (IQR 4.5-11.5) respectively (P<0.0001, KruskalWallis test) (see figure 1). In patients with PNR, this rise in TNF (w14-w0) was significantly lower than in responders (P=0.03, Mann Whitney test). In an attempt to classify inflammation driven by TNF relative to the overall inflammation, we compared TNF/CRP ratio between both groups. A stepwise multiple logistic regression model identified albumin at w6 and TNF/CRP at w0 to be independent significant predictors (P<0.01) of PNR to infliximab at w14, with OR (95% CI) of 0.08 (0.02-0.37) and 3.10 (1.54-6.25) respectively. Conclusions A high disease burden (represented by low albumin, high CRP and serum TNF) and not IFX TL or ATI (detectable in 21% of patients at w14) are the most important factors driving PNR to IFX. TNF and CRP separately did not predict primary response but a higher TNF/ CRP ratio before start of IFX was predictive for PNR, contradicting the theorem that PNR might be due to 'non-TNF-driven' disease. Although the median TL at w14 between both groups did not differ significantly (P=0.48), this ratio indicates that the contribution of TNF in inflammation might even be higher in PNR before IFX start, and possibly demanding a higher loading dose. We further observed an increase of serum TNF after each IFX infusion and this was less in PNR. The mechanism behind this increase remains unclear. These results warrant further investigation for the role of disease burden in PNR to IFX.

Year in review 2014: Lung cancer, pleural diseases, respiratory infections and tuberculosis, bronchoscopic intervention and imaging.

Year in review 2014: Lung cancer, pleural diseases, respiratory infections and tuberculosis, bronchoscopic intervention and imaging.

Species-specific engagement of human nucleotide oligomerization domain 2 (NOD)2 and Toll-like receptor (TLR) signalling upon intracellular bacterial infection: role of Crohn's associated NOD2 gene variants.

Recognition of bacterial peptidoglycan-derived muramyl-dipeptide (MDP) by nucleotide oligomerization domain 2 (NOD2) induces crucial innate immune responses. Most bacteria carry the N-acetylated form of MDP (A-MDP) in their cell membranes, whereas N-glycolyl MDP (G-MDP) is typical for mycobacteria. Experimental murine studies have reported G-MDP to have a greater NOD2-stimulating capacity than A-MDP. As NOD2 polymorphisms are associated with Crohn's disease (CD), a link has been suggested between mycobacterial infections and CD. Thus, the aim was to investigate if NOD2 responses are dependent upon type of MDP and further to determine the role of NOD2 gene variants for the bacterial recognition in CD. The response pattern to A-MDP, G-MDP, Mycobacterium segmatis (expressing mainly G-MDP) and M. segmatisΔnamH (expressing A-MDP), Listeria monocytogenes (LM) (an A-MDP-containing bacteria) and M. avium paratuberculosis (MAP) (a G-MDP-containing bacteria associated with CD) was investigated in human peripheral blood mononuclear cells (PBMCs). A-MDP and M. segmatisΔnamH induced significantly higher tumour necrosis factor (TNF)-α protein levels in healthy wild-type NOD2 PBMCs compared with G-MDP and M. segmatis. NOD2 mutations resulted in a low tumour necrosis factor (TNF)-α protein secretion following stimulation with LM. Contrary to this, TNF-α levels were unchanged upon MAP stimulation regardless of NOD2 genotype and MAP solely activated NOD2- and Toll-like receptor (TLRs)-pathway with an enhanced production of interleukin (IL)-1β and IL-10. In conclusion, the results indicate that CD-associated NOD2 deficiencies might affect the response towards a broader array of commensal and pathogenic bacteria expressing A-MDP, whereas they attenuate the role of mycobacteria in the pathogenesis of CD.

The Galectin-9/Tim-3 pathway is involved in the regulation of NK cell function at the maternal-fetal interface in early pregnancy.

Decidual natural killer (dNK) cells actively participate in the establishment and maintenance of maternal-fetal immune tolerance and act as local guardians against infection. However, how dNK cells maintain the immune balance between tolerance and anti-infection immune responses during pregnancy remains unknown. Here, we demonstrated that the inhibitory molecule T-cell immunoglobulin domain and mucin domain-containing molecule-3 (Tim-3) are expressed on over 60% of dNK cells. Tim-3(+) dNK cells display higher interleukin (IL)-4 and lower tumor necrosis factor (TNF)-α and perforin production. Human trophoblast cells can induce the transformation of peripheral NK cells into a dNK-like phenotype via the secretion of galectin-9 (Gal-9) and the interaction between Gal-9 and Tim-3. In addition, trophoblasts inhibit lipopolysaccharide (LPS)-induced pro-inflammatory cytokine and perforin production by dNK cells, which can be attenuated by Tim-3 neutralizing antibodies. Interestingly, a decreased percentage of Tim-3-expressing dNK cells were observed in human miscarriages and murine abortion-prone models. Moreover, T helper (Th)2-type cytokines were decreased and Th1-type cytokines were increased in Tim-3(+) but not Tim-3(-) dNK cells from human and mouse miscarriages. Therefore, our results suggest that the Gal-9/Tim-3 signal is important for the regulation of dNK cell function, which is beneficial for the maintenance of a normal pregnancy.

Hypertonic saline: a brief overview of hemodynamic response and anti-inflammatory properties in head injury.

Hypertonic saline: a brief overview of hemodynamic response and anti-inflammatory properties in head injury.

Regulatory effect of interleukin-4 in the innate inflammatory response to Rhodococcus aurantiacus infection in mice.

Interleukin (IL)-4 promotes the regression of granulomas during the late phase of Rhodococcus aurantiacus infection. In this study, the contribution of IL-4 to the initial response against this bacterium was investigated using IL-4-deficient mice. Compared with wild-type (WT) mice, IL-4-deficient mice displayed remarkably lower tumor necrosis factor (TNF)-α and IL-6 secretion in the liver, spleen, and blood at the initial phase of infection, along with improved survival. IL-4-deficient mice also showed diminished IL-10 secretion in the spleen and blood; however, hepatic IL-10 levels were similar to those observed in WT animals, and were concomitant with augmented interferon (IFN)-γ production and decreased bacterial burden in the liver at the early infection phase. Histological examination revealed reduced hepatic granuloma formation in infected IL-4-deficient mice. On challenge with heat-killed R. aurantiacus, IL-4-deficient mouse macrophages showed reduced expression of TNF-α, IL-6, and IL-10 at both the gene and protein levels compared with WT mouse cells. These findings indicate that during the initiation of R. aurantiacus-induced inflammation, IL-4 deficiency attenuates cytokine responses in macrophages, which contributes to amelioration in mouse survival and reduction of granulomatous inflammation, and augments a hepatic IFN-γ response which transiently accelerates bacterial elimination.

Perioperative blood transfusion is associated with a gene transcription profile characteristic of immunosuppression: a prospective cohort study

IntroductionBlood transfusion in the perioperative period has frequently been associated with an excess of nosocomial infections. Whilst transfused whole blood induces specific host immune alteration that may predispose to nosocomial infections, the immunomodulating properties associated with leukodepleted blood remain incompletely understood. In this study, we explore the hypothesis that the transfusion of leukodepleted allogeneic blood during or following major gastrointestinal surgery is associated with an immunosuppressed phenotype, which may in turn predispose to postoperative infectious complications.MethodsPatients aged over 45 years undergoing scheduled inpatient major gastrointestinal surgery were recruited. Gene expression profiles of specific inflammatory genes were assayed from blood collected preoperatively, at 24 and at 48 hours after surgery. Genes were selected based on their ability to represent specific immune pathways. Gene expression was quantified using quantitative real-time polymerase chain reaction (qRT-PCR) to measure messenger RNA (mRNA) levels. Postoperative infections were documented using predefined criteria.ResultsOne hundred and nineteen patients were recruited. Fifteen (13%) patients required blood transfusion within 24 hours of surgery, 44 (37%) patients developed infections and 3 (2%) patients died prior to discharge. Patients receiving a blood transfusion were more likely to develop postoperative infections (P =0.02) and to have lower tumour necrosis factor alpha (TNFα), interleukin (IL)-12, IL-23 and RAR-related orphan receptor gamma T (RORγt) gene expression in the postoperative period (P <0.05). The TNFα/IL-10 mRNA ratio at 24 hours (P =0.0006) and at 48 hours (P =0.01) was lower in patients receiving a blood transfusion over this period. Multivariable analysis confirmed that these observations were independent of the severity of the surgical insult.ConclusionsAn association between an immunosuppressive pattern of gene expression and blood transfusion following major elective gastrointestinal surgery is described. This gene expression profile includes a reduction in the activity of innate immunity and T helper cell type 1 (Th1) and T helper cell type 17 (Th17) pathways in those patients receiving a blood transfusion. Blood transfusion was also associated with an excess of infectious complications in this cohort. A mechanistic link is suggested but not proven.Electronic supplementary materialThe online version of this article (doi:10.1186/s13054-014-0541-x) contains supplementary material, which is available to authorized users.

Inhaled corticosteroids and systemic inflammatory response in community-acquired pneumonia: a prospective clinical study.

The previous use of inhaled corticosteroids (ICS) may reduce the inflammatory response and mortality in patients with community-acquired pneumonia (CAP). We measured serum levels of several inflammatory biomarkers, as well as mortality at various time-points, in 663 consecutive patients hospitalized for CAP; 128 (19%) were receiving chronic outpatient treatment with ICS. Patients on previous oral corticosteroids were excluded from the analysis. On admission, patients treated with ICS were older; had been diagnosed with chronic obstructive pulmonary disease (COPD), asthma and pneumonia in the previous year more often; and had higher CAP severity risk classes and lower tumour necrosis factor (TNF)-alpha (P < 0.001) and interleukin (IL)-6 (P = 0.015) serum levels. After adjusting for potential confounders, this association persisted for TNF-alpha (P < 0.001), but not for IL-6. Mortality at 30 and 90 days tended to be lower in patients treated with ICS (P = 0.062 and 0.050, respectively), but mortality was similar after 1 year in both groups (16, 13% vs 81, 15% for patients treated and not treated with ICS, respectively). Hospital readmission rate after 1 year was higher in patients treated with ICS (49, 38% vs 109, 20%, P < 0.001). The association of ICS treatment with a previous diagnosis of pneumonia, lower levels of TNF-alpha and IL-6 on admission and higher readmission rates during follow up persisted in the subpopulation of 210 patients with COPD. Previous use of ICS in patients hospitalized for CAP is associated with a reduced systemic inflammatory response without any impact on long-term mortality.

P090 Direct effect of infliximab on intestinal mucosa sustains mucosal healing: Exploring new mechanisms of action

a b s t r a c t Background: Infliximab is effective in inflammatory bowel disease through several mechanisms, possibly acting at the mucosal level. Aim: To assess the role of infliximab on intestinal mucosa and whether it contributes to mucosal healing. Methods: Human colonic mucosal biopsies were incubated with or without infliximab. Cultured biopsies were evaluated for histological staining, CD68, CD3, E-cadherin and phospho-extracellular signal- regulated kinases (ERK) expression, and apoptosis. A scratch assay and MTT assay were performed with Caco2 cells in the presence of infliximab and/or tumour necrosis factor (TNF)- or treated with super- natants obtained from human peripheral blood mononuclear cells or human intestinal fibroblasts treated with TNF- and infliximab alone or in association. Results: Infliximab-treated biopsies displayed a better histological appearance, reduced inflammation with an increase of E-cadherin, phospho-ERK and apoptosis. Supernatants showed lower TNF-, IL- 17, IL-6 and IL-8 concentration, with an increase in fibroblast-growth-factor. Motility at scratch assay and proliferation at MTT assay of Caco2 cells displayed differential modulation by TNF- and inflix- imab, directly or through supernatants of human intestinal fibroblasts and human peripheral blood mononuclear cells exposed to them. Conclusion: Infliximab contributes to the mucosal healing process by acting directly at an intestinal mucosal level; infliximab indirectly affects epithelial cell migration and proliferation by acting on both fibroblasts and leukocytes.

Perioperative blood transfusion is associated with a gene transcription profile characteristic of immunosuppression: a prospective cohort study

Blood transfusion in the perioperative period has frequently been associated with an excess of nosocomial infections. Whilst transfused whole blood induces specific host immune alteration that may predispose to nosocomial infections, the immunomodulating properties associated with leukodepleted blood remain incompletely understood. In this study, we explore the hypothesis that the transfusion of leukodepleted allogeneic blood during or following major gastrointestinal surgery is associated with an immunosuppressed phenotype, which may in turn predispose to postoperative infectious complications. Patients aged over 45 years undergoing scheduled inpatient major gastrointestinal surgery were recruited. Gene expression profiles of specific inflammatory genes were assayed from blood collected preoperatively, at 24 and at 48 hours after surgery. Genes were selected based on their ability to represent specific immune pathways. Gene expression was quantified using quantitative real-time polymerase chain reaction (qRT-PCR) to measure messenger RNA (mRNA) levels. Postoperative infections were documented using predefined criteria. One hundred and nineteen patients were recruited. Fifteen (13%) patients required blood transfusion within 24 hours of surgery, 44 (37%) patients developed infections and 3 (2%) patients died prior to discharge. Patients receiving a blood transfusion were more likely to develop postoperative infections (P =0.02) and to have lower tumour necrosis factor alpha (TNFα), interleukin (IL)-12, IL-23 and RAR-related orphan receptor gamma T (RORγt) gene expression in the postoperative period (P <0.05). The TNFα/IL-10 mRNA ratio at 24 hours (P =0.0006) and at 48 hours (P =0.01) was lower in patients receiving a blood transfusion over this period. Multivariable analysis confirmed that these observations were independent of the severity of the surgical insult. An association between an immunosuppressive pattern of gene expression and blood transfusion following major elective gastrointestinal surgery is described. This gene expression profile includes a reduction in the activity of innate immunity and T helper cell type 1 (Th1) and T helper cell type 17 (Th17) pathways in those patients receiving a blood transfusion. Blood transfusion was also associated with an excess of infectious complications in this cohort. A mechanistic link is suggested but not proven.

The Respiratory Syncytial Virus Fusion Protein and Neutrophils Mediate the Airway Mucin Response to Pathogenic Respiratory Syncytial Virus Infection

Respiratory syncytial virus (RSV) is the leading cause of death due to a viral etiology in infants. RSV disease is characterized by epithelial desquamation, neutrophilic bronchiolitis and pneumonia, and obstructive pulmonary mucus. It has been shown that infection of BALB/cJ mice with RSV clinical isolate A2001/2-20 (2-20) results in a higher early viral load, greater airway necrosis, and higher levels of interleukin-13 (IL-13) and airway mucin expression than infection with RSV laboratory strain A2. We hypothesized that the fusion (F) protein of RSV 2-20 is a mucus-inducing viral factor. In vitro, the fusion activity of 2-20 F but not that of A2 F was enhanced by expression of RSV G. We generated a recombinant F-chimeric RSV by replacing the F gene of A2 with the F gene of 2-20, generating A2-2-20F. Similar to the results obtained with the parent 2-20 strain, infection of BALB/cJ mice with A2-2-20F resulted in a higher early viral load and higher levels of subsequent pulmonary mucin expression than infection with the A2 strain. A2-2-20F infection induced greater necrotic airway damage and neutrophil infiltration than A2 infection. We hypothesized that the neutrophil response to A2-2-20F infection is involved in mucin expression. Antibody-mediated depletion of neutrophils in RSV-infected mice resulted in lower tumor necrosis factor alpha levels, fewer IL-13-expressing CD4 T cells, and less airway mucin production in the lung. Our data are consistent with a model in which the F and attachment (G) glycoprotein functional interaction leads to enhanced fusion and F is a key factor in airway epithelium infection, pathogenesis, and subsequent airway mucin expression.

Phosphoinositide 3-kinase/Akt pathway is involved in mediating the anti-inflammation effects of magnesium sulfate

BackgroundWe sought to elucidate whether enhancing phosphoinositide 3-kinase (PI3K)/Akt activity is a crucial mechanism underlying the anti-inflammation effects of magnesium sulfate (MgSO4). Materials and methodsMurine macrophages (RAW264.7 cells) were stimulated with endotoxin, endotoxin plus MgSO4, or endotoxin plus MgSO4 plus PI3K inhibitor (LY294002 or wortmannin). Control groups were run simultaneously. After harvesting, we assayed the expression of inflammatory mediators, transcriptional factor nuclear factor κB (NF-κB), and Akt. ResultsMgSO4 significantly attenuated endotoxin-induced upregulation of inflammatory mediators and NF-κB, as macrophages treated with endotoxin plus MgSO4 had significantly lower tumor necrosis factor α (P = 0.022), macrophage inflammatory protein 2 (P = 0.040), phosphorylated (p)-NF-κB p65 (P = 0.003) and p-inhibitor-κB (P < 0.001) concentrations as well as lower NF-κB DNA binding (P = 0.001) than macrophages treated with endotoxin alone. Moreover, macrophages treated with endotoxin plus MgSO4 plus LY294002 or wortmannin had significantly higher tumor necrosis factor α (P = 0.013 and P < 0.001), macrophage inflammatory protein 2 (P = 0.023 and P = 0.003), p-NF-κB p65 (P = 0.006 and P < 0.001), and p-inhibitor-κB (P = 0.001 and P < 0.001) concentrations, as well as higher NF-κB DNA binding (P = 0.038 and P = 0.009), than macrophages treated with endotoxin plus MgSO4, suggesting that PI3K inhibitors reversed these effects of MgSO4. In contrast, macrophages treated with endotoxin plus MgSO4 had significantly higher p-Akt concentration than macrophages treated with endotoxin alone (P = 0.004). Moreover, macrophages treated with endotoxin plus MgSO4 also had significantly higher p-Akt concentration than macrophages treated with endotoxin plus MgSO4 plus LY294002 or wortmannin (P = 0.004 and P < 0.001). ConclusionsEnhancing PI3K/Akt activity is a crucial mechanism underlying the anti-inflammation effects of MgSO4.

Pomegranate Polyphenols and Extract Inhibit Nuclear Factor of Activated T-Cell Activity and Microglial Activation In Vitro and in a Transgenic Mouse Model of Alzheimer Disease

AbstractAlzheimer disease (AD) brain is characterized by extracellular plaques of amyloid β (Aβ) peptide with reactive microglia. This study aimed to determine whether a dietary intervention could attenuate microgliosis. Memory was assessed in 12-mo-old male amyloid precursor protein/presenilin 1 (APP/PS1) transgenic mice via Barnes maze testing followed by division into either a control-fed group provided free access to normal chow and water or a treatment group provided free access to normal chow and drinking water supplemented with pomegranate extract (6.25 mL/L) for 3 mo followed by repeat Barnes maze testing for both groups. Three months of pomegranate feeding decreased the path length to escape of mice compared with their initial 12-mo values (P < 0.05) and their control-fed counterparts (P < 0.05). Brains of the 3-mo study pomegranate-fed mice had lower tumor necrosis factor α (TNF-α) concentrations (P < 0.05) and lower nuclear factor of activated T-cell (NFAT) transcriptional activity (P < 0.05) compared with controls. Brains of the 3-mo pomegranate or control mice were also compared with an additional control group of 12-mo-old mice for histologic analysis. Immunocytochemistry showed that pomegranate- but not control-fed mice had attenuated microgliosis (P < 0.05) and Aβ plaque deposition (P < 0.05) compared with 12-mo-old mice. An additional behavioral study again used 12-mo-old male APP/PS1 mice tested by T-maze followed by division into a control group provided with free access to normal chow and sugar supplemented drinking water or a treatment group provided with normal chow and pomegranate extract–supplemented drinking water (6.25 mL/L) for 1 mo followed by repeat T-maze testing in both groups. One month of pomegranate feeding increased spontaneous alternations versus control-fed mice (P < 0.05). Cell culture experiments verified that 2 polyphenol components of pomegranate extract, punicalagin and ellagic acid, attenuated NFAT activity in a reporter cell line (P < 0.05) and decreased Aβ-stimulated TNF-α secretion by murine microglia (P < 0.05). These data indicate that dietary pomegranate produces brain antiinflammatory effects that may attenuate AD progression.

Antiphotoaging Effects of Light-Emitting Diode Irradiation on Narrow-Band Ultraviolet B–Exposed Cultured Human Skin Cells

Antiaging effects of light-emitting diodes (LEDs) have been clinically demonstrated using one or two wavelengths. The mechanism is unclear. To examine direct and indirect photobiomodulation effects of LEDs on narrow-band ultraviolet B (NB-UVB)-induced photoaging using seven different wavelengths alone or in combination. Four LED wavelengths were chosen based on type I collagen and metalloproteinase (MMP)-1 expression. NB-UVB-irradiated fibroblasts or keratinocytes were irradiated using these four wavelengths. The expression of collagen and MMP-1 in fibroblasts with or without conditioned medium from LED-irradiated keratinocytes and the expression of proinflammatory cytokines in the LED-irradiated keratinocytes were examined. Irradiation with four wavelengths (630, 660, 830, and 850nm) significantly increased the number of viable fibroblasts. These four wavelengths also increased type I collagen expression, particularly four combinations (630/830, 660/850, 630/850, and 660/830nm). The fibroblasts cultured with the keratinocyte conditioned medium, particularly with a combination of 630/850 or 660/830nm, increased collagen levels. Low tumor necrosis factor alpha (TNF-α) and high transforming growth factor beta 1 (TGF-β1) expression was detected in the LED-irradiated keratinocytes. The combination of 630/850- or 660/830-nm irradiation led to better direct and indirect antiphotoaging outcomes than single LED wavelengths in NB-UVB-irradiated cultured normal human skin cells.

Immune Status in Very Preterm Neonates

Preterm neonates are at increased risk of sepsis compared with those born at term. We investigated immune status at birth and early neonatal life in very preterm neonates and its association with short-term outcomes. Prospective observational study conducted at a university hospital recruiting 113 preterm neonates (23-32 weeks) and 78 controls. Monocyte major histocompatibility complex (MHC) class II expression, serum, and ex vivo lipopolysaccharide stimulated levels of six cytokines (tumor necrosis factor α, interleukin (IL)-1β, IL-6, IL-8, IL-10, and IL-12p70) were measured in umbilical cord blood and over the first 7 days. The presence of neonatal sepsis and histologic chorioamnionitis was recorded. Prematurity (preterm labor and preterm premature rupture of membranes cohorts), neonatal sepsis, and histologic chorioamnionitis were associated with significant reduction in monocyte MHC class II expression. Neonates who had evidence of subsequent protracted sepsis had low levels of MHC class II expression at birth. Serial monocyte MHC class II expression revealed a fall by day 2, in all preterm neonates, with the degree being influenced by both prematurity and sepsis, and incomplete recovery by day 7, suggesting immunoparalysis in preterm premature rupture of membranes and preterm labor cohorts. Whole blood lipopolysaccharide stimulation assay showed significantly lower tumor necrosis factor α, values in preterm neonates who subsequently developed sepsis indicating a degree of immunoparalysis. Our data support the concept that fetal exposure to inflammation before preterm delivery leads to subsequent endotoxin hyporesponsiveness (immunoparalysis), which increases the risk of subsequent sepsis and associated organ dysfunction.

Photodynamic Therapy Reduces Bone Resorption and Decreases Inflammatory Response in an Experimental Rat Periodontal Disease Model

The purpose of the present study was to analyze inflammatory conditions after application of photodynamic therapy (PDT) in the gingival tissues of Wistar rats with ligature-induced periodontal disease. The work investigated the effectiveness of PDT in decreasing the inflammatory response, in order to avoid bone loss. Ligatures were positioned at the mandibular first molar of rats (n=6). After 7 days the ligatures were removed and the animals were divided into two groups, one of which received eosin, with both groups subsequently being treated by light-emitting diode irradiation. The animals were killed 7 days after the treatments, and the mandibles were histologically processed (hematoxylin and eosin stain) to assess bone loss, while gingival tissues were removed for quantification of neutrophil infiltration (using the myeloperoxidase assay) and tumor necrosis factor (TNF)-? expression (by ELISA). Histomorphological analysis of periodontal tissues demonstrated that PDT-treated animals presented decreased bone resorption, as well as reduced neutrophil migration and lower TNF-? expression, compared to ligatured animals treated with eosin alone. Within the limitations of this study, it can be concluded that PDT may be useful in the treatment of periodontal disease, because of immunomodulatory effects that decrease the inflammatory response and consequently the bone resorption.

Expression of the Mycobacterium tuberculosis PPE37 protein in Mycobacterium smegmatis induces low tumour necrosis factor alpha and interleukin 6 production in murine macrophages

PPE37 is a member of the Mycobacterium tuberculosis proline-proline-glutamic acid (PPE) multigene family. Its expression is upregulated in bacteria that are phagocytosed by macrophages and is enhanced even more in bacteria isolated from the lungs of infected mice. This raises the possibility that PPE37 may play a role in the virulence of M. tuberculosis and led to this investigation of the function of PPE37. Recombinant bacterial strains, one expressing the M. tuberculosis PPE37 protein (Ms_ppe37) and another harbouring the vector alone (Ms_vec) were generated from the non-pathogenic Mycobacterium smegmatis. These bacterial strains were used to infect peritoneal exudate and bone marrow-derived macrophages. It was found that, despite the comparable intracellular survival between the two recombinant M. smegmatis strains, Ms_ppe37 induced a significantly lower level of tumour necrosis factor alpha and interleukin 6 in the infected macrophages compared with Ms_vec. Western blot analyses revealed that the activation levels of nuclear factor kappa B, mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase and MAPK/p38 were lower in macrophages infected with Ms_ppe37 than in macrophages infected with Ms_vec. These results suggest that PPE37 may have a potential role in interfering with the pro-inflammatory cytokine response of infected macrophages.

Interferon regulatory factor 3 and type I interferons are protective in alcoholic liver injury in mice by way of crosstalk of parenchymal and myeloid cells

Alcoholic liver disease (ALD) features increased hepatic exposure to bacterial lipopolysaccharide (LPS). Toll-like receptor-4 (TLR4) recognizes LPS and activates signaling pathways depending on MyD88 or TRIF adaptors. We previously showed that MyD88 is dispensable in ALD. TLR4 induces Type I interferons (IFNs) in an MyD88-independent manner that involves interferon regulatory factor-3 (IRF3). We fed alcohol or control diets to wild-type (WT) and IRF3 knock-out (KO) mice, and to mice with selective IRF3 deficiency in liver parenchymal and bone marrow-derived cells. Whole-body IRF3-KO mice were protected from alcohol-induced liver injury, steatosis, and inflammation. In contrast to WT or bone marrow-specific IRF3-KO mice, deficiency of IRF3 only in parenchymal cells aggravated alcohol-induced liver injury, associated with increased proinflammatory cytokines, lower antiinflammatory cytokine interleukin 10 (IL-10), and lower Type I IFNs compared to WT mice. Coculture of WT primary murine hepatocytes with liver mononuclear cells (LMNC) resulted in higher LPS-induced IL-10 and IFN-β, and lower tumor necrosis factor alpha (TNF-α) levels compared to LMNC alone. Type I IFN was important because cocultures of hepatocytes with LMNC from Type I IFN receptor KO mice showed attenuated IL-10 levels compared to control cocultures from WT mice. We further identified that Type I IFNs potentiated LPS-induced IL-10 and inhibited inflammatory cytokine production in both murine macrophages and human leukocytes, indicating preserved cross-species effects. These findings suggest that liver parenchymal cells are the dominant source of Type I IFN in a TLR4/IRF3-dependent manner. Further, parenchymal cell-derived Type I IFNs increase antiinflammatory and suppress proinflammatory cytokines production by LMNC in paracrine manner. Our results indicate that IRF3 activation in parenchymal cells and resulting type I IFNs have protective effects in ALD by way of modulation of inflammatory functions in macrophages. These results suggest potential therapeutic targets in ALD.