Higher Levels Of TNF-alpha Research Articles

Defective mTORC1/4E-BP1 signaling contributes to the age-associated reduction of p85 alpha expression in pancreatic acinar cells

Campbell, Katherine E. MD; Song, Jun MS; Starr, Marlene E. PhD; Saito, Hiroshi PhD; Li, Jing PhD; Evers, Mark B. MD, FACS Author Information

SAT0080 Early Rheumatoid Arthritis Patients without Treatment: Baseline Asessment of Insulin Resistance and Cytokines

ObjectivesTo analyze insulin resistance (IR), inflammatory cytokines, adipokines and clinical and laboratory characteristics in patients with early rheumatoid arthritis (ERA) who have not received any treatment for their disease.MethodsThis is...

Cytokines as a predictor of progression to valvular disease in children with rheumatic fever

The immunologic basis of rheumatic fever is well established. However the role of penicillin in the control of the rheumatic process and the prevention of development of rheumatic heart disease is poorly understood. the objective of this study was to monitor the changes in serum levels of Tumour Necrosis Factor (TNF-alpha) and interleukin-8 (IL-8) in children presenting with various stages of rheumatic fever over a time. Study included 42 children aged 6–15 years with RHD followed up over one year by Doppler-echocardiography and laboratory tests to monitor IL-8 and TNF-alpha by ELISA technique. Twenty five children presented with acute arthritis with or without carditis (59.5%), all had statistically significantly high levels of IL-8 and TNF-alpha throughout the follow-up period. Of these 9 (36%) developed rheumatic reactivity and 6 (24%) developed valvular heart disease. Eight children (19%) presented with rheumatic chorea, all had significantly high levels of IL-8 and TNF-alpha throughout the follow-up period, of whom 4 (50%) developed valvular disease. Nine children (21.4%) presented with varying degrees of established chronic rheumatic heart disease, all of whom had no rise in the serum levels of IL-8 and TNF-alpha. These findings indicate that the clinical and epidemiological pattern of rheumatic fever is changing. Immunemodulatory responses could assist us in tracking these changing patterns of disease and assessing current protocols of management.

ADIPONECTIN AND TUMOR NECROSIS FACTOR ALPHA LEVELS, AND THEIR CORRELATIONS WITH ENDOTHELIAL DYSFUNCTION IN CENTRAL OBESITY

Abstract: Background: In obesity, macrophages that infiltrate into adipose tissues create an inflammatory condition. Besides that, adipose tissues release proinflammatory cytokines such as TNF alpha. These cytokines promote LDL oxydation by ROS, and accelerate NO degradation resulting in endothelial dysfunction. Adiponectin which could prevent endothelial dysfunction is decreased secondarily to TNF alpha’s action. Objectives: To determine the serum levels of TNF alpha and adiponectin, and their correlations with the endothelial dysfunction. Methods: This was an observational, descriptive, and analytic study with cross sectional. Samples were students of senior high schools in Manado. Sampling method was carried out consecutively until the required number was enough. Data consisted of age, blood pressure, waist circumference, body weight, lipid profile, creatinin, TNF alpha, adiponectin, and albumin creatinine ratio. We used the Spearman Correlation to analyze the data. Result: Over a 4-month period, there were 35 obese male subjects with ages of 16-18 years old. The average of body weights was 83.23 kg, and of waist circumferences was 103.94 cm. Endothelial dysfunction was found in 5.72% of samples, high levels of TNF alpha in 68.57% of samples, and low levels of adiponectin in 62.8% of samples. We found a significant positive correlation between TNF alpha and endothelial dysfunction (r=0.554, p<0.000), and an inverse correlation between adiponectin level and endothelial dysfunction (r = -0.349, p=0.020). Conclusion: There were high levels of TNF alpha in 68.57% of samples and low levels of adiponectin in 62.86% of samples. There was a significant correlation between TNF alpha level and endothelial dysfunction. There was an inverse correlation between adiponectin level and endothelial dysfunction. Key words: obesity, adiponectin TNF alpha, endothelial dysfunction.

Association of high plasma TNF-alpha levels and TNF-alpha/IL-10 ratios with TNF2 allele in severe P. falciparum malaria patients in Sri Lanka

Plasma levels of pro- and anti-inflammatory cytokines of Plasmodium falciparum-infected patients with severe malaria (SM; n = 62) and uncomplicated malaria (UM; n = 69) from Sri Lanka were assessed. SM patients had significantly higher levels of TNF-alpha (P<0·01), IL-6 (P<0·01), and IL-10 (P<0·05) compared to the UM patients. Plasma IL-2 levels of these patients were undetectable. TNF-alpha levels of a third group of patients with uncomplicated P. falciparum malaria, who were recruited during their fever episodes (UMF; n = 14) were significantly higher than those of the UM patients (P<0·001) and comparable to SM patients. Plasma IFN-gamma levels of SM patients were higher compared to UM patients, but was not statistically significant. Body temperature in both SM and UMF groups were significantly higher compared to UM group, whereas percentages of parasitemia in all three groups were comparable. Analysis of plasma TNF-alpha levels and the ratio of TNF-alpha/IL-10 in UM (n = 34) and SM (n = 34) patients carrying TNF1 and TNF2 allelic types showed that SM patients carrying TNF2 had significantly higher TNF-alpha levels as well as TNF-alpha/IL-10 ratio compared to UM patients carrying TNF1, UM patients carrying TNF2 and SM patients carrying TNF1 (P<0·05). These results suggest that the high circulating TNF-alpha levels and the inadequate IL-10 response in the SM patients carrying TNF2 allele could have contributed to the development of severe falciparum malarial disease.

The effects of a selective inhibitor of c-Fos/activator protein-1 on endotoxin-induced acute kidney injury in mice

BackgroundSepsis has been identified as the most common cause of acute kidney injury (AKI) in intensive care units. Lipopolysaccharide (LPS) induces the production of several proinflammatory cytokines including tumor necrosis factor (TNF)-alpha, a major pathogenetic factor in septic AKI. c-Fos/activator protein (AP)-1 controls the expression of these cytokines by binding directly to AP-1 motifs in the cytokine promoter regions. T-5224 is a new drug developed by computer-aided drug design that selectively inhibits c-Fos/AP-1 binding to DNA. In this study, we tested whether T-5224 has a potential inhibitory effect against LPS-induced AKI, by suppressing the TNF-alpha inflammatory response and other downstream effectors.MethodsTo test this hypothesis, male C57BL/6 mice at 7 weeks old were divided into three groups (control, LPS and T-5224 groups). Mice in the control group received saline intraperitoneally and polyvinylpyrrolidone solution orally. Mice in the LPS group were injected intraperitoneally with a 6 mg/kg dose of LPS and were given polyvinylpyrrolidone solution immediately after LPS injection. In the T-5224 group, mice were administered T-5224 orally at a dose of 300 mg/kg immediately after LPS injection. Serum concentrations of TNF-alpha, interleukin (IL)-1beta, IL-6 and IL-10 were measured by ELISA. Moreover, the expression of intercellular adhesion molecule (ICAM)-1 mRNA in kidney was examined by quantitative real-time RT-PCR. Finally, we evaluated renal histological changes.ResultsLPS injection induced high serum levels of TNF-alpha, IL-1beta and IL-6. However, the administration of T-5224 inhibited the LPS-induced increase in these cytokine levels. The serum levels of IL-10 in the LPS group and T-5224 group were markedly elevated compared with the control group. T-5224 also inhibited LPS-induced ICAM-1 mRNA expression. Furthermore histological studies supported an anti-inflammatory role of T-5224.ConclusionsIn endotoxin-induced AKI, T-5224 inhibited the production of TNF-alpha and other downstream effectors. In contrast, T-5224 did not inhibit IL-10, an anti-inflammatory cytokine. These data support that the use of T-5224 is a promising new treatment for septic kidney injury.

A Phase I Trial of the Anti-Inhibitory KIR Antibody, IPH2101, and Lenalidomide in Multiple Myeloma: Interim Results

AbstractAbstract 4058 Introduction:Multiple myeloma (MM) cells upregulate MHC class I antigens which serve as ligands to inhibitory Killer Immunoglobulin-like Receptors (KIR) as a means of evading natural killer (NK) cell immunity. IPH2101 is a human, IgG4 monoclonal antibody against common inhibitory KIR that prevents KIR-ligand interaction and augments NK cell cytotoxicity against MM. A prior, single-agent, phase I study of IPH2101 demonstrated acceptable safety and tolerability with 34% of heavily pretreated patients achieving disease stabilization. Lenalidomide expands and activates NK cells, thus, pairing this agent with IPH2101 may represent a novel, steroid-free therapeutic option. Our preclinical data suggest that IPH2101 and lenalidomide combine to enhance NK cell function against MM. Herein, interim results of a phase I clinical trial of IPH2101 and lenalidomide are presented. Methods:A 3+3 design includes dose escalation over 4 planned cohorts: Cohort 1 = IPH2101 0.2mg/kg & lenalidomide 10mg, Cohort 2 = IPH2101 0.2mg/kg & lenalidomide 25mg, Cohort 3 = IPH2101 1mg/kg & lenalidomide 25mg, Cohort 4 = IPH2101 2mg/kg & lenalidomide 25mg. A 6-patient extension cohort is planned at the maximally tolerated dose. Lenalidomide is administered orally days 1–21 and IPH2101 is given intravenously on day 1 of a 28-day cycle for 4 cycles. Responding patients may continue therapy for 4 additional cycles and are maintained on lenalidomide alone thereafter. The primary objective of the study is to determine the safety and tolerability of IPH2101 and lenalidomide with secondary endpoints including: response rate, progression-free survival, pharmacodynamics, pharmacokinetics, and biologic correlates. Adult patients with relapsed MM following one or two lines of prior therapy are eligible to participate. Prior exposure to lenalidomide is allowed; however, patients must not have been resistant to, or intolerant of, lenalidomide. Patients must have measurable disease, ECOG performance status 0–2, and adequate organ function and bone marrow reserve. Thromboprophylaxis is mandatory for all patients. Results:To date, n=13 patients have been enrolled in the first 3 cohorts who have received a median number of 4 cycles of therapy to date (range 1–8). Adverse events have been generally low grade and self-limited. Two potential dose-limiting toxicities (DLT) have been observed (cohort 1 and cohort 3). Both patients experienced grade 3 infusion reactions associated with pyrexia and transient, grade 4 leucopenia following cycle 1, day 1 treatment. Clinical cytokine release syndrome was observed, and high levels of IFN-gamma, IL-6, TNF-alpha, and MIP-1beta were documented. Infusion reactions were treated with supportive care, including intravenous fluids and anti-pyretics; steroids were not administered. Leucopenia resolved without intervention, and both patients were subsequently treated in cycle 2 without recurrence of DLT. Both patients appear to have disproportionally greater KIR binding of IPH2101 on T cells relative to NK cells. Cohort 1 was expanded to n=6 patients without recurrence and expansion of cohort 3 is ongoing. An amendment is now included for pre-medication of patients with an anti-pyretic and anti-histamine prior to dosing. Co-administration of lenalidomide does not appear to alter the IPH2101 pharmacodynamic profile observed in the prior, single-agent phase I study. To date, responses include: 1 (unconfirmed) complete response, 3 partial responses, 2 minor responses and 1 stable disease. Conclusions:In addition to direct anti-MM effects, lenalidomide promotes NK cell activation, and IPH2101 prevents inhibitory signaling to augment NK cell immunity against MM. Results suggest the combination has activity with manageable toxicity in relapsed MM. At present, ongoing unconfirmed complete response has been > 12 months and ongoing partial responses have been > 18 months, > 8 months, and > 1 month in duration. This combination may represent the first steroid-free, “dual immunotherapy” for MM and further development is justified. Disclosures:Benson:Innate Pharma: Research Funding. Off Label Use: Lenalidomide without dexamethasone, IPH2101 not FDA approved yet. Zerbib:Innate Pharma: Employment. Andre:Innate Pharma: Employment.

166. Physical fitness decreases and central adiposity increases inflammation in normotensive and pre-hypertensive individuals

Elevated blood pressure (BP) has been associated with an increased inflammatory state; conversely, healthy lifestyle, leading to higher cardiovascular fitness and lower adiposity, is promoted to reduce inflammation. We examined relationships among cardiovascular fitness, adiposity, and circulating levels of TNF-alpha in 57 men and women with normal to mildly elevated (121/81–139/89 mmHg) BP. Cardiovascular fitness was determined by peak oxygen consumption (VO2peak) during a treadmill exercise test. Dual Energy X-ray Absorptiometry (DEXA) was used to determine percent total and abdominal fat. Plasma TNF-alpha levels were determined by commercial ELISA. Multivariate regression analyses were performed, predicting TNF-alpha with age, gender, systolic BP (SBP), diastolic BP (DBP), VO2peak, total percent fat and percent trunk fat in the model. Higher TNF-alpha levels were explained by lower VO2peak (beta = −.311; p = .042) and percent trunk fat (beta = .279; p = .023). These results reveal lower aerobic fitness and increasing percent trunk fat as significant predictors of elevated TNF-alpha levels among normotensive and pre-hypertensive individuals, beyond contributions of BP and total adiposity. Low-grade inflammation is strongly implicated in vascular pathology among individuals with elevated BP, although its exact mechanisms remain unclear. Our findings suggest an important role of adiposity, especially abdominal adiposity, in immune cell activation and constitutive cytokine production. Further research to elucidate the action of adiposity-derived factors (e.g, adipokines) in inflammation and immune activation is warranted.

Serum cytokine TNF-alpha and hemoglobin levels in Plasmodium falciparum malaria – A correlative study in coastal districts of Odisha

Background and objectivesThis study was conducted to assess the correlation between TNF alpha levels and Hemoglobin in patients affected by Plasmodium falciparum along with sex and age matched healthy controls. MethodsThis study included 32 patients of P. falciparum malaria, and 30 age and sex matched healthy controls. Blood samples were drawn from malaria patients admitted to medicine ward along with healthy controls and transferred to the Biochemistry department for the routine hematological parameters and ELISA for TNF-alpha, IL-12 and IL-4. ResultsThe levels of TNF-alpha were significantly elevated in falciparum malaria compared to healthy controls. Further Hb levels in Pf malaria showed significant decline compared to healthy controls. A strong negative correlation was observed between TNF-alpha and Hb levels in Pf malaria. ConclusionThe results indicate that in falciparum are characterized by up regulation of TNF-alpha and down regulation of Hb levels. Present study shows close association between severe anemia and high TNF-alpha levels.

P015 TSLP enhances type 1 immune response in cutaneous leishmaniasis patients

Introduction Leucocytes from cutaneous leishmaniasis (CL) patients due to L. braziliensis infection secrete high levels of IFN-gamma and TNF-alpha in response to soluble Leishmania antigen (SLA) and at lesion site. This type of immune response helps to control parasite multiplication but also mediates tissue damage and lesion development. Thus, we think CL patients would benefit from a balanced Th1/Th2 immune response. It has been shown that cells that do not classically participate in immune response, such as epithelial cells, can secrete soluble factors interfering on T cells responses to pathogens. Thymic stromal lymphopoietin (TSLP) is a recently described cytokine secreted by epithelial cells. TSLP-activated macrophages and dendritic cells enhance IL-4 and IL-13 secretion by memory CD4+ T cells in asthma and helminthes infections. Methods Peripheral blood mononuclear cells were obtained from CL individuals. Cells were stimulated with soluble Leishmania antigen or D. pteronyssinus in presence or absence of recombinant TSLP for 72 h. Cytokines were measured in cell culture supernatants by ELISA. For immunohistochemistry, biopsies were obtained from CL patients and stained for TSLP. Results Since TSLP induces Th2 responses, our hypothesis was that presence of TSLP would conditionate antigen presenting cells to enhance SLA-induced Th2 responses in CL patients decreasing levels of Th1 cytokines. Surprisingly, our data shows that in presence of TSLP, SLA-stimulated PBMC from CL individuals enhance IFN-gamma production, instead. No change in SLA-induced IL-5, TNF-alpha and IL-10 was observed. To test the effect of TSLP on antigens known to induce Th2 responses, we stimulated cells from the same patients with D. pteronyssinus (Derp) antigen in presence or absence of TSLP. As expected, presence of TSLP enhanced Derp-induced IL-5 production. The fact that TSLP can be expressed by skin epithelial cells, led us to investigate whether there was presence of TSLP in lesion of CL patients. Immunohistochemistry reveled TSLP expression in epithelial cells, endothelial cells and macrophages from CL individuals. We finally asked if Leishmania infection would interfere on TSLP receptor expression in monocytes. L. braziliensis-infected monocytes from healthy individuals up-regulated TSLP receptor expression. Conclusion Our results show a novel role for TSLP in inducing up-regulation of IFN-gamma production in CL patients. These data suggest that presence of TSLP may contribute for parasite killing and also maintenance of inflammatory response in these patients.

P030 Subsets of circulating monocytes differently contribute to immunopathology in cutaneous leishmaniasis

Introduction Cutaneous leishmaniasis (CL) caused by L. braziliensis is characterized by the presence of one or more ulcerated lesions with elevated borders. Cellular infiltrate in lesions of these individuals are composed mainly by mononuclear cells, including CD4+, CD8+, plasm cells and cells from myeloid lineage. High levels of IFN- γ and TNF- α are detected in these patients and these proinflammatory cytokines are known to play a role in the pathogenesis of CL, by inducing tissue damage. Upon infection with Leishmania or in presence of soluble Leishmania antigen, monocytes from CL individuals produce high levels of TNF-alpha and chemokines involved in recruitment of CD4+, CD8+ T cells and monocytes. Recent studies have shown that circulating monocytes constitute a heterogeneous population, and based on expression of CD14 and CD16, these cells can be divided in classical (CD14+CD16−), intermediate (CD14+CD16+) and non-classical (CD14−CD16+) monocytes. Intermediate and non-classical monocytes are known to migrate to inflamed sites and secrete inflammatory mediators, and high frequency of these cells has been associated with pathogenesis of many inflammatory diseases. TNF- α can mediate the pathology of the disease through various mechanisms including induction of nitric oxide, apoptosis, expression of metalloproteinases (MMPs) and increased cytotoxicity. MMP-9 is a zinc-dependent enzyme that degrades collagen type 4 (present in basal membrane) and has been associated with skin inflammatory diseases. Although the mechanism underlying ulcer development in CL is not known, it is likely that MMP-9 contribute to tissue damage since it was documented that L. braziliensis -infected macrophages upregulate MMP-9. Methods Peripheral blood mononuclear cells were obtained from patients with cutaneous leishmaniasis and uninfected controls. Cells were analysed ex-vivo or after culture in presence of Leishmania parasites, antigen or LPS, for MHC II, co-stimulatory molecules, TNF-alpha and MMP9. Results Thus, our goal was to investigate the contribution of sub-populations of monocytes to TNF-alpha and MMP-9 secretion in CL patients. We found that early after infection (pre-ulcerative phase) the frequency of intermediate and non-classical monocytes are elevated in blood of CL individuals. Also, while intermediate monocytes produced more TNF-alpha in response to Leishmania , non-classical ones were the main source of MMP-9 in most CL patients. Similarly, the biopsies studies reveled that the non-classical monocytes were the main MMP-9 producing cells. Conclusion These results show that monocytes subpopulations contribute differently to the immunopathology observed in CL patients.

High TNF-alpha and IL-8 levels predict low blood dendritic cell counts in primary cytomegalovirus infection

BackgroundIn vitro studies suggest that human cytomegalovirus (CMV) modulates the functions of dendritic cells (DCs). However, there are limited data on DC homeostasis in CMV-infected patients. ObjectivesThe aim of this study was to characterize circulating DCs and plasma cytokine levels in immunocompetent patients with primary, symptomatic CMV infections. Study designThe study population consisted of 14 patients suffering of CMV mononucleosis and 14 healthy volunteers (11 CMV-seropositive and 3 CMV-seronegative subjects) included as controls. Peripheral blood mononuclear cells were isolated and used to characterize DCs and to quantify CMV in the blood. Plasma levels of pro-inflammatory and anti-inflammatory cytokines were also measured. ResultsWe observed that patients who were developing CMV mononucleosis presented lower myeloid and plasmacytoid DC counts in peripheral blood compared with healthy controls. We also noted elevated levels of inflammatory mediators, of which tumor necrosis factor-α (TNF-α)—which activates DCs and endothelial cells—was the highest. Notably, the decrease in blood DCs correlated with high TNF-α and IL-8 levels by a hyperbolic function. ConclusionsOur results suggest that increased levels of inflammatory factors facilitate alterations in DC homeostasis during primary CMV infection, which may contribute to viral-induced modulation of host immunity.

TNF-alpha and IL-8: Serum levels and gene polymorphisms (−308G>A and −251A>T) are associated with classical biomarkers and medical history in children with sickle cell anemia

Sickle cell anemia (SCA) is a disorder characterized by a heterogeneous clinical outcome. In the present study, we investigated the associations between Tumor Necrosis Factor-alpha (TNF-alpha) −308G>A and Interleukin 8 (IL-8) −251A>T gene polymorphisms, medical history and classical biomarkers in children with steady-state SCA. In total, 210 SCA patients aged 2–21years and 200 healthy controls were studied. Gene polymorphisms, betaS-globin haplotypes and a 3.7-kb deletion in alpha2-thalassemia (α2-thal3.7kb) were investigated by PCR/RFLP analysis, and cytokine levels were determined by ELISA. Splenomegaly (p=.032) was more prevalent among children younger than 5years of age. The A allele of the TNF-alpha −308G>A gene polymorphism and the presence of α2-thal3.7kb were associated with an increase risk of splenic sequestration events (p=.001; p=.046), while the T allele of the IL-8 −251A>T gene polymorphism was considered to be a protective factor for splenomegaly events (p=.032). Moreover, the A allele of the TNF-alpha −308G>A gene polymorphism was associated with high TNF-alpha levels (p=.021), and the hemoglobin F and hemoglobin S haplotypes were correlated with serum levels of IL-8. The logistic regression analysis showed significant effects of the TNF-alpha and IL-8 gene polymorphisms, betaS-globin gene haplotypes and α2-thal3.7kb on the occurrence of splenic sequestration events. Our study emphasizes that the identification of new genetic and immunological biomarkers and their associations with classical markers is an important strategy to elucidate the underlying causes of different SCA phenotypes and their effects on patient outcome.

Modulation of intestinal morphology and immunity in nile tilapia (Oreochromis niloticus) by Lactobacillus rhamnosus GG

The use of lactic acid bacteria from human origins as a potential probiotic supplementation in aquaculture feed is now widely accepted. Here, we examined some of the properties and mechanisms of the action of Lactobacillus rhamnosus GG, originating from humans, on growth performance, gut mucosal immunity and humoral and cellular immune response in tilapia (Oreochromis niloticus). The results suggested that supplementation of L. rhamnosus gave an advantage in promoting the intestinal structure and the mucosal immunity of tilapia. Probiotic fish had a greater villous height in all parts of the intestines and, significantly, in the proximal and middle part. The population of intraepithelial lymphocytes was significantly higher in the probiotic group than in the control group in all parts of the intestines. The population of acidophilic granulocyte in the probiotic group was significantly higher at the proximal and distal parts when compared with the control group. The higher serum complement activity as well as the enhanced phagocytosis and killing ability of the head kidney leukocytes in the probiotic supplemented fish corresponded with the higher level of TNF alpha and IL-1 gene expression, suggesting that the induction of IL-1 and TNF alpha cytokines by L. rhamnosus served as an important regulator of gut associated immune systems.

P90 Injury frequence of target organs in preeclampsia

Background: Increased reactive oxygen species activity as well as high levels of tumor necrosis factor alpha (TNF-alpha) contribute to endothelial dysfunction and may be implicated in the pathogenesis of preeclampsia. Besides, hyperuricemia is a common finding in preeclamptic pregnancies. Aims: This study investigated uric acid levels in serum, TNF-alpha, superoxide anion (O2) and hydrogen peroxide (H2O2) produced by monocytes, as well as superoxide dismutase (SOD) and catalase (CAT) activities in erythrocytes from women with preeclampsia (PE) comparing with normotensive (NT) pregnant women. Methods: Serum and peripheral blood monocytes from 30 preeclamptic and 30 NT pregnant women were obtained in the last trimester of pregnancy for determination of uric acid levels, and endogenous O2 and H2O2 released by monocytes. TNF-alpha production by monocytes was determined by ELISA, and activities of SOD and CAT were evaluated in erythrocyte lysate obtained from both groups of pregnant women. Results: In preeclamptic group serum uric acid levels were significantly higher (6.1±1.5 vs 2.8±0.3 mg/dL) as well as endogenous O2 (2.17±0.18 vs 1.68±0.13 nM) and H2O2 (1.79±0.54 vs 1.4±0.25 nM) released from monocytes than in NT group. Higher levels of TNF-alpha were produced by monocytes from patients with PE (41.3±8.8 vs 21.1±9.9 pg/mL) compared with NT group. In addition, SOD activity in erythrocytes was significantly higher in PE group (5969.2±151 vs 4834.7±168 U/g Hb) than in NT group. No significant differences between groups were observed in relation to CAT activity. Conclusions: Increased serum uric acid levels were associated with O2 and H2O2 release and TNF-alpha production by monocytes in preeclampsia, and may contribute to enhance the oxidative and inflammatory state in this disorder. FAPESP: Grant no. 2006/07095-3

Comparison of Systemic Cytokine Levels in Mycobacterium spp. Seropositive and Seronegative Asian Elephants (Elephas maximus)

Mycobacterium spp. infection is an important health concern for Asian elephant (Elephas maximus) populations worldwide. The disease is of particular concern considering its potential to affect not only the individual animal but also herd and public health. Although elephant tuberculosis susceptibility is poorly understood, immune function alterations are central to disease pathogenesis in other species and probably affect outcome of mycobacterial infections in elephants. Measurement of immune mediator (cytokine) levels within blood samples can provide information regarding immune function that may elucidate disease susceptibility. For this study, mRNA levels of interleukin (IL)-2, IL-4, IL-10, and IL-12; interferon (IFN)-gamma; tumor necrosis factor (TNF)-alpha; and transforming growth factor (TGF)-beta were measured using elephant-specific, real-time reverse transcription-polymerase chain reaction (RT-PCR) assays in RNA-preserved whole blood samples from 106 Asian elephants, 15% of which were Mycobacterium tuberculosis complex seropositive. The Elephant TB STAT-PAK (Chembio Diagnostics, Inc., Medford, New York 11763, USA), a novel lateral flow antibody detection assay developed for specific use in elephants, was used to determine serologic status for the study. Seropositive animals had higher levels of TNF-alpha and lower levels of TGF-beta than seronegative animals; these differences between groups were statistically significant when levels were analyzed as categorical variables. Trends toward higher levels of IFN-gamma and IL-4 and slightly lower levels of IL-10 and IL-12 were noted in the seropositive group, although differences between groups were not statistically significant. Presence of other inflammatory conditions was found to be a significant confounding variable in the analysis of the relationship between tuberculosis status and TNF-alpha levels, necessitating its inclusion in statistical models. Age and sex were not found to significantly affect the relationship between tuberculosis status and any of the cytokines measured. Interleukin-2 levels were below the sensitivity of the real-time RT-PCR assay irrespective of tuberculosis status. These findings provide a foundation for future research into the immunopathogenesis of elephant tuberculosis.

Tumor necrosis factor-&amp;alpha; serum levels in healthy smokers and nonsmokers

Tobacco smoking is the most important risk factor for chronic obstructive pulmonary disease (COPD) development. Inhaled cigarette smoke can induce tumor necrosis factor-alpha (TNF-alpha) production by alveolar macrophages, which in turn may enhance the production of metalloproteinases (MMPs). MMPs have been involved in mediating airway inflammation and lung destruction. We aimed to measure the TNF-alpha serum levels in healthy heavy smokers and healthy nonsmokers to determine the dose-response relationship based on the cigarette smoke exposure. We included in our study 43 healthy heavy smokers and 19 healthy nonsmokers (the control group). The smokers group was classified as less than one pack, one pack, and more than one pack per day. A clinical and paraclinical evaluation was performed in both groups, without any evidence of infection or COPD. The serum levels of TNF-alpha were assessed by ELISA. The TNF-alpha serum levels were significantly higher for the group of smokers compared to the group of nonsmokers (P < 0.004). We also noticed an increased TNF-alpha concentration in the serum of smokers with more than one pack per day compared with those with less than one pack per day (P < 0.03). There was a positive correlation between the serum level of TNF-alpha and tobacco smoke exposure. The high levels of TNF-alpha in the serum of smokers suggest an imbalance between the proinflammatory and anti-inflammatory factors as a result of tobacco smoke exposure. The concentration of TNF-alpha is elevated in the serum of healthy heavy smokers in a cigarette dose-dependent manner. We speculate that the serum level of TNF-alpha might be a useful biomarker for the selection of heavy smokers with a high risk of developing smoke induced pulmonary diseases.

Periodontitis and arthritis interaction in mice involves a shared hyper-inflammatory genotype and functional immunological interferences

Periodontitis (PD) and rheumatoid arthritis (RA) have been found to be clinically associated and to share the chronic nature of the inflammatory reaction associated with bone resorption activity. However, the mechanisms underlying such association are unknown. Therefore, we examined the basis of Actinobacillus actinomycetemcomitans- and Porphyromonas gingivalis-induced PD and pristane-induced arthritis (PIA) interaction in mice. Higher severity PD in the genetically inflammation prone acute inflammatory reactivity maximum (AIRmax) mice strain was associated with higher levels of TNF-alpha, IL-1beta, IL-17, matrix metalloproteinase (MMP)-13, and RANKL, whereas PD/PIA co-induction resulted in even higher levels of IL-1beta, IFN-gamma, IL-17, RANKL, and MMP-13 levels. Conversely, PD/PIA co-induction in AIRmin strain did not alter the course of both pathologies. PIA/PD co-induction resulted in altered expression of T-cell subsets transcription factors expression, with T-bet and RORgamma levels being upregulated, whereas GATA-3 levels were unaltered. Interestingly, PIA induction resulted in alveolar bone loss, such response being highly dependent on the presence of commensal oral bacteria. No differences were found in PIA severity parameters by PD co-induction. Our results show that the interaction between experimental PD and arthritis in mice involves a shared hyper-inflammatory genotype and functional interferences in innate and adaptive immune responses.

Prospective study of hemostatic alterations in children with acute lymphoblastic leukemia

In a group of newly diagnosed acute lymphocytic leukemia (ALL) children we evaluated a number of hemostatic and inflammatory markers at diagnosis and at different time points during chemotherapy for the remission induction to identify alterations in the plasma levels of prothrombotic markers before and during the course of chemotherapy. The following plasma markers were evaluated: thrombin-antithrombin complex (TAT), D-Dimer, plasminogen activator inhibitor 1 (PAI-1), antithrombin, fibrinogen, von Willebrand factor (VWF) antigen and high molecular weight VWF (HMW-VWF) multimers, P-selectin, tumor necrosis factor alpha (TNF-alpha), and interleukin 6 (IL-6). Plasma samples were collected at the following time points: at T0 (baseline) and T1 (+24 days of therapy), T2 (+36 days therapy), and T3 (+64 days therapy). The results show that, at diagnosis, ALL children presented with laboratory signs of increased thrombin generation and fibrin formation (i.e. high TAT and D-dimer levels), fibrinolysis inhibition (i.e. high PAI-1 level), endothelial activation (i.e., high HMW-VWF and soluble P-selectin levels) and inflammation (i.e. high TNF-alpha and IL-6 levels). After starting induction therapy, the thrombin generation markers and inflammatory cytokines significantly decreased. To the opposite, PAI-1 and P-selectin significantly increased, suggesting an insult by chemotherapy on the vascular endothelium. These effects were more evident during steroid administration. Symptomatic venous thromboembolism (VTE) episodes developed in two cases during induction therapy, which did not allow the evaluation of the predictive value for VTE of laboratory markers.

Effect of Pre‐adipocyte and Differentiated Adipocyte Conditioned Media on Galectin‐3 Expression in Mouse Intestinal Epithelial Cell

We hypothesized that obesity cause inflammation which, in turn, promotes colon carcinogenesis. To test the hypothesis mouse intestinal IEC‐6 cells were incubated in the presence of preconditioned media collected from culturing LPS‐stimulated macrophage, followed by comparing protein expression profile with control cells. The proteomic analysis revealed 45 differentially regulated proteins in mouse intestinal IEC‐6 cells under the inflammation‐mimicking condition. Among others, galectin‐3, a beta‐galactoside‐binding protein, was up‐regulated. Galectin‐3 has been reported to play anti‐apoptotic role in tumor progression and metastasis, and its over‐expression has been observed in several types of cancer such as colon cancer. In order to determine relationship between obesity‐induced inflammation and colon carcinogenesis, conditioned media (CM) from LPS‐stimulated or non‐stimulated 3T3‐L1 pre‐adipocytes or differentiated adipocytes were utilized to induce inflammation in IEC‐6 cells. Nitric oxide (NO) level was determined using Griess method and galectin‐3 expression was analyzed by western blot. Results showed enhanced generation of NO and upregulation of galectin‐3 in both LPS‐stimulated pre‐adipocyte and adipocyte. The increased expression of galectin‐3 in IEC cells might be caused by high levels of TNF‐alpha and IL‐6 in CM.