Higher Serum Levels Of TNF-α Research Articles

Serum and gene expression profile of cytokines in first-episode psychosis.

An inflammatory syndrome has been previously reported in chronic schizophrenia. The aims of this study were to investigate: (1) serum levels and leukocyte gene expression of cytokines in patients with first-episode psychosis and controls; and (2) possible causes of abnormal cytokine levels in first-episode psychosis, testing their association with psychosocial stressors, current nicotine and cannabis use, and duration of antipsychotic treatment. We recruited 24 first-episode psychosis patients and 24 healthy controls matched for age, gender, ethnicity and body mass index. Serum interleukin(IL)-1α, IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, Tumour Necrosis Factor- α (TNF-α), Interferon- γ (IFN-γ), vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), and monocyte chemotactic protein-1 (MCP-1) were analysed in all subjects. Leukocyte gene expression analyses were conducted only for those cytokines that were different between-groups in the serum analyses. Patients had significantly higher serum levels of IL-1α (effect size d=0.6, p=0.03), IL-1β (d=0.4, p=0.01), IL-8 (d=0.6, p=0.01) and TNF-α (d=0.7, p=0.05) and a trend for higher IL-6 serum levels (d=0.3, p=0.09) when compared with controls. Leukocyte m-RNA levels of IL-1α (d=0.6, p=0.04), IL-6 (d=0.7, p=0.01) and TNF-α (d=1.6, p<0.001), but not IL-1β and IL-8, were also significantly higher in patients. A history of childhood trauma was associated with higher TNF-α serum levels (p=0.01), while more recent stressful life-events were associated with higher TNF-α mRNA levels in leukocytes (p=0.002). In conclusion, first-episode psychosis is characterised by a pro-inflammatory state supported, at least in part, by activation of leukocytes. Past and recent stressors contribute to this pro-inflammatory state.

Bacterial Superantigens Enhance the In Vitro Proinflammatory Response and In Vivo Lethality of the TLR2 Agonist Bacterial Lipoprotein

Bacterial superantigens are Gram-positive exotoxins that induce proinflammatory cytokine release in vitro, cause lethal shock in vivo, and can be detected in the bloodstream of critically ill patients. They also have a powerful priming effect on the TLR4 agonist LPS. The aim of this study was to investigate the relationship between superantigens and the TLR2 agonist bacterial lipoprotein (BLP). Priming of human monocytes or PBMCs with superantigens significantly enhanced proinflammatory cytokine TNF-α and IL-6 release in response to BLP stimulation. The priming effect of superantigens could be blocked by inhibiting p38 MAPK during the priming phase as opposed to NF-κB or ERK inhibition. This was consistent with higher expression of the phosphorylated p38 after superantigen priming and BLP or LPS stimulation. C57BL/6 mice with superantigen priming (10 μg/mouse) when challenged with BLP (600 μg/mouse) exhibited substantially higher mortality (100%) compared with mice without superantigen priming (zero). Mice given superantigen alone did not demonstrate any signs of illness. Mice challenged with both superantigen and BLP had significantly higher levels of serum TNF-α and IL-6 compared with those of mice challenged with either agent alone. Depletion of the monocyte/macrophage subpopulation significantly reduced the mortality rate from 100 to 20% in superantigen-primed, BLP-challenged C57BL/6 mice, with a 5- to 10-fold decrease in serum TNF-α and IL-6. Our results demonstrate that bacterial superantigens enhance the in vitro proinflammatory cytokine release and in vivo lethality of BLP. This novel finding may help to explain the massive proinflammatory cytokine release seen in superantigen-mediated septic shock.

Effects of magnetic induction hyperthermia and radiotherapy alone or combined on a murine 4T1 metastatic breast cancer model

Purpose: The purpose of this study was to explore the effects of MIH and radiotherapy alone or combined on metastatic breast cancer and the underlying mechanisms.Materials and methods: A murine 4T1 metastatic breast cancer model was established and randomly assigned into four treatment groups: C (control), R (radiotherapy), MIH, and MIH + R. Tumour volume, lung metastasis, the expression of Bax and MMP-9, T cell subsets, serum cytokine levels, and mouse survival were evaluated.Results: Group MIH + R showed significantly reduced tumour volume, lung metastasis, improved survival and increased Bax expression compared to group R or MIH (P < 0.05). MMP-9 expression in the primary tumour tissue was significantly increased in group R compared to the other groups (P < 0.05), which could be brought down by combined MIH treatment. Group MIH + R showed significantly higher CD4+ T cell percentage as well as CD4+/CD8+ cell ratio than group R (P < 0.05). Group MIH + R showed significantly higher serum levels of TNF-α, IFN-γ and IL-2 than group R (P < 0.05).Conclusions: MIH not only promotes the tumour-cell killing effect of radiotherapy through Bax-mediated cell death, but also improves cellular immunity in mice under radiotherapy and decreases the potential of radiotherapy to enhance MMP-9 expression, which leads to significant improvement in lung metastasis and overall survival of mice under combined treatment of MIH and R. This study is the first to have explored the effect of combined hyperthermia and radiotherapy on tumour metastasis and the underlying mechanisms. It provides insights into the application of MIH as an adjuvant to radiotherapy for metastatic breast cancer.

The effect of hysterectomy or levonorgestrel-releasing intrauterine system on cardiovascular disease risk factors in menorrhagia patients: A 10-year follow-up of a randomised trial

To compare, whether women with menorrhagia, treated with either hysterectomy or LNG-IUS, differ in their cardiovascular risk profile during 10-year follow-up. A total of 236 women were randomized to treatment by hysterectomy (n=117) or LNG-IUS (n=119). Their cardiovascular risk factors were analyzed at baseline, at 5 years, and at 10 years. As 55 originally randomized to the LNG-IUS group had hysterectomy during the follow-up, all analyzes were performed by actual treatment modality. Waist circumference, body-mass index (BMI), blood pressure, and the levels of blood lipids, serum high-sensitivity CRP (hsCRP) and tumor necrosis factor alpha (TNF-α) were measured, and the use of medication for hypertension, diabetes, hypercholesterolemia, and ischemic heart disease was analyzed. After 5 years, an increase in the use of diabetes medication during the follow-up was only detected in the hysterectomy group (from 1.7% to 6.7%, P=0.008 vs from 5.1% to 8.4%, P=0.08), as well as they had significantly higher serum levels of TNF-α (108.59 pg/ml vs 49.02 pg/ml, P=0.001) and hsCRP (1.55 μg/ml vs 0.78 μg/ml, P=0.038) at 5- and 10-years. There was no difference between the groups in the use of cardiovascular medication, neither was there difference in blood pressure, waist circumference, BMI, or concentrations of blood lipids. Hysterectomy seems to be associated with increased levels of serum inflammatory markers and increased diabetes medication, which in turn, may predispose individual to future cardiovascular events.

Relation of Serum TNF-α and TNF-α Genotype with Delayed Cerebral Ischemia and Outcome in Subarachnoid Hemorrhage

The pathogenesis of delayed cerebral ischemia (DCI) after subarachnoid hemorrhage (SAH) remains obscure. The authors assessed the relationship of tumor necrosis factor alpha (TNF-α) and TNF-α gene polymorphisms with occurrence of DCI and poor outcome at 3months. Serum levels of TNF-α were measured every other day until discharge in 67 patients and the mean serum levels per patient during days 0-12 were dichotomized at the median value of the whole group. TNF-α genotyping was available in 31 patients and related to serum TNF-α by means of one-way ANOVA analysis. The authors calculated hazard ratio's (HR) with corresponding 95% confidence intervals (CI) for the association with DCI by means of Cox proportional hazard analysis and odds ratio's (OR) for the association with poor clinical outcome by means of logistic regression analysis. In both analyses the authors adjusted for sex, age, amount of blood, and clinical condition at admission. Leukocytes and CRP were investigated similarly for comparison. For high-serum TNF-α levels during days 0-12 adjusted HR was 0.6 (95%CI: 0.1-2.4) for DCI and adjusted OR 2.0 (95%CI: 0.4-9.0) for clinical outcome. Serum TNF-α levels were 11.4pg/ml for wildtype TNF-α genotype and 9.7pg/ml for the non-wildtype TNF-α genotype (P=0.15). For the non-wildtype TNF-α genotype the HR for DCI was 0.4 (95%CI: 0.1-2.6) and the OR for clinical outcome was 0.8 (95%CI: 0.1-4.0). It is unlikely that serum TNF-α or TNF-α genotype play an important role in the occurrence of DCI after SAH.

Increased lethal cytokine-mediated toxicity following immunotherapy in aged versus young mice (156.24)

Abstract We and others have previously demonstrated that agonistic αCD40 and IL2 immunotherapy (IT) treatment in young mice results in synergistic anti-tumor effects in multiple tumor models. However, efficacy of this and other ITs can be limited by their induction of systemic toxicity. The majority of mouse tumor studies use younger mice (8-12 weeks) which do not accurately represent the age of a typical cancer patient (&amp;gt;50 yrs or 12 months in a mouse). Therefore, we sought to determine the potential toxicity of IT in aged versus young mice. Within 2-3 days after IT treatment, marked toxicity was observed in aged (17-22 months) and middle aged mice (9-12 months) resulting in 100% mortality as opposed to no mortality in young mice (2-4 months). Aged mice had significantly higher serum levels of TNFα, IFNγ, IL6 and IL17a after IT compared to young treated mice, with a proportionate increase with age. Histological evaluation of aged treated mice revealed disseminated lymphocytic infiltrates to the gut, lung and liver; multiple organ damage with hepatic piecemeal necrosis; extensive GI mucosal damage and peribronchiolitis in the lung compared to young treated mice. Finally, IT-treated aged IFNγ-receptor and TNFα-KO mice showed significant decreases in mortality compared to treated WT-aged mice. This data suggests that IT with αCD40/IL2 induces a cytokine storm culminating in multiple organ damage and rapid lethality that correlates with increased serum cytokines and worsens with age.

Aging Linked to Type 2 Diabetes Increases Oxidative Stress and Chronic Inflammation

Oxidative stress (OxS) and inflammation are physiopathological mechanisms related to diabetes and aging. We evaluated the additive effect of diabetes and aging on OxS and inflammation in a cross-sectional comparative study of 228 subjects: (1) 56 healthy adults (mean age, 47 ± 7 years); (2) 60 diabetic adults (mean age, 52 ± 6 years); (3) 40 healthy elderly adults (mean age, 67 ± 7 years); and (4) 72 diabetic elderly adults (mean age, 68 ± 7 years). We measured levels of glycosylated hemoglobin (HbA1c), plasma lipid peroxides, superoxide dismutase, glutathione peroxidase, total antioxidants, and tumor necrosis factor-alpha (TNF-α). The results indicate that diabetes is a risk factor for subjects with high serum levels of TNF-α (odds ratio [OR] = 12.1; 95% confidence interval [95% CI], 5.0-28; p < 0.001); this correlation becomes stronger when it is also associated with aging (OR = 14; 95% CI, 3.7-53.7; p < 0.05). Likewise, we observed that diabetes is an independent risk factor for OxS (OR = 2.1; 95% CI, 1.2-3.8; p < 0.05), and a stronger factor in older patients (OR = 3.1; 95% CI, 1.3-7.5; p < 0.05). Our findings suggest that aging, in concert with diabetes, exerts an additive effect on OxS and inflammation.

Characteristic Proinflammatory Serum Cytokine Profiles In Patients with B-Cell Chronic Lymphocytic Leukemia

Abstract Abstract 3595 Background: Cytokines are posited to play a critical regulatory role on the survival of the B-cell neoplastic clone in Chronic Lymphocytic Leukemia (CLL). AIM: The primary goals of this study were 1) to define additional relevant cytokines, growth factors and chemokines in CLL pathophysiology and 2) to correlate abnormal cytokine levels with disease stage, relevant hematological data and multiple prognostic factors. METHODS: A novel bead-based protein array system was employed to simultaneously measure 38 proinflammatory cytokines in the sera of CLL patients (N=116) and healthy age and sex matched controls (N=30). These results were correlated with Rai stage, β2-microglobulin level, LDH, CD38 expression and cytogenetic abnormalities. RESULTS: We first confirmed previous observations that TNFα, IL-1α, IL-1RA, IL-10, sIL-2Rα, VEGF and sCD40 ligand are significantly elevated in patients with CLL as compared with healthy controls. Expanding on the current literature, we demonstrated perturbations in an additional 15 serum cytokines in affected individuals. Compared to healthy controls, CLL patients had an increase in serum levels of IL-3 (p=0.002), IL-7 (p=0.008), INF-2α (p&lt;0.0001), MCP-1 (p&lt;0.0001), MIP-1β (p=0.002), MDC (p&lt;0.0001), Fractakine (p&lt;0.0001), EGF (p&lt;0.0001), FGF-2 (p&lt;0.0001), GRO (p&lt;0.0001), Eotaxin (p&lt;0.0001) and FLT-3 ligand (p&lt;0.0001). Patients with CLL also exhibited significantly lower levels of INF-γ (p&lt;0.01), IL-6 (p&lt;0.005) and IL-8 (p&lt;0.002) when compared to healthy individuals. Advanced Rai stage and high risk chromosomal abnormalities (del 11q and del 17p) strongly correlated with higher serum levels of TNFα, soluble IL-2Rα, IL-10, MCP-1, MIP-1α, MIP-1β and IP-10. Finally, serum levels of TNFα, MIP-1α and MIP-1β correlated with other adverse prognostic markers, including total white blood cell count, serum β2-microglobulin and LDH levels as well as CD38 expression. CONCLUSION: We have demonstrated numerous previously unrecognized cytokine abnormalities in patients with CLL and described a unique cytokine signature associated with advanced disease. Supported by the current understanding of cytokine biology and CLL pathophysiology, our observations suggest an important regulatory role for hematopoietic cytokines, such as IL-3 and IL-7, in promulgating survival of the aberrant B-cell clone. Likewise, the profoundly high levels of chemokines (i.e. MCP-1, MIP-1α, MIP-1β, IP-10) and their association with high risk prognostic factors argue for their role in sustaining the neoplastic microenvironment. Finally, the altered levels of IL-10, IL-6 and INF-γ observed in patients with CLL likely contribute to the immunosuppressive phenotype of the disease state. Disclosures: No relevant conflicts of interest to declare.

The TNF −308G>A Polymorphism Predicts Outcome of Patients with B-Cell Chronic Lymphocytic Leukemia In Relation to the IgVH Mutation Status

Abstract Abstract 2421 Background: Tumor necrosis factor (TNF)–α is an important pro-inflammatory cytokine involved in the modulation of lymphoma development and the balance between cell-mediated and humoral immunity. Deregulated concentrations of TNF–α have been detected in patients with lymphoma and were associated with an adverse prognosis. Evidence that the single-nucleotide polymorphism (SNP) in TNF promoter, TNF −308G&gt;A, could be the susceptibility locus for non-Hodgkin lymphoma (NHL) has been provided by case-control studies. Therefore we tested the hypothesis that TNF −308G&gt;A influences clinical course of B-cell chronic lymphocytic leukemia (CLL). Patients and Methods: We genotyped TNF −308G&gt;A (rs1800629) in 278 newly diagnosed patients with CLL and 192 ethnically-matched healthy individuals using the 7900 HT Real-Time (Applied Biosystems, USA). Some randomly selected DNA samples were analysed by direct sequencing using 3130xl Genetic Analyzer (Applied Biosystems, USA). Sequence data were based on the NCI SNP500 website. The IgVH mutation status in CLL patients was performed according to the protocol described by van Dongen et al. Serum samples from the 153 newly diagnosed CLL patients were collected at the time of diagnosis and tested by an enzyme-linked immunosorbent assay (ELISA) kit for human TNF (Quantikine, R&amp;D Systems, USA). Results: The TNF −308G&gt;A allelic frequencies and distributions were consistent with Hardy-Weinberg equilibrium, and did not differ significantly between CLL patients and the control group. There were no significant differences between TNF allelic or genotype distributions and clinical characteristics of CLL patients at diagnosis, including age, clinical stage according to Rai classification, serum LDH and β2-microglobulin levels, surface CD38 expression, ZAP-70 expression, Döhner's cytogenetic groups, and IgVH mutation status. Neither of assessed TNF polymorphic variants was associated with response to first-line treatment, progression free survival (PFS) nor treatment free survival (TFS) that was measured from time point of diagnosis to first therapy. With a median follow-up of surviving patients of 52 months (range 1–209 months), the group of patients with TNF (−308A) allele (TNF−308AA or TNF−308AG genotypes) had significantly shorter overall survival (OS) compared to those carrying TNF (−308GG) genotype (p=0.01, log–rank test). To further characterize the prognostic impact of genetic variation in TNF on CLL patients survival, we divided the patients according to the IgVH mutation status into a IgVH unmutated (homology ≥98%) and a IgVH mutated (homology &lt;98%) group. We found that the patients carrying TNF (−308A) allele presented significantly shorter OS in the IgVH unmutated group compared to those with TNF (−308GG) genotype (p=0.02). Of note, the TNF −308G&gt;A polymorphism did not influence survival of patients with the IgVH mutated gene. To further investigate this difference, we analyzed the impact of TNF SNP on serum TNF-α levels in CLL patients at the time of diagnosis. We found that high TNF-α levels, greater than the median (16.84 pg/mL) value, correlated with stages III and IV according to Rai classification (p=0.001, chi2 test), elevated serum levels of LDH (p=0.01) or β2-microglobulin (p=0.001), CD38 expression ≥30% (p=0.001), ZAP-70 expression &gt;20% (p=0.01) as well as with TNF (−308AA) genotypes (p=0.04). The patients with high TNF-α levels had significantly shorter TFS (p&lt;0.0001) compared to those with low cytokine levels. This association retained its prognostic impact on TFS both in the IgVH unmutated (p=0.004) and mutated (p&lt;0.0001) group. No correlations between serum TNF-α levels and response to first-line treatment or PFS were found. Furthermore, in the group with high TNF-α levels, OS was significantly shorter compared to those in the cytokine low level group (p=0.04). Interestingly, when the patients were stratified according the IgVH mutation status, the high serum TNF-α level retained its prognostic impact on shorter OS only in the IgVH unmutated group compared to the patients with low TNF-α levels (p=0.04). Conclusions: Our results indicate that the TNF −308G&gt;A polymorphism along with the serum TNF-α level may influence CLL outcome especially in the IgVH unmutated subgroup, which points out the importance of innate immunity genes for CLL variability and prognosis. Disclosures: No relevant conflicts of interest to declare.

Children with familial hypercholesterolemia are characterized by an inflammatory imbalance between the tumor necrosis factor α system and interleukin-10

Familial hypercholesterolemia (FH) is associated with increased risk of premature atherosclerosis. Increasing evidence supports involvement of inflammation in atherogenesis. The inflammatory cytokine tumor necrosis factor (TNF)α has been regarded as a key mediator in the development of atherosclerosis due to its involvement in several stages in this process. We hypothesized that children with FH, as a model of early atherosclerosis, have different serum levels of inflammation markers than healthy control children. We measured serum levels of TNFα, as well as its endogenous inhibitors (i.e., soluble TNF receptors [sTNFR] 1 and 2) and the anti-inflammatory cytokine interleukin (IL)-10 in healthy children (7-20 years) with (n=102) and without (n=48) heterozygote FH as well as adult FH subjects (n=20) and healthy adult controls (n=16). The main findings were: Compared to control children, FH children had higher serum levels of TNFα, accompanied by lower sTNFRs levels, resulting in an increased TNFα/sTNFRs ratio (P<0.05), potentially reflecting enhanced TNFα activity. In contrast to the increased TNFα levels, FH children had decreased serum levels of IL-10 (P<0.01) resulting in an increased TNFα/IL-10 ratio (P<0.01). We did not observe any difference in the same parameters between adult subjects with and without FH. FH children are characterized by an inflammatory imbalance between TNFα and IL-10, potentially contributing to the accelerated atherosclerotic process in these individuals.

High serum levels of TNF-α and IL-6 predict the clinical outcome of treatment with human recombinant erythropoietin in anaemic cancer patients

A number of anaemic cancer patients are not responsive to treatment with recombinant human erythropoietin (rHuEPO). The aim of the present study is to investigate whether serum levels of tumour necrosis factor-alpha (TNF-alpha), interleukin (IL)-1beta, IL-6 and additional laboratory parameters, together with clinical variables, can predict the clinical outcome of treatment with rHuEPO in anaemic cancer patients. Thirty-five cancer patients and 25 healthy controls were enrolled in this study. Patients were treated with epoetin alfa at the dose of 150 IU/kg s.c. three times a week for 12 weeks. If the haemoglobin (Hb) level failed to improve at least 2 g/dl above baseline by week 6 of treatment, dose was increased to 300 IU/kg s.c. for the remainder of the treatment period. All patients filled out the Brief Fatigue Inventory (BFI), a questionnaire for the self-evaluation of cancer-related fatigue. Serum samples from patients and control groups were frozen at -80 degrees C and TNF-alpha, IL-1beta and IL-6 were later examined by enzyme-linked immunosorbent assay. Fatigued cancer patients had significant higher levels of circulating TNF-alpha, IL-1beta and IL-6 than healthy controls. Responders (Rs) to erythropoietin had significant lower medium levels of TNF-alpha and IL-6 than nonresponders (NRs). Fatigued patients with a general BFI score > or =6 presented higher medium level of cytokines than nonfatigued patients (general BFI score <6), but each group responded similarly to treatment with rHuEPO. High serum levels of TNF-alpha and IL-6 at the baseline are significantly correlated with a negative response to administration with rHuEPO. Thus, pretreatment evaluation of TNF-alpha and IL-6 serum levels can help to select those patients who are most likely to benefit from treatment with rHuEPO. On the contrary, Hb level, red blood cell count, lactate dehydrogenase and BFI score do not predict the outcome of treatment with rHuEPO.

Association of Helicobacter pylori infection with systemic inflammation in preeclampsia

Objective. The aim of this study was to compare C-reactive protein (CRP), tumor necrosis factor alpha (TNFα), Chlamydia pneumonia IgG, IgM and plasma Helicobacter pylori IgA levels between preeclamptic and normal pregnant women and to determine whether seropositivity to Helicobacter pylori is associated with elevated levels of CRP and TNF-α.Methods. Forty patients with preeclampsia and 40 normotensive pregnant women of similar age and body mass index at the third trimester of gestation were selected for the study. Chlamydia pneumonia IgM and IgGs, Helicobacter pylori IgAs and concentrations of CRP and TNF-α were measured.Results. Concentrations of CRP and TNF-α were significantly higher in patients with preeclampsia than in control subjects. In the preeclamptic group, positivity rate for Helicobacter pylori IgA was significantly higher as compared to controls (p = 0.034). CRP and TNF-α levels were higher in Helicobacter pylori seropositive subjects.Conclusion. We demonstrated high levels of serum CRP and TNF-α in preeclamptic women who were seropositive to Helicobacter pylori in comparison with those in seronegative subjects.

Association of Obesity and Breast Cancer Risk: The Role of Estrogen, Tumor Necrosis Factor-alpha, and Adiponectin as Risk factors (preliminary study)

BACKGROUND: Breast cancer is the most frequent cancer diagnosed among women. Many factors influence the carcinogenesis of breast cancer. The aim of this study to analyze the role of obesity (waist circumference and body mass index), serum Estradiol levels, TNF-α, and Adiponectin in the occurrence of breast cancer.METHODS: This was observational study with casecontrol design. Eleven breast cancer patients as cases and twelve Fibroadenoma Mammae (FAM) patients as controls were analyzed. The serum Estrogen, TNF-α and Adiponectin were examined in their association with breast cancer risk.RESULTS: Women with breast tumor and waist circumference &gt; 80 cm have significantly higher breast cancer risk than women with breast tumor and waist circumference &lt;80 cm (OR 8.75; 95% CI=1.24-61.88; p=0.029). Women with breast tumor and higher serum TNF-α levels (&gt;2.30 pg/ml) have higher breast cancer risk (19.25 times) than women with breast tumor and have lower serum TNF-α levels (95% CI=1.77-209.55, p=0.015). Whereas, women with breast tumor and lower Adiponectin/TNF-α ratio (&lt; 2.13) have higher breast cancer risk (22.5 times) than women with breast tumor and higher Adiponectin/TNF-α (95% CI=2.60-194.51; p=0.005).CONCLUSION: These results suggest that high concentration of serum TNF-α, waist circumference &gt;80 cm and low Adiponectin/TNF-α ratio in women with breast tumor are significantly associated with an increased risk for breast cancer.KEYWORDS: Obesity, breast cancer, adiponectin/TNF-α ratio

Mycobacterium avium infection in HIV-1-infected subjects increases monokine secretion and is associated with enhanced viral load and diminished immune response to viral antigens.

The complex interaction between HIV-1 infection and Mycobacterium avium was studied. Viral burden was assessed, as well as immune response to HIV-1 in the context of Myco. avium infections. We also examined serum cytokine levels and cytokine release by blood mononuclear cells in HIV-1-infected subjects, infected or not with Myco. avium. Undetectable serum levels of IL-1, tumour necrosis factor-alpha (TNF-alpha) and IL-6 were found in normal controls and in groups I, II and III of HIV-1-infected subjects. Moderate levels of TNF-alpha, IL-1 and IL-6 were found in the sera of group IV patients. When group IV was subdivided into subjects with and without Myco. avium infections, subjects with Myco, avium infections were shown to have higher serum levels of TNF-alpha, IL-1 beta and IL-6 than those with other infections. Blood mononuclear cells from controls and HIV subjects were stimulated with bacterial lipopolysaccharide, and cytokine levels assessed. Cells from group II patients were shown to secrete normal levels of TNF-alpha and IL-6, and lower levels of IL-1 beta; group III subjects released higher levels of IL-6. Patients in group IV had blood cells that released elevated levels of IL-6 and TNF-alpha, and lower levels of IL-1 beta. Group IV subjects with Myco. avium infections had blood cells that released higher levels of TNF-alpha, IL-6 and IL-1 than group IV subjects with other infections. Assessment of viral burden in cells of HIV-1-infected subjects revealed that Myco. avium-infected subjects had a higher level of virus burden and a lower level of lymphoproliferative response to an inactivated gp120-depleted HIV-1 antigen than AIDS subjects with other infections. These data suggest that Myco. avium infections in HIV-1-infected subjects hasten the progression of viral disease, enhance cytokine release and contribute to the anergy to viral antigens.

Relationship of serum TWEAK level to cytokine level, disease activity, and response to anti‐TNF treatment in patients with rheumatoid arthritis

Objectives: To determine the serum concentration of tumour necrosis factor (TNF)‐related weak inducer of apoptosis (TWEAK) in patients with rheumatoid arthritis (RA) and to investigate the relationship between TWEAK level and disease activity, proinflammatory cytokine levels, and response to anti‐TNF treatment.Methods: Serum samples from 40 patients with RA, 40 patients with ankylosing spondylitis (AS), and 40 healthy subjects were collected. Serum samples from 26 patients with RA who received etanercept treatment were also collected in the 12th week of etanercept therapy. Serum TWEAK, TNFα, and interleukin (IL)‐6 levels were determined by enzyme‐linked immunosorbent assay (ELISA), and disease activity of RA was assessed according to the 28‐joint count Disease Activity Score (DAS28).Results: Patients with RA had significantly higher serum levels of TWEAK, TNFα, and IL‐6 compared with controls (p<0.05). Patients with AS also had significantly higher serum levels of TNFα and IL‐6 (p<0.05), but their serum TWEAK levels were not different from those of the controls. In patients with RA, serum TWEAK levels correlated with DAS28 (r2 = 0.452, p = 0.012) and TNFα levels (r2 = 0.653, p<0.001) but not with IL‐6 levels. Among RA patients who were treated with etanercept, responders showed a significant decrease in serum TWEAK levels at the 12th week of treatment, whereas TWEAK levels in nonresponders were not different from their baseline levels.Conclusions: Serum levels of TWEAK were significantly elevated in patients with RA, and reflected disease activity and short‐term response to etanercept treatment.

Tumor Necrosis Factor-α and Vascular Angiotensin II in Estrogen-Deficient Rats

Alterations in the vascular angiotensin II system may play a role in the pathophysiology of vascular disease after menopause. In previous studies we have shown that an increase in tumor necrosis factor (TNF)-alpha levels in aging rats because of estrogen deficiency may result in vascular dysfunction. In this study we investigated the effect of TNF-alpha inhibition in angiotensin II modulation of vascular function in aging female animals. Female rats approaching reproductive senescence (12 to 15 months old) were ovariectomized and treated with placebo, estrogen, or a selective TNF-alpha inhibitor (etanercept) for 4 weeks. Expression of angiotensin II in mesenteric arteries was evaluated by immunofluorescence, and the expression of angiotensin-converting enzyme and angiotensin type I receptor (AT(1)R) was investigated by Western immunoblot. Vascular function was assessed in mesenteric arteries using the myograph system, and the role of endogenous angiotensin II on adrenergic vasoconstriction was evaluated in vitro by selective AT(1)R blockade (Candesartan; 10 micromol/L). Our data demonstrate that estrogen-depleted rats have higher serum levels of TNF-alpha and greater sensitivity to phenylephrine vasoconstriction compared with estrogen-replaced animals, which was attenuated by AT(1)R blockade. In vivo TNF-alpha inhibition or estrogen replacement reduced phenylephrine constriction of mesenteric arteries and decreased the modulation of this vasoconstriction by candesartan. These functional changes were accompanied by a reduction in the vascular expression of angiotensin II, angiotensin-converting enzyme, and AT(1)R. These observations indicate that upregulation of TNF-alpha during estrogen deficiency may contribute to enhance vascular constriction by altering the vascular angiotensin II system.

Increased pathology in lungs of mice after infection with an α-crystallin mutant of Mycobacterium tuberculosis: changes in cathepsin proteases and certain cytokines

Latency and reactivation are a significant problem that contributes to the incidence, transmission and pathogenesis of tuberculosis. The mechanisms involved in these processes, at the level of both the bacillus and the host, are poorly understood. In Mycobacterium tuberculosis the alpha-crystallin (acr) gene has been linked to latency, because it is highly expressed during hypoxic growth conditions. Deletion of the acr gene in M. tuberculosis H37Rv (Deltaacr strain) was previously shown to reduce the intracellular growth of bacilli in macrophages; however, its impact on pathogenesis in vivo was unknown. This study demonstrated that infection of C57BL6 mice with Deltaacr results in lung bacillary loads 1-2 log units higher in comparison to parental H37Rv. Haematoxylin/eosin staining of lungs revealed exacerbated pathology characterized by extensive obliteration of alveolar air spaces by granulomatous inflammation. RT-PCR analysis and immunostaining of lungs showed that infection with either H37Rv or Deltaacr results in the differential expression of lysosomal cathepsin proteases. A slight increase in the expression of the matrix-degrading acidic-type cathepsins B, D and H was noted in Deltaacr-infected mice and was associated with clusters of macrophages within lung granulomas. Deltaacr-infected mice also showed high serum levels of TNF-alpha, IFN-gamma and G-CSF, suggesting that Acr may play a role in modulating the host response to infection.

Evaluation of tumour necrosis factor-α, soluble P-selectin, γ-glutamyl transferase, glutathione S-transferase-π and α-fetoprotein in patients with hepatocellular carcinoma before and during chemotherapy

Hepatocellular carcinoma (HCC) is an environmentally related cancer, with both viral and chemical carcinogens involved in a multistage process. To date, it has been difficult to detect the asymptomatic precursor lesions in early HCC. Therefore, the majority of HCC patients are not amenable to therapy, as they are detected at late stages. To evaluate the significance of tumour necrosis factor-α (TNF-α), sP-selectin, γ-glutamyl transferase (GGT), glutathione S-transferase-π (GST) and α-fetoprotein (AFP) in the diagnosis and follow up of HCC patients during chemotherapy with adriamycin, 45 subjects (15 healthy volunteers, 15 with benign liver diseases and 15 HCC patients) are studied before and during chemotherapy (three cycles of intravenous adriamycin). HCC patients had significantly higher serum levels of TNF-α, sP-selectin, GGT, GST and AFP. Serum levels of GGT and GST were significantly higher in HCC patients with poorly differentiated tumours than in patients with well- and moderately differentiated tumours. Treatment with adriamycin for three cycles produced a significant decrease in TNF-α, sP-selectin and GST. Thus, it is concluded that GST is a superior diagnostic indicator and may be a prognostic marker in HCC patients.

High serum TNF-α level in Type 2 diabetic patients with microangiopathy is associated with eNOS down-regulation and apoptosis in endothelial cells

A high dose of tumor necrosis factor (TNF)-alpha induces endothelial dysfunction and enhances apoptosis in vitro. The present study was conducted to examine whether incubating human umbilical vein endothelial cells (HUVECs) with serum from Type 2 diabetic patients complicated with retinopathy and/or microalbuminemia demonstrate endothelial dysfunction. Serum levels of TNF-alpha and vascular endothelial growth factor (VEGF) were elevated in diabetic patients. Plasma levels of TNF-alpha, two soluble TNF-alpha receptors (sTNFR), and VEGF were assessed in diabetic patients (CD, n=21) complicated with retinopathy and/or nephropathy, uncomplicated diabetic patients (UD, n=18), and in healthy normal participants (NS, n=16). In HUVECs incubated with patient's serum, endothelial constitutive nitric oxide synthase (eNOS) protein expressions were measured by Western blot analysis. Apoptosis in HUVECs was determined by optical microscopy, DNA fragmentation, and CPP32-like protease activity. Serum TNF-alpha, sTNFR-I, and asymmetric dimethylarginine (ADMA), an endogenous inhibitor of NOS, in CD were significantly higher than in UD or NS. While, serum sTNFR-I and VEGF levels were significantly increased in the both diabetic patients, compared with those of NS, no difference was observed in the serum TNF-alpha, sTNFR-II, and ADMA levels between UD and NS. eNOS down-regulation and apoptosis were seen in HUVECs incubated with serum from CD for 24 h, but those observations were completely counteracted in the incubation by the addition of the antihuman TNF-alpha antibody. These results imply that eNOS down-regulation in CD is associated with high serum TNF-alpha levels despite of high serum of VEGF levels. Therefore, endothelial dysfunction in diabetic patients complicated with microangiopathy may, in part, be attributed to high serum TNF-alpha levels.

Antitumour effect of Lactobacillus casei CRL 431 on different experimental tumours

We analysed the mechanisms of tumour inhibition by Lactobacillus casei CRL 431 orally administered, for fibrosarcome (non intestinal tumour, NIT) and for carcinoma (intestinal tumour, IT), both induced chemically. Balb/c mice were fed with L. casei (1.2×109cel/day/mouse) for two consecutive days previous to the induction of the respective tumours. After IT induction, oral administration of L. casei was repeated cyclically every 5 days for 5 months. In order to determine the percentage of tumour inhibition in the NIT mice, morphometric studies were performed and TNFα levels in serum and spleen cells were determined. In the IT, macroscopic and histopathological studies were performed and the number of intestinal IgA+ and TNFα+ cells was determined. We demonstrated that L. casei CRL 431 inhibited the growth of both tumours. In the case of NIT animals treated with L. casei, the high levels of serum TNFα would play an important role in the inhibition of tumour growth due to its cytolytic activity against tumour cells. In the IT, the increase in IgA+ and TNFα+ cells could be involved in the inhibitory effect observed. Our results showed that the selection of the probiotic bacteria with antitumour activity would not be limited to the determination of the cytokines relevant to this activity because of the multifactorial etiology of the different cancers.