Curcumin reduces the expression of interleukin 1β and the production of interleukin 6 and tumor necrosis factor alpha by M1 macrophages from patients with Behcet's disease

Objective: Behcet's disease (BD) is an auto-inflammatory disorder. Curcumin as a bio-active agent has anti-inflammatory properties. Effects of curcumin on the pathogenesis of BD are still not clear. In this study, we investigated the effect of curcumin on the inflammatory cytokines expression and production in M1 macrophages from BD patients compared with healthy controls.

Methods: Monocytes were collected from 10 healthy controls and 20 active BD patients, differentiated to macrophages by macrophage-colony stimulating factor for 7?d. Macrophages were then treated with interferon gamma, lipopolysaccharide, and curcumin (10 or 30?µg/ml) for 24?h. Analysis of tumor necrosis factor-alpha (TNFα), interleukin 1β (IL-1β), and IL-6 mRNA expression and protein production was performed using SYBR Green qPCR and ELISA method.

Results: Treatment with 30?µg/ml curcumin significantly down-regulated mRNA expression of IL-1β (p?<?.05) and protein production of IL-6 (p?p?p?Conclusions: We demonstrated that curcumin can inhibit the expression and production of inflammatory cytokines in M1 macrophages from BD patients. Our results suggest that curcumin can modulate inflammatory signaling more specifically in macrophages from BD patients than healthy macrophages.     

Recombinant human interleukin 1β and tumor necrosis factor affect glycosylation of serumα 1-acid glycoprotein in rats

Serum concentration and glycosylation of rat ₁,-acid glycoprotein ( ₁-AGP) were evaluated after the in vivo administration of recombinant human interleukin-1 (rhIL-1) and tumor necrosis factor (rhTNF-), alone or associated. The effect of LPS and turpentine was also studied. In all models, serum ₁-AGP concentrations were increased and glycosylation was altered. The ₁-AGP levels reached 1.8 g/liter with cytokines alone, 2.1 g/liter with cytokines associated or LPS, and 3.4 g/liter with turpentine. Analysis by concanavalin A (Con A) affinoimmunoelectrophoresis (CAIE) revealed that the relative proportion of Con A unreactive form always decreased whatever the inducing agent. On the other hand, the resulting effect on the concentrations cf Con A unreactive ₁-AGP concentrations was an increase with cytokines alone or LPS and a decrease with cytokines associated or turpentine. These results suggest a dissociation between the alteration in the level of ₁-AGP synthesis and in the pattern of its glycosylation in the various inflammatory models.     

Provider recommendations and maternal practices when providing breast milk to children with immunoglobulin E–mediated food allergy

BackgroundThere is limited research investigating maternal dietary practices and health care provider recommendations when providing breast milk (BM) to children with immunoglobulin (Ig) E–mediated food allergy.ObjectiveTo explore health care provider recommendations and maternal practices when providing BM to children with IgE-mediated food allergy and to assess for possible IgE-mediated reactions to BM while the mother consumed the food to which her child has allergy.MethodsA web-based survey was distributed to breastfeeding (BF) mothers of children with IgE-mediated food allergies. Reported reactions to BM were scored by an allergist, provided only with the details of the possible reaction and not the suspect allergen or route of exposure, as to the likelihood that the reaction was IgE mediated.ResultsA total of 133 mothers completed the survey. After food allergy diagnosis, 47.4% (n = 63) of the mothers reported that they were advised by their health care provider to continue BF without dietary restriction, 17.3% (n = 23) were advised to avoid eating the food(s) their child has allergy to while BF, and in 28.6% (n = 38), this concern was not addressed. A few of the mothers (12%, 16/133) reported that their child experienced an allergic reaction to BM. An allergist evaluated most of these reactions (75%, 12/16) as not likely IgE mediated.ConclusionThis study exposed inconsistent recommendations for mothers providing BM to children with IgE-mediated food allergies. Most mothers were able to consume the food their child has allergy to without adverse sequelae. Standardized, evidence-based recommendations would enhance the well-being of these mother-infant dyads.     

Alterations of lymphocyte subpopulations and TGF-β in children with transient or persistent cow’s milk allergy

The aim of the study was to evaluate changes in surface receptor expression of B and T lymphocytes and concentration of TGF-β in children who either developed tolerance to cow’s milk protein (CMP) or manifested persistent cow’s milk allergy (CMA). The study involved 30 patients with CMA who underwent an open food challenge after 12 months of milk-free diet. After the milk challenge, decreased concentration of CD19⁺CD23+ was observed in children who acquired tolerance to CMP, in comparison with the test before cow’s milk (CM) challenge (42.2% vs. 29.1%, p?=?.006). The same group demonstrated lower concentration of TGF-β than patients with persistent allergy (median 37.9 pg/ml vs. 52.8 pg/ml, p?=?.003, respectively). Moreover, before CM challenge, higher percentage of CD3+CD8+CD28+CD152+ cells (median 2.88% vs. 1.2%, p?=?.03) and CD3+CD4⁺CD25⁺CD62L⁺ (median 42.3% vs. 13.4%, p?=?.032) was noted in children who acquired tolerance to CMP, in comparison with subjects who remained allergic to CMP.     

Interleukin 10 (IL10) and transforming growth factor β1 (TGFβ1) gene polymorphisms in persistent IgE-mediated cow's milk allergy

OBJECTIVES:

The aim of this cross-sectional study was to evaluate whether interleukin 10 (IL10) and transforming growth factor β1 (TGFβ1) gene polymorphisms were associated with persistent IgE-mediated cow''s milk allergy in 50 Brazilian children. The diagnostic criteria were anaphylaxis triggered by cow''s milk or a positive double-blind, placebo-controlled food challenge. Tolerance was defined as the absence of a clinical response to a double-blind, placebo-controlled food challenge or cow''s milk exposure.

METHOD:

The genomic DNA of the 50 patients and 224 healthy controls (HCs) was used to investigate five IL10 gene polymorphisms (-3575A/T, -2849A/G, -2763A/C, -1082G/A, -592C/A) and one TGFβ1 polymorphism (-509C/T).

RESULTS:

Among the five IL10 polymorphisms analyzed, homozygosis for the G allele at the -1082 position was significantly higher in the patients compared with the healthy controls (p = 0.027) and in the persistent cow''s milk allergy group compared with the healthy controls (p = 0.001).

CONCLUSIONS:

Homozygosis for the G allele at the IL10 -1082G/A polymorphism is associated with the persistent form of cow''s milk allergy.     

Expression of bone-morphogenetic protein 2 and tumor necrosis factor α correlates with bone metastases in bladder urothelial carcinoma

We have investigated the role of bone-morphogenetic protein (BMP) 2 and tumor necrosis factor α (TNF-α) in 33 patients with bladder cancer (BCa) with bone metastasis. Thirty nonmetastatic BCas were included as controls. Immunohistochemical staining with BMP-2 and TNF-α was performed. Expressions of the factors were quantified and studied statistically. As a result, a trend showing higher expression of BMP-2 and TNF-α was associated with advanced disease. Expressions of BMP-2 and TNF-α were significantly higher in BCa with bone metastases (P = .0002 and P = .0172, respectively). The expression of BMP-2 and TNF-α showed a direct correlation in metastatic and muscle-invasive cases (P = .0202 and P = .0004, respectively) but not in nonmetastatic or noninvasive BCa (P = .1834 and P = .9215, respectively). It is postulated that BMP-2 can be responsible for the mechanism involved in triggering bone metastasis in BCa. The correlation with TNF-α indicates that the interaction of the 2 factors may promote local invasion and distant metastasis, especially to bone.     

Does dystonia always include co-contraction? A study of unconstrained reaching in children with primary and secondary dystonia

Dystonia is a movement disorder in which involuntary or intermittent muscle contractions cause twisting and repetitive movements, abnormal postures, or both. Excessive co-contraction and abnormalities in the time course of reciprocal inhibition between antagonist groups of muscles are considered to be cardinal features of some types of dystonia and reduced speed of movement is often attributed to involuntary activation of antagonist muscles about a joint. In the present study we describe muscle activity during unconstrained multi-joint reaching movements. Children diagnosed with arm dystonia due to cerebral palsy (CP) or primary dystonia (n = 7, 4–16 years, 4 with CP, 3 primary) and similar age healthy subjects pointed alternately to two targets as fast as possible. The children with dystonia showed decreased speed, greater variability, and pauses at targets compared with controls. Decreased speed was mostly due to difficulty in reversing reaching direction, and increased variability was associated with large fluctuations in the duration of the pauses at targets, rather than with variations in the flexion/extension velocity profiles. Surface electromyographic (EMG) activities were examined to assess if the abnormalities observed in the children with dystonia could be explained in terms of increased levels of co-contraction. Unexpectedly, we found that the children with dystonia showed lower levels of co-contraction than the controls during movement, and the pauses at targets were associated with reduced levels of activation rather than with excessive activity in antagonist groups of muscles. Therefore reduced speed of movement during unconstrained reaching may not be due to involuntary activation of the antagonist muscle, and co-contraction of opposing muscles about a joint is not an obligatory feature of multi-joint movement in children with dystonia.     

Distinct expression of interleukin 17, tumor necrosis factor α, transforming growth factor β, and forkhead box P3 in acute rejection after kidney transplantation

The kidney transplant is the main therapeutic alternative for end-stage kidney disease, and rejection is a major complication. The expression of proinflammatory cytokines is related to graft loss, whereas anti-inflammatory cytokines are associated with graft protection. The objective of this study is to evaluate the “in situ” expression of cytokines T helper 1 (tumor necrosis factor α [TNF-α]), T helper 17 (interleukin 17 [IL-17]), and regulatory T cell (transforming growth factor β [TGF-β]) and the expression of forkhead box P3 (FoxP3) in allograft kidney. We evaluated in situ expression of cytokines in allograft kidney under rejection process by indirect immunohistochemistry. Eighteen renal graft biopsies were from patients with episodes of rejection. The in situ expression of IL-17, TNF-α, and TGF-β was significantly higher in patients with acute rejection when compared with the control group. In contrast, analysis of FoxP3 expression showed few positive cells in patients with acute rejection compared with the control group. The results suggest that the expression of proinflammatory cytokines (IL-17 and TNF-α) contributes to the mechanisms of kidney transplant rejection. The increase in TGF-β expression might be an attempt to establish a process of immunoregulation or even to induce higher production of IL-17. The last hypothesis is supported by the observation of a reduced expression of FoxP3 and elevated levels of IL-17.     

Functional and morphological effects of tumour necrosis factor α in an interleukin 6-producing pulmonary large cell carcinoma with sarcomatoid features

We established a clonal cell line, HAT.MC8, derived from a human pulmonary large cell carcinoma with sarcomatoid features. This cell line was successfully maintained in a protein-free medium and exhibited sarcomatoid fibroblastic features in vitro. The cells constitutively produced a large amount of interleukin 6 (IL-6) in vitro. Tumour necrosis factor (TNF-) not only stimulated HAT.MC8 cells to produce IL-6, but also induced a morphological change from sarcomatoid fibroblastic to epithelial features. Although this change was related to actin and zonula adherens, there was no evidence that E-cadherin participated in the change. Interleukin 1 (IL-1) had a stimulatory effect on IL-6 production by HAT.MC8 cells, but no influence on the morphology of the cells.     

Single molecule measurements of tumor necrosis factor α and interleukin-6 in the plasma of patients with Crohn's disease

The quantitative measurement of inflammatory cytokines in blood has been limited by insufficient sensitivity of conventional immunoassays. This limitation has prevented the widespread clinical monitoring of cytokine concentrations in chronic inflammatory diseases. We applied a sensitive, single molecule detection technology to measure TNF-α and IL-6 in the plasma of patients with Crohn's disease (CD), before and after treatment with anti-TNF-α therapy. Plasma from 17 patients with CD was collected prior to initiation of anti-TNF-α therapy, and the Crohn's disease activity index (CDAI) was determined for each patient. A sub-set of these patients returned for follow up 12 weeks after treatment started. Plasma from age- and gender-matched controls was also collected. Digital ELISAs were developed for TNF-α and IL-6, and the plasma concentrations of these cytokines were determined using digital ELISA. The limits of detection of the TNF-α and IL-6 digital ELISAs were 0.008 pg/mL and 0.006 pg/mL, respectively. Both cytokines were detected in all samples using digital ELISA and the concentrations of TNF-α and IL-6 in the plasma of patients with CD were (3.6±0.9) pg/mL and (10.9±11.2) pg/mL, respectively. TNF-α levels in patients and healthy controls were not significantly different, but the IL-6 levels in plasma were significantly elevated in patients compared to controls. After therapy, the mean reduction of the concentrations of free TNF-α and IL-6 were 46% and 58%, respectively. Digital ELISA provided the first quantitative measurements of TNF-α and IL-6 concentrations in the plasma of all patients in a population with CD. The changes in cytokine concentrations after therapy--which could be quantified because of the high sensitivity of digital ELISA--could be used for monitoring therapeutic efficacy.     

Threshold serum concentrations of tumour necrosis factor alpha (TNFα) as a potential marker of the presence of microangiopathy in children and adolescents with type 1 diabetes mellitus (T1DM)

The aim of this study was to estimate the threshold serum concentrations of advanced glycation end products (AGEs) and their soluble receptors (sRAGE) as well as tumour necrosis factor alpha (TNFα), vascular endothelial growth factor₁₆₅ (VEGF₁₆₅) and interleukin-12 (IL-12) in predicting the occurrence of microangiopathy in children and adolescents with type 1 diabetes mellitus (T1DM). We studied 88 children and adolescents (age range: 6–20 yrs) with T1DM and 32 control subjects (age range: 7–20 yrs). All study participants had their daily urinary albumin excretion, HbA1c and serum creatinine levels measured, and underwent an eye examination and 24-h blood pressure monitoring. Moreover, serum concentrations of AGEs, sRAGE, TNFα, VEGF₁₆₅ and IL-12 were measured. In order to calculate the threshold values of the studied parameters, the Receiver Operating Characteristic (ROC) curve analysis was used. The results of our study have shown that among all the studied parameters a discriminative ability was found for TNFα, VEGF₁₆₅, duration of the disease, serum AGEs concentrations and daily urinary albumin excretion. However, the highest value of the area under the ROC curve (AUC_ROC) in predicting the occurrence of diabetic microangiopathy was found for serum TNFα concentrations with its threshold value of 1.7 pg/ml [AUC_ROC = 0.88 (95% CI: 0.79–0.97)]. The sensitivity and specificity for this variable was at the level of 85.7% and 94.3%, respectively. In conclusion, according to our results serum TNFα concentrations over 1.7 pg/ml may point to the presence of diabetic microangiopathy in children and adolescents T1DM.     

B-cell activating factor of the tumour necrosis factor family expression in blood monocytes and T cells from patients with primary Sjögren's syndrome

We investigated B-cell activating factor of the tumour necrosis factor family (BAFF) level in peripheral blood mononuclear cells (PBMCs), monocytes and T cells from patients with primary Sjögren's syndrome (pSS) and controls both ex vivo and in vitro after cytokine stimulation. PBMCs, monocytes and T cells were isolated from 15 patients with pSS and 17 controls. Cells were cultured alone or with interferon (IFN)α, IFNγ and interleukin 10 (IL-10). T cells were stimulated with phytohaemagglutin and anti-CD3. BAFF protein was assessed by enzyme-linked immunosorbent assay. Ex vivo , no difference was observed in BAFF mRNA level in PBMCs and monocytes from patients and controls. Blood monocytes were the main cell type secreting BAFF both in patients and controls. In vitro , after IFNα stimulation, BAFF mRNA level was significantly higher in cells from patients than from controls (63.8 versus 20.7, P  = 0.03). T cells from patients secreted a higher level of BAFF protein than those from healthy donor cells (17.4 versus 2.9 pg/ml, respectively, P  = 0.04) but at a lower level than that from monocytes. Stimulation of T cells did not change BAFF secretion level. The induction of Th17 cells showed no increased BAFF expression. In conclusion, similar to epithelial cells, blood monocytes in patients with pSS show increased production of BAFF under IFNα, which confirms the involvement of IFNα in pSS. BAFF expression is also increased in blood T cells of such patients, independently of T-cell stimulation.     

Association between degree of bone-erosion and synovial fluid-levels of tumor necrosis factor α in the knee-joints of patients with rheumatoid arthritis

Objective To determine whether concentrations of cytokines and matrix-degrading enzymes in synovial fluid of patients with rheumatoid arthritis or osteoarthritis are associated with the degree of bone-destruction in the same joint.Methods Determination of Interleukin-1, IL-1, IL-1-receptor-antagonist, IL-6, IL-8, tumor necrosis factor (by ELISA), collagenase-activity and caseinase-activity (by substrate-assays) in the SF (knee) of patients with RA (n42) or OA (n35). The degree of bone-destruction was assessed radiographically.Results SF cytokine- and enzyme-levels were higher in patients with RA than in those with OA. In the RA group, SF-levels of TNF were positively correlated with the degree of bone destruction of the respective joint. No correlation was found between radiographically assessed joint changes and SF-concentrations of other cytokines, enzyme activities, serum CRP, or duration of disease. In the OA-group, none of the examined parameters was associated with the degree of joint destruction.Conclusions Our data may support the assumption of TNF playing an important role in joint destruction in RA. Possible alternative conclusions are discussed.     

Serum concentrations of adiponectin,leptin, resistin,ghrelin and insulin and their association with obesity indices in obese normo- and hypertensive patients – pilot study

Introduction

Hypertension often coexists with obesity. Adipokines, ghrelin and insulin play important roles in the pathogenesis of both diseases. The aim of this study was to compare adiponectin, leptin, resistin, insulin and ghrelin mean serum concentrations and insulin resistance (HOMA-IR) in normo- and hypertensive patients with obesity.

Material and methods

All included patients were divided on the following groups: non-diabetic hypertensive patients with class I obesity (group A, n = 21) and class II/III obesity (group B, n = 10), and normotensive obese (class I)patients (group C, n = 7). Correlations between obesity indices (body mass index [BMI], waist-to-hip ratio [WHR], waist circumference [WC]), HOMA-IR, and hormone and adipokine serum levels were also analyzed.

Results

Leptin level and HOMA-IR were significantly higher in group B compared to group C (9.74 ±3.88 ng/ml vs. 4.53 ±3.00 ng/ml; p < 0.02 and 3.30 ±1.59 vs. 1.65 ±0.41; p < 0.02, respectively). A negative correlation between WC and adiponectin level (R = –0.6275; p < 0.01) and a positive correlation between WC and insulin concentration (R = 0.5122; p< 0.05) as well as with HOMA-IR (R = 0.5228; p < 0.02) were found in group A. Negative correlations between BMI and ghrelin level (R = –0.7052; p < 0.05), WHR and adiponectin level (R = –0.6912; p < 0.05) and WHR and leptin level (R = –0.6728; p < 0.05) were observed in group B.

Conclusions

Insulin resistance and leptin may be important pathogenic factors in hypertensive patients with severe obesity. Indices of abdominal obesity (WC, WHR) correlate better than BMI with HOMA-IR, insulin, adiponectin and leptin serum levels in hypertensive obese patients.     

Associations between cytotoxic T lymphocyte-associated antigen-4 polymorphisms and serum tumor necrosis factor-α and interferon-γ levels in patients with chronic hepatitis B virus infection

Background  

Cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) polymorphism, which may affect host immune response including cytokines production, is thought to be associated with hepatitis B virus (HBV) infection. This study investigated the associations between CTLA4 polymorphism and serum tumor necrosis factor (TNF)-α and interferon (IFN)-γ levels in patients with chronic HBV infection.     

Morphology and immuno-distribution of the histamine H4 receptor and histamine – releasing factor in choroid plexus of patients with paraneoplastic cerebellar degeneration

Objective and Design:  The study was aimed at screening out the mimetic peptides from the binding site of lipopolysaccharide binding protein and CD 14, and then observing if the mimetic peptide will inhibit in vitro LPS-induced inflammatory reaction and function as an anti-endotoxin in the model of LPS-induced acute lung injury. Material and Methods:  Human monocytic cell line (U937) was used in vitro. Thirty three-month-old SD rats were used. Phage display peptide library was adapted to screen mimetic peptide sequences. Treatment:  U937 cells were exposed to treatment with LPS and rhLBP and then were incubated with MP12 at three different concentrations after they were induced and differentiated by PMA. LPS intravenous injection was used to establish a model of rat acute lung injury which was later treated with intravenous injection of MP12. Results:  We successfully obtained the mimetic peptide of lipopolysaccharide-binding protein and CD 14 binding site, the gene sequence of which is FHRWPTWPLPSP (MP12). MP12 can markedly inhibit LPS induced TNF-α expression. MP12 can evidently increase PaO₂ of rats with acute lung injury and also increase the survival rate of these rats. Conclusions:  MP12 (FHRWPTWPLPSP) has the same function as mimetic of lipopolysaccharide-binding protein and CD 14 binding site. The application of MP12, both in vitro and in vivo, confers the biological activity required to antagonise LBP/CD14 and block LPS inflammatory signals, and it can markedly enhance PaO₂ of rats suffering from acute lung injury and also enhance their survival rate. Received 28 April 2008; returned for revision 2 June 2008; received from final revision 4 October 2008; accepted by A. Falus 7 October 2008