Distributions of interleukin-6 (IL-6) promoter and metallothionein 2A (MT2A) core promoter region gene polymorphisms and their associations with aging in Turkish population

Aging is determined as the product of an interaction among genetic, environmental and lifestyle factors. As interleukin-6 (IL-6) and metallothioneins (MTs) are related to inflammation and oxidative stress response, their genes are appropriate candidate for aging, age related diseases and infections. The aim of this study was to investigate the association between the IL-6 −174 G/C promoter region and MT2A −5 A/G core promoter region single nucleotide polymorphisms (SNPs) with longevity in Turkish population. Blood samples were collected from 354 individuals between 18 and 95 years of age. Individuals were classified into four groups according to their ages as 20–40, 41–60, 61–80, >80. IL-6 and MT2A polymorphisms were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Mean ages of individuals with IL-6 −74 C⁻ carriers and C⁺ carriers were 49.82 ± 20.45 years and 59.82 ± 16.82 years, respectively. For the MT2A polymorphism, mean ages were estimated as 56.18 ± 19.50 years for G⁻ carriers and 47.59 ± 13.45 years for G⁺ carriers. As a result, when the IL-6 and MT2A polymorphisms were compared with the mean ages and age groups, statistically significant associations were found (p < 0.001 and p < 0.05, respectively). In conclusion, these data support that the IL-6 −174 C⁺ carriers and MT2A −5 G⁻ carriers may be more advantageous for longevity.     

The effect of royal jelly on CD3+, CD5+, CD45+ T-cell and CD68+ cell distribution in the colon of rats with acetic acid-induced colitis

BackgroundTraditional medicines and health supplements have historically been used to treat many illnesses but most of them have not been evaluated objectively to prove their efficacy. We have been investigating the effects of royal jelly (RJ) supplements on acetic acid-induced colitis on the distribution of CD3⁺, CD5⁺, CD45⁺ T-cell and CD68⁺ cells in rats.MethodsThe rats were divided into four equal groups: control group, royal jelly-treated (RJ – 150 mg kg^?1 body weight), acetic acid-treated (colitis) and acetic acid-treated (colitis) + royal jelly (CRJ – 150 mg kg^?1 body weight). Colitis was induced by intracolonic instillation of 4% acetic acid; the control group received physiological saline (10 mL kg^?1). Colon samples were obtained under deep anaesthesia from animals in four groups. Tissues were fixed in 10% formalin neutral buffer solution for 24 h and embedded in paraffin.ResultsThe proliferative response of CD3⁺ and CD45⁺ T cells stimulated with colitis was affected by colitis treated with RJ. No differences were found in CD5⁺ T cells and CD68⁺ macrophages in the colitis treated with RJ.ConclusionsThis study has shown that RJ has anti-inflammatory and cell regeneration effect in the colon of rats with acetic acid induced colitis.     

Determination of urine tumor necrosis factor,IL-6, IL-8, and serum IL-6 in patients with hemorrhagic fever with renal syndrome

ObjectiveThe aim of this study was to explore the role of cytokines in the pathogenesis of hemorrhagic fever with renal syndrome (HFRS).MethodsDouble-antibody sandwich ELISA was used to determine serum interleukin (IL)-6, urine tumor necrosis factor (TNF), IL-6, and IL-8 levels in 56 patients with HFRS.ResultsSerum IL-6, urine TNF, IL-6, and IL-8 concentrations in HFRS patients were significantly higher than those in the control group (p < 0.001). The concentrations increased at fever stage, then continued to increase during the hypotension stage and peaked at the oliguria stage. The concentrations of serum IL-6, urine TNF, IL-6, and IL-8 increased according to the severity of the disease, and differed greatly among different types of the disease. Serum IL-6 had remarkable relationships with serum specific antibodies. It was positively related to serum β2-microglobulin (β2-MG), blood ureanitrogen (BUN), and creatinine (Cr). Significant positive relationships were also found both between urine IL-6 and TNF, and between IL-6 and IL-8 (r = 0.5768, p < 0.05; r = 0.3760, p < 0.01).ConclusionTNF, IL-6, and IL-8 were activated during the course of the disease. IL-6 was associated with the immunopathological lesions caused by the hyperfunction of the humoral immune response. IL-6, IL-8 and TNF were involved in renal immune impairment. Determining them might, to a certain extent, be useful in predicting the prognosis and outcome of patients with HFRS.     

Efficacy of four different moxifloxacin-based triple therapies for first-line H. pylori treatment

Moxifloxacin has been used in the first-line treatment of Helicobacter pylori infection. The optimal dosage and duration have not been assessed.AimTo evaluate the effectiveness of moxifloxacin, amoxicillin and esomeprazole in four regimens, in previously untreated patients infected by H. pylori.Methods and patientsPatients were randomly assigned to: esomeprazole 20 mg b.i.d., amoxicillin 1 g b.i.d., and one of each of the four following dosages of moxifloxacin: moxifloxacin 400 mg b.i.d. for 10 days (EAM800 × 10), moxifloxacin 400 mg b.i.d. for 7 days (EAM800 × 7), moxifloxacin 400 mg b.i.d. for 5 days (EAM800 × 5), moxifloxacin 400 mg o.i.d. for 10 days (EAM400 × 10). Eradication was assessed by the Urea Breath Test (UBT) 2 months following the end of therapy.ResultsNinety-four, 102, 92 and 105 patients were recruited in EAM800 × 10, EAM800 × 7, EAM800 × 5, and EAM400 × 10 respectively. The eradication rate was for Intention-To-Treat (ITT) and Per Protocol (PP) analyses: EAM800 × 10 group ITT: 90.4%, PP: 94.4%; EAM800 × 7 group ITT: 80.3%, PP: 86.3%; EAM800 × 5 group ITT: 71.4%, PP: 75.2%; EAM400 × 10 group ITT: 80.0%, PP 84.8%. A statistically significant difference was reached between EAM800 × 10 vs. EAM800 × 7 (ITT and PP: P < 0.05), and between EAM800 × 10 vs. EAM800 × 5 (ITT and PP: P < 0.01) and vs. EAM400 × 10 (ITT: P < 0.05; PP: P < 0.04). Thirty patients treated unsuccessfully with EAM800 × 5 and EAM400 × 10 were re-treated with EAM800 × 10 with an eradication rate of 86.7% (ITT) and 92.2% (PP). Nineteen patients with positive UBT after EAM800 × 10 and EAM800 × 7 underwent a second-line rifabutin-based therapy with an eradication rate of 84.2% (ITT and PP).ConclusionA triple therapy with 800 mg of moxifloxacin a day for 10 days is more effective than the same treatment for 5 or 7 days and a treatment with 400 mg of moxifloxacin a day for 10 days for the first-line eradication of H. pylori infection. The high cost of moxifloxacin-based treatment however, may limit its wide use as first-line treatment of H. pylori infection.     

IFN-γ stimulation of dental follicle mesenchymal stem cells modulates immune response of CD4+ T lymphocytes in Der p1+ asthmatic patients in vitro

BackgroundHouse dust mite (Dermataphagoides pteronyssinus) is a widespread risk factor in the development of asthma. CD4⁺ T lymphocytes have an important role in the pathogenesis of allergic asthma by polarizing to Th2 cells.ObjectiveWe aimed to evaluate the immunoregulatory effects of dental follicle mesenchymal stem cells with and without IFN-γ stimulation on peripheral blood mononuclear cells of house dust mite sensitive asthmatic patients, and compared those with Dexamethasone as a systemic steroid.Material and methodsPBMC of asthmatic patients and healthy individuals separately cultured with or without DF-MSCs in the presence and absence of IFN-γ or Der p1 or Dexamethasone for 72 h. CD4⁺ T proliferation, cell viability, CD4⁺CD25⁺FoxP3⁺ Treg cell frequency and cytokine profiles of PBMC were evaluated via flow cytometry.ResultsDF-MSCs suppressed proliferation of CD4⁺ T lymphocytes (p_CDmix < 0.01, p_Derp1 < 0.01, p_IFN < 0.005) by increasing the number of FoxP3 expressing CD4⁺CD25⁺ T regulatory cells (p_CDmix < 0.005, p_Derp1 < 0.01, p_IFN < 0.001) and suppressed lymphocyte apoptosis (p_CDmix < 0.05, p_Derp1 < 0.05, p_IFN < 0.05), while Dexamethasone increased the apoptosis and decreased Treg cell frequency in asthmatic patients. IFN-γ stimulation increased the suppressive effect of DF-MSCs and also enhanced the frequency of FoxP3 expressing CD4⁺CD25⁺ T regulatory cells. The cytokine levels were regulated by DF-MSCs by reducing IL-4 cytokine levels (p_CDmix < 0.01, p_Derp1 < 0.05, p_IFN < 0.05) and upregulating IFN-γ levels (p_CDmix < 0.01, p_Derp1 < 0.05, p_IFN < 0.005) in asthmatic patients.ConclusionIFN-γ stimulated DF-MSCs were found to have a high modulatory effect on CD4⁺ T cell responses, while Dexamethasone had an apoptotic effect on CD4⁺ T cells in asthmatic patients. DF-MSCs may be a new cell-based therapy option for allergic diseases including asthma.     

IL-17 in cutaneous lupus erythematosus

BackgroundLupus erythematosus (LE) is a heterogeneous disease with broad clinical spectrum from cutaneous to visceral and systemic inflammation. IL-17 isoforms (IL-17A and IL-17F) are proinflammatory cytokines with unclear implications in lupus erythematosus pathogenesis. In this study we focused upon IL-17 in normal and modified lupus skin with a correlative study between local and serological expression.Material and methods89 subjects were recruited and divided in 5 groups—10 patients with psoriasis (disease control group), 13 healthy controls, 26 with discoid chronic lupus (DLE), 23 with systemic lupus erythematosus (SLE) and 17 with subacute lupus erythematosus (SCLE). Blood samples and skin punched-biopsy specimens were performed. Serum IL-17A, IL-17F, and IL-23 concentrations were determined by ELISA. Skin IL-17A and CD4 expression were evaluated by immunohistochemistry.ResultsImmunohistochemical expression of IL-17A was higher in DLE, SCLE and SLE patients than in negative control subjects (all p < 0.05). Serum IL-17A concentrations were higher in DLE and SLE patients than in negative controls (p < 0.05). Serum IL-17A levels were similar in SCLE and negative controls (p > 0.05). Serum IL-17F concentrations were higher in DLE, SCLE and SLE patients than in healthy controls (all p < 0.05). In DLE, SCLE, SLE patients and healthy controls we observed comparable levels of IL-23 (p > 0.05). Serum anti Ro antibodies correlate with IL-17A+ lymphocytes from SCLE lesion and SLE normal skin (all p < 0.05).ConclusionIL-17 isoforms (IL-17A and IL-17F) are implicated in SLE but also in DLE and SCLE immunopathogenesis.     

Role of Inflammasome Activation in Systemic Lupus Erythematosus: Are Innate Immune Cells Activated?

BackgroundSystemic lupus erythematosus (SLE) is characterized by a wide spectrum of clinical and immunological abnormalities. New data have emerged about the role of inflammasomes in autoimmune diseases. We aimed to investigate whether basal inflammasome activation occurs in SLE patients, and whether a relationship between inflammasome-related-cytokines and disease activity exists.MethodsFourteen (14) consecutive SLE patients and 13 healthy individuals, matched by sex, age and ethnicity, were included. Demographics, laboratory and clinical data were recorded. Peripheral blood mononuclear cells (PBMCs) from patients and controls were obtained and monocytes were isolated by negative selection. Purified monocytes were stimulated with LPS in the presence or absence of Caspase-1 inhibitor. CD14 and Caspase-1 expression were analyzed by flow cytometry. Cytokine levels were determined in plasma and culture supernatants by ELISA. Student's t test and Mann–Whitney tests were used for statistical analysis.ResultsThe percentage of CD14⁺/Caspase-1⁺ was significantly higher in monocytes from SLE patients compared to normal controls (p < 0.01). These findings paralleled with higher plasma levels of IL-1β (p < 0.05) and IL-18 (p < 0.01) in those patients. Purified monocytes from SLE patients displayed a robust inflammatory response after LPS stimulation where Caspase-1, IL-1β and IL-18 were highly expressed. Plasma levels of IL-18 were also significantly higher in SLE patients with active disease (p < 0.05). In addition, the production of IL-18 was reduced by 3 fold when Caspase-1 inhibitor was added to the cultures.ConclusionsMonocytes from SLE patients exhibited increased inflammasome activation, characterized by high expression of Caspase-1, IL-1β and IL-18. Caspase-1 specific inhibitor decreased inflammasome activation (in vitro) by suppressing the production of IL-18.     

Th-17 cells and serum IL-17 in rheumatoid arthritis patients: Correlation with disease activity and severity

Aim of the workTo investigate the role of T-helper 17 (Th17) cells in peripheral blood and serum interleukin-17 (IL-17) in rheumatoid arthritis (RA) patients, and their correlation with disease activity and joint destruction.Patients and methodsThis study included forty RA patients and twenty matched healthy controls. Disease activity score in 28 joints (DAS-28), erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), rheumatoid factor (RF), anti-CCP, tumor necrosis factor alpha (TNF-α), serum IL-17 and Th17 cells in peripheral blood were measured. Radiological assessment using modified Sharp/van der Heijde (mSvH) score for hand and feet in addition to MRI score for the wrist and metacarpophalangeal (MCP) joints were performed for detection of synovitis and bone erosion.ResultsThe patients were 38 females and 2 males with a mean of 41.15 ± 5.85 years and disease duration of 15.6 ± 4.62 years. Serum IL-17 and Th17 cells in peripheral blood were found to be significantly increased in RA patients (204.1 ± 33.8 pg/ml and 4.62 ± 1.13%) than in controls (25.36 ± 5.39 pg/ml and 0.7 ± 0.021%) (p < 0.001). Th17 cells significantly correlated with serum IL-17 (r 0.88, p < 0.001). Both Th17 cells and serum IL-17 significantly correlated with DAS-28, ESR, CRP, TNF-α, Van der Heijde modification score and MRI scores for wrist and MCP joints for synovitis and bone erosion (all with a p < 0.001).ConclusionThis study demonstrates an important role for Th-17 cells and serum IL-17 in the pathogenesis of the inflammatory and destructive pattern characteristic of RA.     

Impaired calcium-calmodulin-dependent inactivation of Cav1.2 contributes to loss of sarcoplasmic reticulum calcium release refractoriness in mice lacking calsequestrin 2

AimsIn cardiac muscle, Ca^2 + release from sarcoplasmic reticulum (SR) is reduced with successively shorter coupling intervals of premature stimuli, a phenomenon known as SR Ca^2 + release refractoriness. We recently reported that the SR luminal Ca^2 + binding protein calsequestrin 2 (Casq2) contributes to release refractoriness in intact mouse hearts, but the underlying mechanisms remain unclear. Here, we further investigate the mechanisms responsible for physiological release refractoriness.Methods and resultsGene-targeted ablation of Casq2 (Casq2 KO) abolished SR Ca^2 + release refractoriness in isolated mouse ventricular myocytes. Surprisingly, impaired Ca^2 +-dependent inactivation of L-type Ca^2 + current (I_Ca), which is responsible for triggering SR Ca^2 + release, significantly contributed to loss of Ca^2 + release refractoriness in Casq2 KO myocytes. Recovery from Ca^2 +-dependent inactivation of I_Ca was significantly accelerated in Casq2 KO compared to wild-type (WT) myocytes. In contrast, voltage-dependent inactivation measured by using Ba^2 + as charge carrier was not significantly different between WT and Casq2 KO myocytes. Ca^2 +-dependent inactivation of I_Ca was normalized by intracellular dialysis of excess apo-CaM (20 μM), which also partially restored physiological Ca^2 + release refractoriness in Casq2 KO myocytes.ConclusionsOur findings reveal that the intra-SR protein Casq2 is largely responsible for the phenomenon of SR Ca^2 + release refractoriness in murine ventricular myocytes. We also report a novel mechanism of impaired Ca^2 +-CaM-dependent inactivation of Ca_v1.2, which contributes to the loss of SR Ca^2 + release refractoriness in the Casq2 KO mouse model and, therefore, may further increase risk for ventricular arrhythmia in vivo.     

IGF-1 alters the human parietal pleural electrochemical profile by inhibiting ion trans-cellular transportation after interaction with its receptor

ObjectiveThe effect of IGF-1 in the human pleural permeability and the underlying mechanisms involved were investigated.DesignSpecimens from thoracic surgical patients were mounted in Ussing chambers. Solutions containing IGF-1 (1 nM–100 nM) and IGF-1 Receptor Inhibitor (1 μΜ), amiloride 10 μM (Na⁺ channel blocker) and ouabain 1 mM (Na⁺–K⁺ pump inhibitor) were used in order to investigate receptor and ion transporter involvement respectively. Trans-mesothelial Resistance (R_TM) across the pleural membrane was determined as a permeability indicator. Immunohistochemistry for IGF-1 receptors was performed.ResultsIGF-1 increased R_TM when added on the interstitial surface for all concentrations (p = .008, 1 nM–100 nM) and decreased it on the mesothelial surface for higher concentrations (p = .046, 100 nM). Amiloride and ouabain inhibited this effect. The IGF-1 Receptor Inhibitor also totally inhibited this effect. Immonuhistochemistry demonstrated the presence of IGF-1 receptors in the pleura.ConclusionsIt is concluded that IGF-1 changes the electrophysiology of the human parietal pleura by hindering the normal ion transportation and therefore the pleural fluid recycling process. This event is achieved after IGF-1 interaction with its receptor which is present in the human pleura.     

Expression of T-cell KV1.3 potassium channel correlates with pro-inflammatory cytokines and disease activity in ulcerative colitis

Background and aimsPotassium channels, K_V1.3 and K_Ca3.1, have been suggested to control T-cell activation, proliferation, and cytokine production and may thus constitute targets for anti-inflammatory therapy. Ulcerative colitis (UC) is a chronic inflammatory bowel disease characterized by excessive T-cell infiltration and cytokine production. It is unknown if K_V1.3 and K_Ca3.1 in the inflamed mucosa are markers of active UC. We hypothesized that K_V1.3 and K_Ca3.1 correlate with disease activity and cytokine production in patients with UC.MethodsMucosal biopsies were collected from patients with active UC (n = 33) and controls (n = 15). Protein and mRNA expression of K_V1.3 and K_Ca3.1, immune cell markers, and pro-inflammatory cytokines were determined by quantitative-real-time-polymerase-chain-reaction (qPCR) and immunofluorescence, and correlated with clinical parameters of inflammation. In-vitro cytokine production was measured in human CD3⁺ T-cells after pharmacological blockade of K_V1.3 and K_Ca3.1.ResultsActive UC K_V1.3 mRNA expression was increased 5-fold compared to controls. Immunofluorescence analyses revealed that K_V1.3 protein was present in inflamed mucosa in 57% of CD4⁺ and 23% of CD8⁺ T-cells. K_V1.3 was virtually absent on infiltrating macrophages. K_V1.3 mRNA expression correlated significantly with mRNA expression of pro-inflammatory cytokines TNF-α (R² = 0.61) and IL-17A (R² = 0.51), the mayo endoscopic subscore (R² = 0.13), and histological inflammation (R² = 0.23). In-vitro blockade of T-cell K_V1.3 and K_Ca3.1 decreased production of IFN-γ, TNF-α, and IL-17A.ConclusionsHigh levels of K_V1.3 in CD4 and CD8 positive T-cells infiltrates are associated with production of pro-inflammatory IL-17A and TNF-α in active UC. K_V1.3 may serve as a marker of disease activity and pharmacological blockade might constitute a novel immunosuppressive strategy.     

IL-17 is a key cytokine correlating with disease activity and clinical presentation of systemic lupus erythematosus

AimsTo determine the role of IL-17 cytokine in systemic lupus erythematosus (SLE) patients and its association with clinical presentation of the disease and disease activity.Methods72 SLE patients and 70 healthy age and sex matched controls were included in the study. SLE disease activity was assessed in all patients with SLE disease activity index (SLEDAI-2K) scores. Plasma levels of IL-6, and IL-17 were measured by enzyme-linked immunosorbent assay and correlated their levels with clinical manifestations of the disease and SLEDAI-2K.ResultsPlasma levels of IL-6 and IL-17 were significantly elevated in SLE patients than in control subjects (13.98 ± 6.95 versus 7.47 ± 1.23 pg/mL) and (19.47 ± 10.21 versus 9.93 ± 1.89 pg/mL), respectively. IL-6 and IL-17 were positively correlated with SLEDAI-2K scores (r = 0.684 at P < 0.001, r = 0.322 at P = 0.006), and lupus nephritis (r = 0.364 at P = 0.002, r = 0.474 at P < 0.001) respectively; similarly, the IL-17/IL-6 ratio was positively correlated with SLEDAI-2K (r = 0.243 at P = 0.039). Also, the level of both cytokines was positively correlated to each other during periods of disease activity (r = 0.755, P < 0.001) as well as during remission (r = 0.384, P = 0.040).ConclusionOver-expression of IL-17 correlates with disease activity of SLE. A longitudinal study in a larger cohort of SLE patients can help validate the results.     

Comparison of quantitative measurements between two different intravascular ultrasound systems: In vitro and in vivo studies

BackgroundAlthough intravascular ultrasound (IVUS) allows for precise measurements of coronary artery dimension, variability in quantitative measurements among currently available different IVUS systems is unknown. The aim of study was to compare two different IVUS catheters and consoles to verify their accuracy and compatibility.Methods(1) In vitro study: IVUS imaging was performed in a concentric cylindrical phantom with 6 sections of known, cross-sectional diameter ranging from 3.0 to 8.0 mm. The minimum lumen diameter (MLD) and lumen cross sectional area (CSA) were measured and compared. (2) In vivo study: IVUS imaging was performed in 69 coronary arterial segments from 20 patients. The external elastic membrane cross sectional area (EEM-CSA), lumen CSA, and plaque plus media (P + M) CSA were measured and compared between the two IVUS systems.Results(1) In vitro study: MLD and lumen CSA obtained by the two IVUS systems correlated well with the actual values. (2) In vivo study: EEM-, lumen and P + M CSA obtained by the two IVUS systems showed good correlations (R² = 0.973, p < 0.0001; R² = 0.938, p < 0.0001; R² = 0.949, p < 0.0001, respectively).ConclusionsQuantitative measurements by 2 different, currently available IVUS systems were accurate and comparable. These results suggest that the 2 different IVUS catheters/systems may be alternatively used during clinical studies assessing coronary arterial size.     

Microscopic colitis patients have increased proportions of Ki67+ proliferating and CD45RO+ active/memory CD8+ and CD4+8+ mucosal T cells

BackgroundCollagenous colitis (CC) and lymphocytic colitis (LC) are chronic inflammatory bowel disorders of unknown etiology. This study investigated phenotypic characteristics of the mucosal lymphocytes in CC and LC.MethodsLamina propria and intraepithelial lymphocytes (LPLs, IELs) isolated from mucosal biopsies from CC (n = 7), LC (n = 6), as well as LC or CC patients in histopathological remission, (LC-HR) (n = 6) and CC-HR (n = 4) and non-inflamed controls (n = 10) were phenotypically characterized by four-color flow cytometry.ResultsThe proportions of CD8⁺ IELs were increased in CC and LC (p < 0.01) compared to controls. Increased proportions of CD45RO⁺CD8⁺ IELs and LPLs were observed in LC and even more in CC patients (p < 0.01). Both CC (p < 0.05) and LC patients had elevated proportions of CD4⁺8⁺ IELs and LPLs compared to controls. The proportions of CD45RO⁺ cells were increased in CD4⁺8⁺ IELs and LPLs (p < 0.05) in CC and LC patients compared to controls. Both CC (p < 0.05) and LC patients had higher proportions of Ki67⁺CD8⁺ IELs and LPLs compared to controls.In contrast, decreased proportions of CD4⁺ LPLs were observed in CC and LC as well as CD4⁺ IELs in LC compared to controls. Increased proportions of Ki67⁺CD4⁺ IELs and LPLs (p < 0.05) were observed in CC and LC patients.CC-HR but not LC-HR patients demonstrated normalized proportions of both IELs and LPLs compared to CC and LC patients respectively.ConclusionLC and CC patients have differences in mucosal lymphocyte subsets, with increased proportions of Ki67⁺ and CD45RO⁺ CD8⁺ and CD4⁺8⁺ mucosal T cells.     

Relación entre la expresión de IL-2 e IL-4 y sus polimorfismos y los riesgos de padecer infección por Mycoplasma pneumoniae y asma en niños

IntroductionAsthma is an inflammatory disorder of the airways and the symptoms of asthma could be exacerbated by Mycoplasma pneumoniae infection. Interleukin-2 and interleukin-4 have been implicated in immune and inflammatory reactions. We examined the associations of IL2 and IL4 polymorphisms and expression with the risks of asthma and M. pneumoniae infection in children.Methods392 asthmatic children and 849 controls were recruited into the study. Eight polymorphisms in IL2 and IL4 were genotyped with Sequenom MassARRAY platform. M. pneumoniae infection and copy number was determined with fluorescence PCR. IL-2 and IL-4 serum expression levels were determined by using ELISA.ResultsWe found a significant association of IL2 rs6534349 polymorphism with increased asthma risk (heterozygotes, P = .029; homozygous variants; P = .013) and of IL4 rs2227284 polymorphism with reduced asthma risk (heterozygotes, P = .026; homozygous variants; P = .001). Besides, the association of other polymorphisms, except rs2070874 polymorphism, became apparent when the asthmatic children were grouped according to GINA classification of asthma control and severity. In addition, IL-2 and IL-4 serum expression levels were significantly higher in M. pneumoniae negative (P = .038) and positive (P = .011) subjects respectively. This observation holds true among asthmatic patients (P = .016 for IL-2 and P = .042 for IL-4), but only the IL-4 observation remained correct among non-asthmatic controls (P = .032). We also observed that the rs6534349 GG genotype was significantly associated with increased odds of getting high load M. pneumoniae infection (P = .0376).ConclusionsIL2 and IL4 could be important biomarkers for estimating the risks of asthma and M. pneumoniae infection in children.     

Deficient or abundant but unable to fight? Estimation of circulating FoxP3+ T regulatory cells and their counteracting FoxP3− in rheumatoid arthritis and correlation with disease activity

Aim of the workPersistent inflammation and recurring activity in rheumatoid arthritis (RA) daring a quarterback of T regulatory cells (Tregs) intrigued rheumatologists. Tregs’ most specific marker is the forkhead box P3 (FoxP3) denoting FoxP3⁺ cells as suppressors whereas FoxP3^? as effectors. This study evaluates subset distribution of peripheral blood (PB) CD4⁺CD25⁺ Tregs according to FoxP3 expression in RA to better understand its role in pathogenesis.Patients and methodsIn our observational cross-sectional study PB Tregs from 40 RA patients and 20 age and sex matched healthy controls (HC) were characterized and quantified by flow cytometry. Disease activity was evaluated by DAS28. Patients were divided into: active RA group (ARA) and remission RA group (RRA).ResultsSignificantly higher CD4⁺CD25⁺FoxP3⁺ Tregs were found on comparing RRA, ARA patients and HC (mean 153.25 ± 6.29, 136.3 ± 3.27 and 97.25 ± 6.25, respectively) with a statistically highly significant difference (F = 553.13, p < 0.001). CD4⁺CD25⁺FoxP3^? increased 3-folds in RRA and 4-folds in ARA compared to HC and CD4⁺CD25⁺FoxP3⁺ increased 1.6-folds in RRA and 1.5-folds in ARA. Thus the ratio of FoxP3^?/FoxP3⁺ cells is altered from 1:3 in HC to 2:3 in RRA and 1:1 in ARA. CD4⁺CD25⁺FoxP3⁺ had a highly significant negative correlation with disease activity score DAS28 (r = ?0.86, p < 0.001).ConclusionThough CD4⁺CD25⁺FoxP3⁺ and CD4⁺CD25⁺FoxP3^? are both increased in RA, their balance is more important; abundant FoxP3^? cells with effector function have the upper hand over suppressive FoxP3⁺ cells. This imbalance relates to RA presence and activity. Reinforcing FoxP3 expression might be a good potential therapeutic target in RA.     

CD14++CD16+ monocyte subset expansion in rheumatoid arthritis patients: Relation to disease activity and interleukin-17

Aim of the workMonocytes are divided into three major subsets based on the expression of the cluster of differentiation CD14 and CD16. The aim of this work was to determine which of the CD16⁺ monocyte subpopulations is expanded in rheumatoid arthritis (RA) and its association with disease activity and interleukin-17 (IL17) levels.Patients and methodsFifty-three RA patients and 20 controls were enrolled in this study. Flow cytometry was performed to detect monocyte subsets and IL17 was measured by ELISA. Disease activity score (DAS28) was assessed.ResultsCD14⁺⁺CD16⁺ monocyte percentage was significantly higher in long standing RA patients compared with early patients and controls (p < 0.01, p < 0.001 respectively). It was significantly higher in patients with RA disease activity and remission compared with the controls (p < 0.001, p < 0.01 respectively). It was not significantly associated with resistance to disease modifying antirheumatic drugs (DMARDs), C-reactive protein, rheumatoid factor and anti-CCP positivity (p > 0.05). It significantly correlated with IL17 (p < 0.002). CD14⁺CD16⁺ monocyte percentage was not significantly correlated with any of the above parameters. IL17 level was significantly higher in patients with early and long standing RA compared to controls (p < 0.01, p < 0.001 respectively). IL17 was higher in RA patients with active disease compared to those in remission and controls (p < 0.01, p < 0.001 respectively). It was higher in RA patients resistant to DMARDs than in responding patients (p < 0.017).ConclusionCD14⁺⁺CD16⁺ monocyte subpopulation was expanded in long standing RA and was correlated with IL17 levels indicating its potential pathogenic importance in RA and may represent an attractive target for future therapeutic interventions.     

KB-R7943 restores endothelium-dependent relaxation induced by advanced glycosylation end products in rat aorta

ObjectivesThe aims of this study were to examine the effects of KB-R7943, an inhibitor of Na⁺/Ca^2 + exchanger, on impaired endothelium-dependent relaxation (EDR) induced by advanced glycosylation end products (AGE) in isolated rat aorta.MethodsBoth acetylcholine (ACh)-induced EDR and sodium nitroprusside (SNP)-induced endothelium-independent relaxation (EIR) were measured after the rings were exposed to AGE in the absence and presence of KB-R7943.ResultsCo-incubation of aortic rings with AGE (0.1 g/L) for 24 h resulted in a significant inhibition of EDR, but had no effects on EIR. After incubation of the rings in the co-presence of KB-R7943 (0.1–10 μM) with AGE for 24 h, KB-R7943 (10 μM) significantly attenuated impaired EDR. Superoxide dismutase (200 U/mL) and l-arginine (3 mM) could ameliorate the impairment of EDR caused by AGE, whereas d-arginine (3 mM) had no effect on EDR. Similarly, AGE decreased superoxide dismutase (SOD) activity and the release of nitric oxide (NO), and increased superoxide anion (O₂^.?) production in aortic tissue. KB-R7943 (10 μM) significantly decreased O₂^.? production and increased SOD activity and the NO release.ConclusionsThese results suggest that KB-R7943 attenuated the impairment of EDR elicited by AGE partially through scavenging oxygen free radicals.     

Study of CD4+, CD8+, and natural killer cells (CD16+, CD56+) in children with immune thrombocytopenic purpura

Objective/backgroundTo assess the percentage of CD4⁺, CD8⁺, and natural killer cells (CD16⁺, CD56⁺) in children with immune thrombocytopenic purpura (ITP) at presentation and study their impact on disease chronicity.MethodsThis case–control study was conducted at the Pediatric Hematology and Oncology Unit, Menoufia University Hospital (tertiary care center in Egypt). The study was held on 30 children presenting with ITP; they were followed-up and classified into two groups: 15 children with acute ITP; and 15 children with chronic ITP. Patients were compared to a group of 15 healthy children of matched age and sex. Measurements of CD4⁺, CD8⁺, and natural killer cells (CD16⁺, CD56⁺) by flow cytometry were assessed and compared in these groups.ResultsCD4⁺ and CD4⁺/CD8⁺ were significantly lower in acute and chronic patients than the control group (p < 0.05 and p < 0.001, respectively), with no significant difference between acute and chronic patients (p > 0.05). However, CD8⁺ was significantly higher in acute and chronic patients than the control group (p < 0.05), with no significant difference between acute and chronic patients (p > 0.05). Natural killer cell percent was significantly lower in acute patients than the control group (p < 0.001), with no significant difference between chronic and control groups (p > 0.05).ConclusionITP is associated with immunity dysfunction denoted by the increase in cytotoxic T lymphocytes and the decrease in natural killer cells.