Concentrations Of IFN-gamma Research Articles

Inflammatory Profile of Chronic Rhinosinusitis With Nasal Polyp Patients in Brazil: Multicenter Study.

To determine the inflammatory profile of CRSwNP in Brazil and characterize the subgroups of CRSwNP patients in this population through cluster analysis. Multicenter cross-sectional study involving 15 centers representing different regions of Brazil. Clinical data of 166 patients and 80 controls, aged 18 to 70 years old, number of surgeries for CRS, history of asthma and aspirin sensitivity, and Lund-Mackay scores on CT scans. During nasal endoscopy, we obtained the Lund-Kennedy scores and collected 2 samples of nasal polyps: one for eosinophil and neutrophil tissue counts and one to quantify different cytokines. 79.6% of our patients had 10 or more eosinophils/HPF. CRSwNP groups exhibited significantly lower concentrations of TNF-alpha and significantly higher concentrations of IFN-gamma, CCL11/Eotaxin, CCL24/Eotaxin-2/MPIF-2, and CCL26/Eotaxin-3 versus the control group (Kruskal-Wallis test). Comparison between CRSwNP groups (≥10 vs <10 eosinophils/HPF) showed no difference in cytokine concentration (Mann-Whitney test). Hierarchical clustering and PCA according to cytokine concentrations revealed 2 main Clusters, with a significantly higher concentration of all cytokines in Cluster 1 (n = 35) than in Cluster 2 (n = 121), except IL-6 and IL-33 (Mann-Whitney test). According to ROC curve analysis the best cut-off to differentiate the 2 clusters was 43 eosinophils/HPF. The group with ≥43 presented a higher prevalence of men and a higher Lund-Mackay score (Mann-Whitney test). CRSwNP patients in Brazil present mixed inflammation, with 2 distinct groups (high and low inflammatory pattern) that can be distinguished by tissue eosinophilia of ≥43 eosinophils/HPF cut-off in nasal polyps.

Influence of Different Orthodontic Brackets on Cytokine and Cortisol Profile

Background and Objectives: Orthodontic tooth movement (OTM) requires bone remodeling resulting from complex processes of aseptic inflammation. Recent studies have confirmed close interaction between the immune and skeletal systems. In addition, various orthodontic appliances including fixed systems affect the sublingual microbial composition, and the likelihood of developing inflammatory reactions of the gums is high, especially early in the treatment period. It is known that these systems have both positive and negative effects on the humoral and cellular immune responses. The main aim of the study was to evaluate the influence of self-ligating and conventional brackets on the salivary concentrations of cytokines (IL-6, osteoprotegerin (OPG), TNF-alpha, and IFN-gamma) and cortisol as a marker of stress. Materials and Methods: Forty patients were analyzed at baseline (T0) and 2 months (T2) after fixing self-ligating (Ormco Damond Q) and conventional brackets (Ormco Mini Diamond). Salivary cytokine and cortisol concentrations were evaluated by commercial ELISA kits. Results: Outcomes of our study showed that after two months of treatment with either of these brackets, IFN-gamma and IL-6 levels did not change. However, TNF-alpha decreased with self-ligating brackets (13.36 to 8.32, p = 0.002). The self-ligating bracket system also affects OPG concentration and cortisol levels 2 months after orthodontic activation. The level of OPG in the group of self-ligating brackets decreased significantly (8.55 to 2.72, p = 0.003). Cortisol concentration was significantly higher in the self-ligation group (25.72 to 48.45, p = 0.001) due to the effect of sustained strength movements. Conclusions: Thus, the use of self-ligating and conventional brackets has a different effect on the concentration of cortisol and cytokines (OPG and TNF-alpha) in saliva 2 months after their fixation. Further longitudinal studies are necessary to explore why OPG levels are decreased in case of self-ligating cases and how OPG levels are related to clinical improvement.

The Human Milk Oligosaccharide 2'-Fucosyllactose Shows an Immune-Enhancing Effect in a Cyclophosphamide-Induced Mouse Model.

The 2'-fucosyllactose (2'-FL) is the richest components in a human milk oligosaccharide. Several studies have reported that 2'-FL has beneficial effects in infants. However, there are few studies on its immune-enhancing effects. This research aimed to examine the immune-enhancing effect of 2'-FL on immunosuppression by cyclophosphamide (CCP) in ICR mice. Mice were orally administered distilled water or 0.5 mg/kg B.W. 2'-FL for 14 days. An immunocompromised mouse model was induced using CCP 80 mg/kg B.W. at 12-14 days. Using the CCP had effects on reducing their body weight, organ weight, spleen index, natural killer (NK) cell activity, and cytokines concentration and expression. This study also used concanavalin A-mediated T-cell proliferation to verify the immune-enhancing effects in the sample. Body weight, spleen index, organ weight, and cytokine levels were measured to estimate the immune-enhancing effects. The body weight at 14 days tended to increase, and the spleen weight and index significantly increased in the 2'-FL group compared to the CCP group. The NK cell activity increased in the 2'-FL group compared to the CCP group, but there was no significant difference. The concentration of interleukin (IL)-2 tended to recover in the 2'-FL group compared to the CCP group. The 2'-FL group showed a significant increase of IL-10 and IFN-gamma concentration compared to the CCP group. In addition, there was a trend of increased IL-10 mRNA expression compared to the CCP group. These results revealed that 2'-FL improved CCP-induced immunosuppression, suggesting that 2'-FL may have the potential to enhance the immune system.

In vitro cytotoxicity of human urine and its potential toxic parameters towards bladder cancer cells.

BackgroundBladder cancer (CaB) has a high recurrence rate despite surgery. As bladder is constantly filled with urine, it is worthwhile to investigate whether it could have any detrimental effects on bladder cancer cells.MethodsWe investigated the cytotoxicity of urine samples from CaB patients and normal controls on four CaB cell lines and tested the percentage of cell death, proliferation, adhesion, invasion and colonies formation ability. In order to identify the potential components involving in urine cytotoxicity, we evaluated some basic physiochemical parameters of urines, such as pH, osmolarity, creatinine (Cr), sodium (Na), potassium (K), chloride (Cl), calcium (Ca) and phosphate (PO4). We further compared the pH values of urine samples between CaB who developed recurrence versus those who did not. A more in-depth analysis on inflammatory markers was performed for two representative urine samples which demonstrated opposite cytoxic effects.Results23 CaB patients and 20 normal controls were recruited into this study. According to in vitro experiments, both CaB and non-CaB urines had comparable effect on cell toxicity, proliferation, adhesion, invasion and colonies formation ability in four cell lines, HTB9, RT4, T24 and UMUC3, while RT4 was the most sensitive to urine toxicity. After evaluating the relationship between basic physiochemical parameters and cytotoxicity, we found out that there were strong negative correlations between pH value and 24 hours death rate for the 4 CaB cell lines (HTB9 r = -0.6651, p<0.001; RT4 r = -0.8335, p<0.001; T24 r = -0.4924, p<0.001; UMUC3 r = -0.7066, p<0.001). Osmolarity, urine Cr and PO4 all had weakly or moderately positive correlations with CaB cells on 24 hours death rate. CaB patients who developed recurrence had more alkaline urine than those who did not develop recurrence. In the urine sample with the highest cytoxicity, high concentrations of IL-6 and IFN-gamma were found.ConclusionsOur study confirmed that there was not statistically significant difference in cytotoxicity between CaB and non-CaB urines. However, we identified some parameters that could have an impact on cytotoxicity towards CaB cells. Modifying certain urine characteristics peri-operatively may induce cytotoxicity, avoid tumour re-implantation, and reduce the chance of cancer recurrence.

In:Ge/Cu@calcein polyhedral nanostructure conducted photoelectrochemical biosensor for detection of IFN-gamma

A novel photoelectrochemical (PEC) biosensor based on the In:Ge/Cu@calcein polyhedral nanostructure was fabricated for ultrasensitive determination of interferon (IFN)-gamma. The calcein dye could trigger Cu nano-octahedral to transform Cu nano-polyhedral structure and attach orderly to the its surfaces by electrochemical co-deposition, and facilitate photocurrent response dramatically enhancing by five-folds. The In:Ge/Cu@calcein demonstrates excellent PEC performance resulting from the synergistic sensitization of calcein and In:Ge/Cu nanostructure, as well as supplying rich carboxy groups to couple with other biomolecules for simply fabricating biosensor, which has a wide dynamic range (from 50 fg mL −1 to 1 ng mL −1 concentrations of IFN-gamma), and a lower detection limit of 24 fg mL −1 .

Profiles of interferon-gamma and interleukin-2 in patients after allogeneic hematopoietic stem cell transplantation.

BACKGROUNDAllogeneic hematopoietic stem cell transplantation (allo-HSCT) may be related to the occurrence of complications, including graft-versus-host disease (GvHD) and infections. The pathogenesis of acute GvHD is connected with T lymphocytes, which identify alloantigens on host's antigen-presenting cells, activate production of interferon-gamma (IFN-gamma) and interleukin-2 (IL-2), and act on the immune effector cells and damage tissues and organs.AIMThe aim of the study was to investigate and distinguish serum concentration profiles of IFN-gamma and IL-2 within a 30-d period after allo-HSCT.METHODSWe enrolled 62 patients, i.e., 30 (48%) male and 32 (52%) female subjects [median age 49.5 (19-68) years], after allo-HSCT from siblings (n = 12) or unrelated donors (n = 50) due to acute myeloid leukemia with myeloablative conditioning (n = 26; 42%) and with non-myeloablative conditioning (n = 36; 58%). All patients were given standard immunosuppressive therapy with cyclosporin-A and methotrexate and pre-transplant antithymocyte globulin in the unrelated setting. Blood samples were collected pre-transplant before and after (on day -1) the conditioning therapy and on days +2,+4, +6, +10, +20, and +30 after allo-HSCT. Serum levels of IL-2 and IFN-gamma were determined using ELISA. RESULTSPatients were divided into four groups depending on the presence of acute GvHD and clinical manifestations of infection. Group I included patients with neither acute GvHD nor infections [n = 15 (24%)], group II consisted of patients with infections without acute GvHD [n = 17 (27%)], group III was comprised of patients with acute GvHD without infections [n = 9 (15%)], and group IV included patients with both acute GvHD and infections [n = 21 (34%)]. IFN-gamma concentrations were higher in Group II than in other groups on days +20 (P = 0.014) and +30 (P = 0.008). Post-hoc tests showed lower concentrations of IFN-gamma on day +30 in groups I (P = 0.039) and IV (P = 0.017) compared to group II. The levels of IL-2 were mostly undetectable.CONCLUSIONSerum levels of IFN-gamma following allo-HSCT progressively escalate. High serum levels of IFN-gamma are related to infectious complications rather than acute GvHD. Serum concentrations of IL-2 in most patients are undetectable.

Depletion of Foxp3+ regulatory T cells but not the absence of CD19+IL-10+ regulatory B cells hinders tumor growth in a para-orthotopic neuroblastoma mouse model.

Adaptive immune cells with regulatory function reportedly mediate immune escape in a variety of tumors. Little is known regarding the relevance of the most prominent regulatory cell populations, namely Foxp3+ T regulatory cells (Tregs) and CD19+IL-10+ B regulatory cells (Bregs), for neuroblastoma (NB) survival. After establishing a novel immunocompetent syngeneic NB mouse model where orthotopic tumors can be generated after intrarenal injection of NB975A cells, we studied the importance of Tregs and Bregs in Foxp3-DTR mice whose Tregs can be depleted by diphtheria toxin (DT) application as well as in CD19-specific IL-10 deficient mice that lack IL-10+ Bregs (CD19cre+/- × IL-10fl/fl mice). We observed Foxp3 Treg cells in tumors from wild type mice. On the contrary, Bregs or B cells were scarce. Specific depletion of Tregs in Foxp3-DTR mice resulted in an 85% reduction of tumor volume and weight compared to DT-treated wild type mice and untreated Foxp3-DTR mice. In contrast, NB tumor growth was not affected in CD19-specific IL-10 deficient mice. Similarly, mice lacking mature B cells (μMT mice) and CD19 deficient mice (CD19cre mice) showed no change in growth pattern of NB tumors. In Treg-depleted mice, reduced tumor growth was associated with an increased concentration of IFN-gamma, TNF-alpha, IL-4, IL-6, and IL-10 in isolated splenocytes. In summary, transient ablation of Tregs but not absence of Bregs hindered the growth of NB, strongly suggesting the therapeutic potential of targeting Tregs for this aggressive childhood tumor.

Assessment of the RANTES Level Correlation and Selected Inflammatory and Pro-Angiogenic Molecules Evaluation of Their Influence on CRC Clinical Features: A Preliminary Observational Study.

Background and Objectives: Assessment of RANTES level and concentrations of inflammatory cytokines: programmed death ligand 1 (PD-L1), interferon gamma IFN-γ, tumor necrosis factor alpha (TNF-α), transforming growht factor β (TGF-β) (and angiogenesis factors: vascular endothelial growth factor A (VEGF-A) and vascular endothelial growth factor C (VEGF C) in tumor and margin tissues of colorectal cancer (CRC,) and evaluation of RANTES influence on histopathological parameters (microvessel density (MVD), budding, tumor-infiltrating lymphocytes (TILs)), in relation to patients’ clinical features. Materials and Methods: The study used 49 samples of tumor and margin tissues derived from CRC patients. To determinate the concentration of RANTES, PD-L1, IFN-γ, TNF-α, TGF-β, VEGF-A, and VEGF-C, we used the commercially available enzyme-linked immunosorbent assay kit. Additionally, RANTES and PD-L1 expression was assessed with the use of IHC staining in both tumor cells and TILS in randomly selected cases. MVD was assessed on CD34-stained specimens. The MVD and budding were assessed using a light microscope. Results: We found significantly higher levels of RANTES, PD-L1, IFN-γ, TNF-α, TGF-β, VEGF-A, and VEGF-C in the tumor in comparison with the margin. The RANTES tumor levels correlated significantly with those of PD-L1, TNF-α, TGF-β, VEGF-A, and VEGF-C. The RANTES margin levels were significantly associated with the margin levels of all proteins investigated—PD-L1, IFN-γ, TNF-α, TGF-β, VEGF-A, and VEGF-C. Additionally, we observed RANTES- and PD-L1-positive immunostaining in TILs. In a group of 24 specimens, 6 different CRC tumors were positive for RANTES and PD-L1 immunostaining. The IFN-gamma concentration in both tumor and margin and TGF-β in tumor correlated with TILs. TILs were negatively associated with the patients’ disease stage and N parameter. Conclusions: RANTES activity might be associated with angiogenesis, lymphogenesis, and immune escape in CRC. RANTES is an important chemokine that is a part of the chemokine–cytokine network involved in the modulation of TME composition in CRC. Further research may verify which processes are responsible for the associations observed in the study.

Stromal Vascular Fraction and Mesenchymal Stem Cells from Human Adipose Tissue: A Comparison of Immune Modulation and Angiogenic Potential.

In recent years, both stromal vascular fraction (SVF) from adipose tissue and mesenchymal stem cells (MSC) from adipose tissues were extensively used in both preclinical and clinical treatment for various diseases. Some studies reported differences in treatment efficacy between SVFs and MSCs in animals as well as in humans. Therefore, this study is aimed to evaluate the immune modulation and angiogenic potential of SVFs and MSCs from the same SVF samples to support an explanation when SVFs or MSCs should be used. The adipose tissue samples from ten female donors with consent forms were collected. SVFs from these samples were isolated according to the published protocols. The existence of mesenchymal cells that positive with CD44, CD73, CD90, and CD105 and endothelial progenitor cells that positive with CD31 and CD34 was determined using flow cytometry. Three samples of SVFs with similar percentages of mesenchymal cell portion and endothelial progenitor cell portion were used to isolate MSCs. Obtained MSCs were confirmed as MSCs using the ISCT minimal criteria. To compare the immune modulation of SVF and MSCs, the mixed lymphocyte assay was used. The lymphocyte proliferation, as well as IFN-gamma and TNF-alpha concentrations, were determined. To compare the angiogenic potential, the angiogenesis in quail embryo assay was used. The angiogenesis efficacy was measured based on the vessel areas formed in the embryos after 7days. The results showed that all SVF samples contained the portions of mesenchymal cells and endothelial progenitor cells. MSCs from SVFs meet all minimal criteria of MSCs that suggested by ISCT. MSCs from SVFs efficiently suppressed the immune cell proliferation compared to the SVFs, especially at ratios of 1:4 (1 MSCs: 4 immune cells). MSCs also inhibited the IFN-gamma and TNF-alpha production more efficiently than SVFs (p<0.05). However, in quail embryo models, SVFs triggered the angiogenesis and neovessel formation better than MSCs with more significant vessel areas after 7days (p<0.05). This study suggested that SVFs and MSCs have different potentials for immune modulation and angiogenesis. SVFs help the angiogenesis better than MSCs, while MSCs displayed the more significant immune modulation. These results can guide the usage of SVFs or MSCs in disease treatment.

Antibody and cytokine levels in visceral leishmaniasis patients with varied parasitemia before, during, and after treatment in patients admitted to Arba Minch General Hospital, southern Ethiopia.

Visceral leishmaniasis is a disease caused by disseminated Leishmania donovani infection which affects almost half a million people annually. Most of the patients are reported from the Indian sub-continent, Eastern Africa and Brazil. In this study, we aimed to determine the levels of antibodies and cytokines in visceral leishmaniasis patients and to examine associations of parasitemia with the clinical states of patients. A prospective study was carried out, enrolling a total of 48 active VL patients who were evaluated before, during different time points and, three months after treatment. Serum cytokine concentrations, antibody levels, parasitemia, laboratory (hematologic and biochemical) measurements, and clinical parameters were assessed. Counts of WBC and platelets, and measurements of hemoglobin (Hb) increased during treatment (P ≤ 0.05). Elevated levels of circulating IL-10, IFN-γ, and TGF-β1 were measured before treatment. The observed increase in serum IL-10 remarkably declined within 7 days after the start of treatment. Anti-leishmanial antibody index (AI) was high in all VL patients irrespective of spleen aspirate parasite grade before treatment and at different times during treatment. However, a significant (P ≤ 0.05) decrease of AI was observed 120 days post-treatment. IL-2 serum levels were below the detection limit at all sampling points. The present results suggest that IL-10, IFN-γ, and TGF-β1 can be used as markers of active visceral leishmaniasis. In addition, measuring circulating cytokines concentrations, particularly IL-10, in combination with other clinical evaluations, could be used as criteria for the cure. The observation that a high serum concentration of IFN-gamma at baseline was associated with low parasitemia deserves further investigations.

Channeling macrophage polarization by rocaglates increases macrophage resistance to Mycobacterium tuberculosis

SummaryMacrophages contribute to host immunity and tissue homeostasis via alternative activation programs. M1-like macrophages control intracellular bacterial pathogens and tumor progression. In contrast, M2-like macrophages shape reparative microenvironments that can be conducive for pathogen survival or tumor growth. An imbalance of these macrophages phenotypes may perpetuate sites of chronic unresolved inflammation, such as infectious granulomas and solid tumors. We have found that plant-derived and synthetic rocaglates sensitize macrophages to low concentrations of the M1-inducing cytokine IFN-gamma and inhibit their responsiveness to IL-4, a prototypical activator of the M2-like phenotype. Treatment of primary macrophages with rocaglates enhanced phagosome-lysosome fusion and control of intracellular mycobacteria. Thus, rocaglates represent a novel class of immunomodulators that can direct macrophage polarization toward the M1-like phenotype in complex microenvironments associated with hypofunction of type 1 and/or hyperactivation of type 2 immunity, e.g., chronic bacterial infections, allergies, and, possibly, certain tumors.

Channeling Macrophage Polarization Via Selective Translation Inhibition by Rocaglates Increases Macrophage Resistance to Mycobacterium Tuberculosis

Macrophages contribute to host immunity and tissue homeostasis via alternative activation programs. M1-like macrophages control intracellular bacterial pathogens and tumor progression. In contrast, M2-like macrophages shape reparative microenvironments that can be conducive for pathogen survival or tumor growth. An imbalance of these macrophages phenotypes may perpetuate sites of chronic unresolved inflammation, such as infectious granulomas and solid tumors. We have found that plant-derived and synthetic rocaglates sensitize macrophages to low concentrations of the M1-inducing cytokine IFN-gamma and inhibit their responsiveness to IL-4, a prototypical activator of the M2-like phenotype. Treatement of primary macrophages with rocaglates increased their resilience to oxidative stress, stimulated autophagy and killing of intracellular mycobacteria. Thus, rocaglates represent a novel class of immunomodulators that can direct macrophage polarization towards the M1-like phenotype in complex microenvironments associated with hypofunction of type 1 and/or hyperactivation of type 2 immunity, e.g. chronic bacterial infections, allergies and, possibly, certain tumors.

The content of hypoxia-inducible factors and mediators of immunosuppression in the blood in diseases associated with hypoxia

The aim of the study was to identify general patterns and features of changes in the content of hypoxia-inducible factors-1 and -2 in association with an imbalance of cytokines (IL-10, IL-13, galectin-2 and -9, IFN-gamma) in the blood in diseases associated with hypoxia. Materials and methods. We examined 25 patients with coronary heart disease (CHD) with heart failure II-III according to NYHA, 16 patients with chronic obstructive pulmonary disease (COPD) without exacerbation, 16 patients with infiltrative pulmonary tuberculosis (TB) before anti-TB therapy, and 18 relatively healthy donors. Plasma concentrations of HIF-1alpha, HIF-2alpha, IL-10, IL-13, galectins-2 and -9, and IFN-gamma were determined by enzyme-linked immunosorbent assay (ELISA). Results. Positive outcomes of quantity determination of HIF-2alpha in the blood (24.00 ± 8.54 %, 75.00 ± 10.83%, 43.75 ± 12.40% of patients, respectively, against «zero» values in healthy donors) and also signs of immunosuppression at normal plasma concentrations of HIF-1alpha were determined in diseases associated with chronic hypoxia (in patients with CHD, COPD, TB). Immunological insufficiency in CHD and TB is caused by a deficiency of IFN-gamma and galectin-2 in association with an excess of galectin-9 (in patients with CHD 1.10 [0.52; 2.60] pg/ml p = 0.038) or IL-13 (in patients with TB 0.81 [0.79; 1.40] pg/ml, p = 0.043), and in patients with COPD it is caused by a surplus of galectin-9 and IL-13 (8.50 [3.96; 15.00] pg/ml, p = 0.001 and 2.62 [1.20; 7.58] pg/ml, p = 0.002, respectively) at normal concentrations of IFN-gamma and galectin-2. The content of IL-10 in the blood tends to increase in CHD and COPD. Conclusion. In patients with CHD, COPD and TB, chronic hypoxia is associated with immunosuppression mediated by an imbalance of IL-10, IL-13, IFN-gamma, galectins (2 and 9) in the blood and the secretion of HIF-2alpha, which has the property to stimulate the differentiation of M2-macrophages synthesizing anti-inflammatory cytokines.

Human defensins and Th-1 cytokines in hepatitis C viral infection.

Introductionactive or chronic exacerbated forms of hepatitis C virus (HCV) infection subsequently progress to liver disease and human defensins has been determined to have some level of anti-viral properties invitro whilst the expression of T helper-1 cytokines is known to promote complete recovery from acute HCV infection. The study sought to determine relationship between these immune responses.Methodsa cross sectional descriptive study design was employed. Hundred and thirty-two individuals were assessed were assessed for to anti-HCV, HCV RNA, serum levels of human alpha defensins 1 (HAD-1) and human beta defensins 1 (HBD-1). T helper 1 cytokines (IL-2, IFN gamma, TNF alpha) secreted in serum were also analyzed using commercial ELISA assay. The study was conducted in Kumasi, Obuasi and Daboya in Ghana.Resultsthe serum mean concentrations of HAD-1, HBD-1, IL-2, IFN gamma and TNF alpha showed no significant difference in concentrations among participants with chronic, spontaneously recovered or negative to HCV infection (p>0.05). Persons with hepatitis B co-infection were more likely to develop chronic HCV infection (p=0.039). HAD-1 and HBD-1 showed significant positive association with IL-2 (p=0.000) whilst only HAD-1 positively correlated with IL-2 (p<0.000).Conclusionthe immunological markers determined had no association with the status of HCV infection. HAD-1 increased with increasing levels of IL-2. These findings suggest that during HCV infection, inflammatory response through the production of cytokines by IL-2 cells may affect the release of HAD-1 and HBD-1.

SAT-454 METFORMIN REDUCES P-GP EXPRESSION AND POTENTIATES ANTI-INFLAMMATORY ACTION OF CORTICOSTEROIDS IN LUPUS NEPHRITIS PATIENTS

Metformin activates AMP kinase and alters transcription of multiple cytokines. P-glycoprotein expression is linked to drug resistance and has been shown to be associated with higher disease activity in lupus Nephritis. Metformin inhibits expression of P-glycoprotein (P-gp) in cancer cells. There is scarce data on effect of metformin on P-gp expression and its impact on cytokine secretion by PBMC in lupus. PBMC of nine lupus Nephritis patients (mean age 30 years, all females) were cultured using RPMA medium and then stimulated with PMA/ionomycin, with or without increasing dose of metformin (0.01, 0.1,1,10 mMol/L) for 24 h. Cytokines IL-1beta, IL-6, IFN-gamma and IL-10 were analysed by ELISA in culture supernatant. Cell viability with metformin was assessed by MTT assay. P-gp expression was measured by flow-cytometry. In another experiment, PBMCs from four patients were cultured with metformin 0.1mMol, prednisolone 0.3micMol and metformin + prednisolone and the concentrations of IL-1, IFN gamma and IL-10 were measured in culture supernatant. In MTT assay, viability of cells was maintained across all concentrations of metformin used. Metformin decreased expression of P-gp in PBMC in dose dependent manner (p = 0.003). In the PBMC cultures with PMA and metformin, there was dose dependent decrease in concentration of IL-1 beta (p = 0.001), IL-6 (p = 0.007) and IFN gamma (p < 0.001) and increase in that of IL-10 (p = 0.014) and TGF beta (p = 0.005) at lowest concentration of metformin. PBMC culture with combination of metformin and prednisolone had lower concentration of IL-1, IFN-gamma and higher concentration of IL-10 than with prednisolone or metformin alone. Merformin reduces P glycoprotein expression, decreases pro-inflammatory cytokines, increases anti-inflammatory cytokines and potentiates effect of corticosteroids.

Оптимізація лікування вірусіндукованих загострень бронхіальної астми у дітей із використанням лізатів молочнокислих бактерій Lactobacillus rhamnosus V

Objective. To study the effectiveness of lactic acid lactic acid bacteria Lactobacillus rhamnosus V in children with virus-induced exacerbations of bronchial asthma. Materials and methods. Were examined 35 patients aged 3 to 16 years old with exacerbation of bronchial asthma in the background of acute respiratory infections. Twenty five children of the main group were in the complex therapy of virus induced asthma exacerbation and were treated by lysate of the Lactic acid bacteria Lactobacillus rhamnosus V ( Del-Immune V ® ) at a dose of 25 mg 1 capsule 2 times a day 30 minutes before meals for 20 days. Ten patients were in the comparison group and obtained standard therapy. General clinical, immunological, allergic and statistical studies were used. Results. Use of Del-Immune V ® in the complex treatment of children with virus-nduced exacerbation of asthma, has led to a significant decrease in the frequency and severity of bronchial obstruction syndrome, which increases the effectiveness of treatment. In the main group, there was a significant increase in the concentration of IFN-gamma and a decrease in the concentration of cysteine leukotrienes in serum, and an increase in the concentration of a saliva IgA in the dynamics of treatment compared with the children in the comparison group. Conclusions. Observed integrated therapeutic effect of Del-Immune V ® use, no side effects and allergic reactions, justifying the appropriateness of Del-Immune V ® inclusion in the complex therapy of virus-induced asthma exacerbations.

Overexpression of Tear Inflammatory Cytokines as Additional Finding in Keratoconus Patients and Their First Degree Family Members.

Keratoconus is a progressive corneal ectasia that may lead to severe visual impairment due to the irregular astigmatism caused by corneal thinning. In addition to its association with atopy, eye rubbing, or genetic component, late reports suggest the involvement of inflammation in the pathogenesis of the disease. Our aim was to determine the concentration of IL-4, IL-6, IL-10, RANTES, IFN gamma, and TNF alpha in the tear film of patients with keratoconus and their first degree family members. We analyzed forty-eight participants in an observational cross-sectional study. The diagnosis of keratoconus had to be confirmed in addition to a minimum of 47 D corneal refractive power by corneal topography readings provided by a Placido-based topography system and analysis of the pattern: irregular astigmatism with an asymmetric “bow-tie.” As for the other groups, the most important diagnosis criteria were a normal topographic pattern with a regular astigmatism. 17 keratoconus patients, 16 relatives, and 15 controls were recruited after clinical assessment as part of the research. The cytokine's mean values were similar in the keratoconus group and the relatives' samples but significantly higher compared to the controls. Important differences were found in IL-4 levels between keratoconus patients and relatives and between relatives and controls (mean difference of 302.42, p < 0.0016 and 219.16, p < 0.033, Tukey's HSD procedure). In the keratoconus group, using the CORR procedure, we found statistically strong correlations of IL-6 lacrimal concentrations with the disease stage (r = 0.56, p < 0.01), keratometry (r = 0.55, p < 0.02), pachymetry (r = −0.64, p < 0.048), and corneal hysteresis (r = −0.53, p < 0.02). Cytokine overexpression may be relevant for the inflammatory etiology of keratoconus. In conclusion, in the case of some first degree family members, the elevated tear biomarkers may represent a supplementary risk factor.

Increased IL-6 Levels Correlate with Severity of Lung Damage and Early Death in Tanzanian Patients with Pulmonary Tuberculosis

Background: There is an urgent need for clinically relevant host biomarkers of protection, disease activity, response to treatment and predicting long term tissue damage for patients with pulmonary tuberculosis (TB). We studied cytokine biomarkers in a longitudinal cohort of patients with pulmonary TB in order to predict pulmonary tissue damage or death. Methods: Two hundred and thirtyfour (234) Tanzanian adult patients (96 HIV-positive) with newly diagnosed pulmonary TB and seven healthy controls were enrolled. Serum concentrations of IFN-gamma, TNF-α, IL-2, IL-6, IL-10. IL-17A and IL-21 were determined using enzyme-linked immunosorbent assay (ELISA) kits on day 1, two and five months after anti-TB treatment initiation. Postero-anterior chest X-rays (CXR) were scored for lung injury. Correlations between cytokine concentrations, mortality and lung injury were determined using multivariate logistic regression. Findings: Patients with TB had significantly higher median levels of IL-6 (3863pg/ml) at diagnosis compared to healthy controls (11 pg/ml). Increased IL-6 levels significantly correlated with survival (RR = 0.95; 95% CI 0.91 - 0.98; P < 0.01) and were associated with severe lung injury (RR 1.03; 95% 1.01 - 1.05; P = 0.02). Among patients with severe lung injury at diagnosis, every 100pg/ml increase in baseline IL-6 levels was accompanied with 6% increased risk for residual lung damage post-TB treatment (RR = 1.06; 95%CI 1.01 - 1.13). Anti-TB therapy was associated with reduction in serum cytokine levels except for IL-6, which did not decrease with treatment (P = 0.003). Interpretation: Increased IL-6 levels correlate with severity of lung damage in patients with pulmonary TB regardless of HIV status. IL-6 may serve as a target for host-directed therapy in patients with pulmonary TB. Funding: This study was supported by grants from the Swedish International Development Cooperation Agency (SIDA); Vinnova and Swedish Heart and Lung Foundation (Hjartlungfonden) to MM. AZ and MM acknowledge support from the European and Developing Countries Clinical Trials Partnership (EDCTP). Declaration of Interest: The authors declare no conflicts of interest Ethical Approval: Ethical clearance was granted by the Muhimbili University of Health and Allied Health sciences (MUHAS) Senate Research and Publications committee (Ref. No. RP/AEC/VOL.XII dated November 23rd2011). Written informed consent was obtained from all study participants prior to inclusion.

Differences in Inflammatory Cytokine Levels between Patients with Varying Severity of Chronic Venous Insufficiency

Objective: Several studies have found that protein concentrations of inflammatory cytokines are significantly increased in tissue and serum from patients with chronic venous insufficiency (CVI) relative to that of healthy controls. We sought to determine whether inflammatory protein concentrations differ between patients with moderate clinical disease, classified as clinical, etiology, anatomy, pathophysiology (CEAP) Classes 2 and 3, and more severe clinical disease, classified as CEAP Class 4. Methods: Twenty patients with abnormal venous function were included in the study. Blood from a competent leg vein and from an incompetent superficial vein was collected in addition to incompetent vein tissue extracted through a phlebectomy procedure. Cytokine levels of venous tissue lysate and serum samples were determined using a multiplex assay. Results: Thirteen patients (65%) were classified as clinical CEAP Class 2 or 3, with seven patients (35%) falling into the more severe Class 4 category. Twenty-seven cytokines were measured. Serum isolated from normal veins had significantly higher levels of IFN-gamma in patients with Classes 2 and 3 than Class 4 disease (95.17 pg/mL vs. 71.97 pg/mL; p=0.036). In serum from incompetent veins, IFN-gamma concentrations averaged 95.47 pg/mL in Class 2 and 3 patients and 76.97 pg/mL in Class 4 patients (p=0.048). Eotaxin levels from diseased vein tissue averaged 3.37 pg/mL in Classes 2 and 3 patients, and 1.57 pg/mL in Class 4 patients (p=0.037). IP-10 levels in diseased vein tissue was also significantly less in Class 4 patients at 74.20 pg/mL in Class 2 and 3 patients versus 31.06 pg/mL in Class 4 patients (p=0.004). Conclusion: Despite several studies documenting increased inflammatory cytokines in patients with CVI, our study shows that those patients with more severe disease have significantly lower levels of several inflammatory cytokines. Thus, particular inflammatory cytokines may function in a reparative capacity following tissue injury or as a control mechanism to inhibit further tissue destruction.

Comparison of interferon and bovine herpesvirus-1-specific IgA levels in nasal secretions of dairy cattle administered an intranasal modified live viral vaccine prior to calving or on the day of calving

Thirty-two Holstein cows were allocated to receive intranasal vaccination with modified live bovine herpesvirus-1 (BHV-1), bovine respiratory syncytial virus (BRSV) and parainfluenza type 3 virus (PI3V) vaccine either two weeks prior to their projected calving date, or within 24h after calving. Nasal secretions were collected twice at a 12-h interval on the day prior to vaccination (day 0) and at 2, 4, 7, 10 and 14days post vaccination to measure interferon (IFN) alpha, IFN-beta, IFN-gamma, and BHV-1-specific IgA by ELISA. Serum neutralizing antibody titers to BHV-1 and BRSV were measured on days 0, 7, and 14.There was a significant treatment effect (p<0.0004) and interaction (p<0.05) on nasal BHV-1 IgA levels, with higher IgA levels in cows vaccinated within 24h after calving. There was a significant treatment effect on nasal IFN-gamma concentration (p<0.05) and on nasal total IFN concentration (p<0.05), with higher IFN-gamma and total IFN concentrations seen in cows vaccinated within 24h after calving. There was no significant treatment or interaction effect on nasal IFN-alpha or IFN-beta concentrations, or on serum neutralizing titers to BRSV.In spite of prior viral vaccination during the previous lactation, cows vaccinated on the day of calving responded to an intranasal viral vaccination with increased concentrations of IFN-gamma and increased titers of IgA following vaccination which was significantly higher than cows vaccinated precalving. This study is the first to examine respiratory mucosal responses in immunologically mature dairy cattle vaccinated intranasally before and after calving.