Evaluation of the relationship between IL-12, IL-13 and TNF-α gene polymorphisms with the susceptibility to brucellosis: a case control study.

BACKGROUND: The cytokine gene polymorphism is important for the genetic susceptibility of infectious diseases. The aim of the present study was to investigate the relationship between TNF-α, IL-12, and IL-13 gene polymorphisms and predisposition to brucellosis. METHODS: In this study, 107 patients with brucellosis and 107 healthy individuals were evaluated. The SNPs of TNF-α)- 238 G/A) and IL-12 (+ 1188 A/C) were done by amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) and IL-13 genotyping at positions - 1512 (A/C) and - 1112 (C/T) were analysis by restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR) methods. IL-12, IL-13 and TNF-α serum levels were measured by a sandwich enzyme-linked immunosorbent assay (ELISA). RESULTS: IL-13 (-1512A/C) was associated with Brucellosis risk in dominant model (OR (95% CI) = 2.17 (1.02-4.62)), P-value = 0.041. However, there was no difference in allele and genotype frequencies of TNF-α)- 238 G/A), IL-12 (+ 1188 A/C) and IL-13 [- 1512 (A/C) and - 1112 (C/T)] between patients and controls. Serum levels of IL-12 and TNF-α were significantly more frequent in the patients than in the control groups. CONCLUSIONS: The IL-13 gene polymorphism can be used as a biomarker for detecting susceptibility to Brucella disease.

Effect of 50-Hz Magnetic Fields on Serum IL-1ß and IL-23 and Expression of BLIMP-1, XBP-1, and IRF-4.

Investigations demonstrated that magnetic fields (MFs) change cytokine production and expression of some immune system genes. This alteration can affect the immune system function and may lead to some diseases. Therefore, this study investigated two important inflammatory cytokines, i.e., IL-1ß and IL-23 at two phases of pre- and post-immunization of the immune system. In addition, the expressions of three important genes in the humoral immunity, i.e., B lymphocyte-induced maturation protein-1 (BLIMP-1), X-box-binding protein-1 (XBP-1), and interferon regulatory factor-4 (IRF-4) were evaluated at post-immunization phase. Eighty adult male rats were divided into four experimental groups and a control. The experimental groups were exposed to 50 -Hz MFs with magnetic flux densities of 1, 100, 500, and 2000 µT, 2 h/day for 2 months. The animals were injected by human serum albumin (100 µg/rat) on days 31, 44, and 58 of exposure. The cytokine levels in serum were measured with enzyme-linked immunosorbent assay (ELISA), and the expression of genes was evaluated with reverse transcription quantitative polymerase chain reaction (RT-qPCR). Serum IL-1ß was decreased at pre-immunization phase after exposure to 1 and 100 µT of 50-Hz MFs. In contrast, serum IL-23 was increased at post-immunization phase in 100 µT group. No change was observed in serum IL-1ß and IL-23 in each group at pre-immunization phase compared with post-immunization. Furthermore, exposure to 100 µT downregulated expression of BLIMP-1, XBP-1, and IRF-4. In conclusion, exposure to 50-Hz MFs may decrease inflammation at short time and increase it at longer time exposures. In addition, 50-Hz MF exposure may decrease the humoral immune responses. It seems that 50-Hz MFs cause more alteration in immune system function at lower densities (100 µT).

The increased T helper cells proliferation and inflammatory responses in patients with type 2 diabetes mellitus is suppressed by sitagliptin and vitamin D3 in vitro.

OBJECTIVE: The probably effects of sitagliptin and vitamin D3 (VitD3) on proliferation capacity and cytokines production were investigated in type 2 diabetes mellitus (T2DM) in vitro. MATERIALS AND METHODS: Peripheral blood mononuclear cells (PBMCs) were isolated from 35 patients with T2DM and 26 healthy controls (HCs). CFSE-labeled PBMCs stimulated with phytohamagglutinin (PHA, 5 µg/mL) in the presence/absence of sitagliptin (200 mg/mL) with/without VitD3 (10-8 M) for 4 days. The proliferation of CD4+ T helper cells and non-CD4+ cells was analyzed using flow cytometry. The supernatant levels of IFN-γ, IL-17, IL-4, TGB-ß and IL-37 were detected using ELISA. RESULTS: The proliferation of CD4+ T cells in response to PHA was higher in T2DM patients compared with HCs. The production of IFN-γ and IL-17 in PHA-stimulated cultures was higher, and the levels of IL-4 and IL-37 were lower in T2DM patients compared to HCs. The addition of sitagliptin or VitD3 to the cultures decreased the CD4+ T cells and non-CD4+ cells proliferation in patients and HCs. Sitagliptin with VitD3 was more effective in suppression of proliferation, decreasing of IL-17 and enhancing of IL-37 production. CONCLUSION: Sitagliptin plus VitD3 effectively reduces the proliferative T cells response and modulates pro-inflammatory/anti-inflammatory cytokines production.

Anti-Inflammatory Effect of Combined Sitagliptin and Vitamin D3 on Cytokines Profile in Patients with Type 2 Diabetes Mellitus.

Studies indicated that imbalance of proinflammatory and anti-inflammatory cytokines may contribute to development of type 2 diabetes mellitus (T2DM). We hypothesized that sitagliptin and VitD3 may exert more anti-inflammatory effects on the regulation of cytokine balance in T2DM. Nonnephropathic and nephropathic T2DM patients were divided into the subgroups, based on treatments. The effect of 8 months sitagliptin, alone or together with 2 months of VitD3, on serum IFN-γ, IL-4, IL-17, IL-6, IL-21, TGF-ß, and IL-37 levels was determined using enzyme-linked immunosorbent assay. Increased levels of interferon (IFN)-γ and IL-17 in untreated (without sitagliptin and VitD3) nephropathic and nonnephropathic patients and decreased levels of IL-37 in untreated nephropathic patients were observed compared with healthy controls. Treatment with sitagliptin plus VitD3 reduced the levels of IFN-γ and IL-17 in both nonnephropathic and nephropathic patients compared with untreated patients. The level of IL-37 was enhanced in patients treated with sitagliptin or sitagliptin plus VitD3, compared with untreated patients. Sitagliptin plus VitD3 treatment increased the levels of IL-4 in nonnephropathic patients. These findings indicated that the sitagliptin plus VitD3 was more effective to reduce the increased proinflammatory IFN-γ and IL-17 cytokines in T2DM patients.

Effects of sitagliptin and vitamin D3 on T helper cell transcription factors and cytokine production in clinical subgroups of type 2 diabetes mellitus: highlights upregulation of FOXP3 and IL-37.

Objective: Gene expression level of T helper cell transcription factors and cytokines production in type-2 diabetes mellitus (T2DM) patients treated with mono- or combined sitagliptin and vitamin D3 (VitD3) were evaluated. Methods: Fifty-four nephropathic and 57 non-nephropathic T2DM patients were divided into the subgroups based on their treatment with/without sitagliptin and VitD3. The expression of T-bet, RORγt, BCL6, and FOXP3 was evaluated using real-time PCR. The levels of IFN-γ, IL-6, IL-17, IL-21, TGF-ß, and IL-37 were assessed in PBMC supernatants using ELISA. Results: The production of IFN-γ and IL-17 was increased in untreated (without sitagliptin and VitD3) nephropathic and non-nephropathic T2DM patients compared with healthy controls, whereas FOXP3 expression was decreased. Treatment with sitagliptin alone or in combination with VitD3 reduced the production of IFN-γ in the patients. Production of IL-17 and IL-21 and the expression of RORγt and BCL6 was diminished in patients treated with combined sitagliptin and VitD3, whereas the production of IL-37 and FOXP3 expression were increased in the patients treated with sitagliptin or sitagliptin plus VitD3. Conclusion: These data demonstrate that sitagliptin in combination with VitD3 may accelerate the process of T2DM treatment by exerting synergic anti-inflammatory effects on immune system through upregulation of FOXP3 and IL-37, and downregulation of RORγt and BCL6 as well as IFN-γ, IL-17 and IL-21 production. Combined sitagliptin and VitD3 can be safely utilized to modulate the inflammatory conditions of T2DM.

Decreased regulatory function of CD4+CD25+CD45RA+ T cells and impaired IL-2 signalling pathway in patients with type 2 diabetes mellitus.

In this study, the frequency and function of CD4+CD25+CD45RA+ regulatory T cells (Treg) and intracellular IL-2 signalling molecules in patients with type 2 diabetes mellitus (T2DM) were investigated. Tregs and responder T cells (Tresp, CD4+CD25- T cells) were sorted and suppression assays were performed using flow cytometry. Phosphorylation of signal transducer and activator of transcription-5 (pSTAT5) were assessed using flow cytometry. Gene expression of FOXP3 was performed with the SYBR green Real Time PCR method. Production of IL-2 from cultured cells was assessed using ELISA. We observed a functional defect of CD4+CD25+CD45RA+ Tregs in T2DM patients with higher proliferation of Tresp cells, in response to anti-CD3 and anti CD28 stimulation in the presence of Tregs in vitro. The results showed that the proliferation of Tresps in the absence of Treg cells was higher in T2DM patients than in healthy controls. Decreased FOXP3 mRNA expression and pSTAT5 were observed within the Tregs of the patients, whereas the level of secreted IL-2 from PBMCs culture was not statically different between T2DM patients and healthy individuals. Changes in intracellular IL-2 pathways and FOXP3 gene expression may contribute to the defect of Tregs in T2DM patients. These findings indicating that the purified CD4+CD25+CD45RA+ Treg cells have reduced functional capacity together with impaired IL-2 pathway in T2DM, and the Tregs could be used for a potential novel therapeutic target.

Extremely Low Frequency Electromagnetic Fields Decrease Serum Levels of Interleukin-17, Transforming Growth Factor-ß and Downregulate Foxp3 Expression in the Spleen.

The study aimed to determine effect of extremely low frequency (50 Hz) electromagnetic fields (ELF-EMFs) exposure on serum levels of interleukin-17 (IL-17) and transforming growth factor-ß (TGF-ß) as signature cytokines of Th17 and regulatory T (Treg) cells, respectively. Retinoid-related orphan receptor γT and transcription factor forkhead box P3 (Foxp3) expression levels as lineage defining of Th17 and Treg cells were also assessed in the spleen and thymus. Eighty male rats were separated into 4 exposed groups (1, 100, 500, and 2,000 µT magnetic flux intensities) and a control. All rats were immunized by human serum albumin after 1 month of the exposure and the experiment was continued in the same manner for 1 month more. The results demonstrated that the weight of thymuses was significantly declined at intensity of 2,000 µT. At the preimmunization phase, the serum levels of IL-17 and TGF-ß were significantly decreased at intensities of 1 and 100 µT. The expression of Foxp3 was also downregulated at intensities of 1 and 100 µT. In conclusion, low intensities of ELF-EMF may reduce the serum levels of IL-17 and TGF-ß and downregulate the expression of Foxp3 in spleen.

Effect of Cinnamon and Turmeric Aqueous Extracts on Serum Interleukin-17F Level of High Fructose-Fed Rats.

BACKGROUND: Studies have indicated that extraweight and obesity induce chronic inflammation, which can lead to other diseases such as cancers. OBJECTIVE: To evaluate the effects of two weight-lowering and anti-inflammatory agents including cinnamon, and turmeric, on serum levels of interleukin-17 (IL-17) as a pro-inflammatory cytokine. METHODS: In this study, 64 rats were designated in eight groups. The control group received normal diet. The other groups were fed with normal diet plus high cinnamon (3 mg/ml), high turmeric (3 mg/ml), high-fructose solution (30%), fructose solution with low (0.15 mg/ml) and high doses (3 mg/ml) of cinnamon and turmeric three times per week. The serum level of IL-17F was measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: High fructose consumption led to an increase in the weight and serum level of IL-17. While, feeding with cinnamon and turmeric caused to decline weight but, surprisingly increased IL-17F levels. CONCLUSION: Although, some studies have showed that cinnamon and turmeric supplementation decreased IL-17F under the standard diet, in the presence of high fructose diet and extraweight their effects were reversed and caused an increase in serum level of IL-17F.

Vitamin D3 inhibits the proliferation of T helper cells, downregulate CD4+ T cell cytokines and upregulate inhibitory markers.

1 α, 25-dihydroxyvitamin D3 (VitD3) has been suggested to have strong modulatory properties in the immune system. Researchers in the present study primarily aimed to understand the effect of VitD3 on human CD4+ T cell proliferation in antigen presenting cells (APCs) free condition in vitro. The effect of VitD3 on intracellular cytokine responses trend to Th1, Th2, Th17 and Th22 was evaluated using the flow cytometry. Moreover the effect of VitD3 on the expression of inhibitory markers such as PD1, PD-L1, and CTLA4 which are induced upon polyclonal T cell receptor (TCR) activation on CD4+ T cells, was assessed. We observed that the stimulation of CD4+ T cells with VitD3, suppressed proliferation capacity, enhanced the expression of PD1, PD-L1 and CTLA4 inhibitory markers on CD4+ T cells, and diminished the percentage of pro-inflammatory cytokines including, IFN-γ, IL-17, and IL-22 except IL-4 in CD4+ T cells. The data suggested a potential insight into the consideration of VitD3 in the prevention/control of pro-inflammatory immune response/autoimmune disorders.

Moderate Exercise Enhances the Production of Interferon-γ and Interleukin-12 in Peripheral Blood Mononuclear Cells.

The purpose of this study was to explore the effect of two months moderate exercise on levels of IFN-γ, IL-12, IL-6 and IL-4 in serum and supernatants of in vitro mitogen-activated (PHA for 48 h) whole blood (WB) and peripheral blood mononuclear cells (PBMCs). Sixteen healthy males participated in running program (30 min/day, 5 days/week). Blood samples were collected in three stages; 24 h before to start exercise, 48 h and two months after the last session of the exercise. The samples were analyzed for the cytokines by ELISA. The levels of IFN-γ and IL-12 were increased significantly in activated PBMCs culture after exercise and were back to normal level after two months rest. A significant elevation of IFN-γ/IL-4 ratio was observed in activated PBMCs culture by acting possibly on IFN-γ. The results suggest that short moderate intensity exercise enhances Th1 immune inflammatory and anti-allergic conditions in response to mitogen.

Genetic heterogeneity within the HLA region in three distinct clinical subgroups of myasthenia gravis.

This study aims to investigate genetic susceptibility to early-onset and late-onset anti-acetylcholine receptor antibody positive myasthenia gravis (EOMG and LOMG) and anti-muscle specific kinase antibody positive MG (MuSK-MG) at genome-wide level in a single population. Using a custom-designed array and imputing additional variants and the classical HLA alleles in 398 patients, we detected distinct associations. In EOMG, rs113519545 in the HLA class I region (OR=5.71 [3.77-8.66], P=2.24×10(-16)), HLA-B*08:01 (OR=7.04 [3.95-12.52], P=3.34×10(-11)) and HLA-C*07:01 (OR=2.74 [1.97-3.81], P=2.07(-9)), in LOMG, rs111256513 in the HLA class II region (OR=2.22 [1.59-3.09], P=2.48×10(-6)) and in MuSK-MG, an intronic variant within HLA-DQB1 (rs68081734, OR=5.86, P=2.25×10(-14)) and HLA-DQB1*05:02 (OR=8.56, P=6.88×10(-13)) revealed the most significant associations for genome-wide significance. Differential genetic susceptibility within the HLA to EOMG, LOMG and MuSK-MG has been established in a population from Turkey.

The role of T regulatory cells in immunopathogenesis of myasthenia gravis: implications for therapeutics.

T regulatory cells (Tregs) are crucial for the development of self-tolerance and are the major focus in many studies interpreting the pathogenesis of myasthenia gravis (MG), an autoimmune-based disease. In normal conditions, Tregs regulate the immune responses, while impaired regulatory function of these cells can lead to autoimmunity. Recent studies have confirmed that the thymic and peripheral blood CD4(+)CD25(+) Tregs of MG are defective in functions and/or in numbers, which are associated with disease severity; approaches to correct the defects of these Tregs may be promising in the treatment of MG. This review discusses recent studies on characteristics, quantitative and qualitative changes of Tregs and possible mechanisms that are involved in the impairment of these cells in MG pathogenesis. In addition, new approaches inducing Treg generation that are currently being investigated as therapies for MG, will be discussed as well as proposed approaches for future therapies.

Regulatory function of CD4+CD25++ T cells in patients with myasthenia gravis is associated with phenotypic changes and STAT5 signaling: 1,25-Dihydroxyvitamin D3 modulates the suppressor activity.

Regulatory T cells were investigated in early-onset (EO) and late-onset (LO) myasthenia gravis patients with anti-acetylcholine receptor antibody (AChR-MG). Alterations in PD-1 and PD-L1 on CD4(+)CD25(++) (Treg) and responder T cells (Tresp, CD4(+)CD25(-)) were observed in LOMG patients. GITR was decreased on CD4(+)CD25(++) of all patients. Decrease of FOXP3 was associated with lower phosphorylation of STAT5.1,25-dihydroxyvitamin D3 (VitD3) increased suppression in co-culture with a stronger effect in patients by acting possibly both on cell groups. Changes in surface molecules and intracellular pathways contribute to the defects of Treg in non-thymomatous AChR-MG and VitD3 can have modulatory effects.

Association of HLA-DRB1∗14, -DRB1∗16 and -DQB1∗05 with MuSK-myasthenia gravis in patients from Turkey.

Susceptibility to myasthenia gravis (MG) has been demonstrated with several HLA in different disease subgroups. HLA-DR14, -DR16 and -DQ5 were reported as predisposing factors in muscle-specific kinase antibody positive MG (MuSK-MG). These markers were evaluated in MG subgroups from Turkey. Among 164 generalized MG patients, 116 had antibodies against anti-acetylcholine receptor (AChR-MG) and 48 had MuSK-MG. Only HLA-DRB1 and DQB1 allele groups reported to be associated with MuSK-MG were compared with 250 healthy controls (HC). Highly significant associations of both DRB1(∗)16 and DRB1(∗)14 were found with MuSK-MG compared to HC (p = 1.9 × 10(-5), OR: 4.95 and p = 0.0028, OR: 3.1). On the contrary, HLA-DRB1(∗)03 was less frequent in MuSK-MG (p = 0.006, OR: 0.09). DQB1(∗)05 was also associated with MuSK-MG (p = 2.5 × 10(-6) OR: 4.8). This study provides a replication of the highly significant associations of both HLA-DRB1(∗)16,-DRB1(∗)14 and -DQB1(∗)05 with MuSK-MG. Moreover, HLA-DRB1(∗)03 appears to have a distinguishing role for this disease subgroup compared to early-onset and AChR-MG.