Search results for: Human IL-13 ELISA Kit





The role of microglial P2X7: modulation of cell death and cytokine release.
ATP-gated P2X7 is a non-selective cation channel, which participates in a wide range of cellular functions as well as pathophysiological processes including neuropathic pain, immune response, and neuroinflammation. Despite its abundant expression in microglia, the role of P2X7 in neuroinflammation still remains unclear.
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Interleukin-13 promoter gene polymorphism -1112 C/T is associated with atopic dermatitis in Polish patients.
Interleukin 13 (IL-13) is one of the major cytokines involved in the IgE synthesis in patients with atopic dermatitis (AD). IL-13 gene has been mapped on chromosome 5q31-33 region associated with atopic conditions, where several polymorphisms are described. The aims of the study were to establish the frequency of the IL-13 gene polymorphism and its relation to serum IgE and IL-13 levels and SCORAD. In 180 AD patients and 167 healthy volunteers, the -1112 C/T IL-13 gene polymorphism was genotyped using allele-specific PCR method. Serum total IgE (tIgE) level was measured by Uni-CAP 100 and IL-13 level using ELISA standard kit. The -1112T allele frequency was significantly higher in AD patients compared to controls (P=0.00001). The TT and CT genotypes were correlated with increased serum tIgE concentration (P=0.0004). The TT genotype was associated with severe, CT genotype with moderate and CC genotype with mild course of AD (P=0.0008). Both CT and TT genotypes predominated in cases of AD with concomitant asthma, while CC genotype was not found in any case in this group (P=0.03). The mean levels of serum IL-13 and tIgE were significantly higher in AD with concomitant asthma than in cases without asthma (IL-13 P=0.03 and tIgE P=0.0001). There was positive correlation between serum concentration of IL-13 (P=0.0005) and tIgE (P=0.0000001) and severity of AD as assessed by SCORAD index. Our results confirmed the significant role of -1112 C/T IL-13 gene polymorphism in the pathogenesis of AD.
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interleukin 17 receptor C
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A common variant associated with asthma, interleukin 13 R130Q, promotes the production of IgE.
Interleukin (IL)-13 plays an important role in the pathogenesis of asthma. A polymorphic variant of human IL-13 R130Q, results in substitution of an arginine with a glutamine was shown to be associated with asthma in Chinese Han nationality. We examined the functional consequences of this variant in vitro to investigate whether this variant enhanced functional activity compared with wild type IL-13. The wild-type and mutant IL-13 genes were amplified from the plasmid of pET22b-hIL-13 by PCR and site-directed mutagenesis PCR. Both the PCR product and the vector pET28a(+) were digested by the NdeI and BamHI. Then the PCR product was cloned in the prokaryotic expression vector of pET28a(+). The plasmids were constructed and transformed into E. coli BL21(DE3). The positive clones were selected, and tested by sequencing. Peripheral blood mononuclear cells (PBMCs) from healthy participants were isolated and cultured with increasing concentrations of recombinant WT IL-13 and IL-13 R130Q. IgE was detected with ELISA kit in the supernatants. Recombinant WT IL-13 and IL-13 R130Q were successfully expressed into the prokaryotic expression system and their biological activity was consistent with standard protein. Our results show that IL-13 R130Q is more active than WT IL-13 in inducing hydrocortisone-dependent IgE synthesis. There were statistical significances between them. IgE induction by physiologic concentrations was obviously increased. IL-13 R130Q has increased activity compared with wild type IL-13 in vitro. And IL-13 R130Q may be used for new target of asthma for diagnosis and therapy in the future.
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Plant chitinase III Ziz m 1 stimulates multiple cytokines, most predominantly interleukin-13, from peripheral blood mononuclear cells of latex-fruit allergic patients.
Indian jujube is a fruit abundantly cultivated in Taiwan. Its major allergen in latex-fruit syndrome is Ziz m 1 of the chitinase III family. The Ziz m 1 Pichia (rZiz m 1-P) has chitinase activity but not Ziz m 1 E. coli (rZiz m 1-E).
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IL-13 gene expression in patients with atopic dermatitis: relation to IgE level and to disease severity.
Atopic dermatitis (AD) is a chronic inflammatory skin disease frequently associated with an increased serum IgE level. T helper cells are thought to play an important role in the pathogenesis of AD. It is commonly believed that allergens activate Th2 cells, and it is likely that the cytokines produced by Th2 cells are crucial factors in the induction and maintenance of the disease. IL-13 is one of the cytokines that are produced by Th2 lymphocytes and, like IL-4, it can induce the production of IgE. In order to evaluate its role in the pathogenesis of AD, IL-13 mRNA expression was studied in peripheral blood of patients with different degrees of AD and compared with healthy subjects. Also, we correlated its level of expression with the level of serum IgE and with the severity of the disease. EDTA blood was obtained from 25 patients (divided into three groups ranged from mild to severe AD) and 12 normal subjects as a control group. We examined the blood sample for IL-13 mRNA expression using RNA extraction technique, RT-PCR, PCR amplification using primers specific for IL-13 and beta- actin (as internal control) this is followed by visualization of the expressed bands using gel electrophoresis and DNA marker. Serum IgE level was detected using an ELISA kit. Our results revealed that, IL-13 mRNA is significantly expressed in patients with AD as compared to normal control (P<0.001). IL-13 mRNA shows higher level of expression in severe AD group in comparison with both moderate and mild groups (P = 0.05). Serum levels of IgE showed highly significant increase in patients with AD as compared with the control group (p=0.019), its level is significantly higher in severe AD group versus moderate and mild AD groups (P=0.009 and 0.022, respectively). There is a highly significant positive correlation between serum levels of IgE and the levels of IL-13 mRNA expression in all AD groups (P=0.001). In conclusion, the high level of IL-18 mRNA expression in AD, and its correlation with serum level of IgE and with severity of disease indicates that IL-13 is involved in the pathogenesis of the disease and is an important in vivo IgE inducer.
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[Interleukin levels in umbilical cord blood: relationship with a family history of allergic disease].
The development of allergic pathology takes place as result of an alteration of the immunity and alteration of the corporal mechanism of protection, giving rise to an erroneous answer or exaggerated forehead to innocuous antigens, that it generates clinical symptoms with cutaneous, digestive or respiratory manifestations. The frequency and distribution of this process have undergone an increase from the 1970, which causes that a greater interest in the knowledge of the mechanisms exists that produce this clinic. The answer by immunoglobulin E is regulated by the answer of lymphocytes T-helper-1 represented by interleukin 2 and gamma-interferon that inhibits their production, and the answer of lymphocytes T-helper-2 formed by interleukin 4, interleukin 10 and interleukin 13 that stimulate the production of immunoglobulin E.
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Synergistic effect of SCF and TNF-alpha on the up-regulation of cell-surface expression of ICAM-1 on human leukemic mast cell line (HMC)-1 cells.
Intercellular adhesion molecule-1 (ICAM-1) has been shown to play crucial roles in mast cell interaction with other inflammatory cells and recruitment into the inflamed tissue. In the present study, human mast cell line-1 (HMC-1) was stimulated with different cytokines including stem cell factor (SCF), tumor necrosis factor alpha (TNF-alpha), interleukin (IL)-13, IL-18, and IL-25. Cell-surface expression of ICAM-1 was assessed by flow cytometry. To elucidate the intracellular signal transduction regulating the ICAM-1 expression, phosphorylated extracellular signal-regulated kinase (ERK), phosphorylated p38 mitogen-activated protein kinase (MAPK), and nuclear factor (NF)-kappaB translocation were assessed by enzyme-linked immunosorbent assay. Results showed that SCF, TNF-alpha, and IL-13 but not IL-18 and IL-25 could up-regulate the surface expression of ICAM-1 on HMC-1 cells. A synergistic effect of SCF and TNF-alpha on ICAM-1 expression was demonstrated. This synergistic effect was shown to be dose-dependently enhanced by SCF but not TNF-alpha. Results indicated that SCF activated ERK, and TNF-alpha activated the p38 MAPK and NF-kappaB pathway. Selective inhibitor of ERK, PD098059, and c-kit inhibitors, STI571 and PP1, suppressed the combined SCF and TNF-alpha-induced ICAM-1 expression. BAY117082 but not SB203580, which are the inhibitors of NF-kappaB and p38 MAPK, respectively, suppressed the TNF-alpha-induced ICAM-1 expression. Therefore, SCF and TNF-alpha acted through ERK and the NF-kappaB pathway to regulate the ICAM-1 expression and elicited the synergistic effect. In conclusion, our results provide insight for cross-talk between different signaling pathways that can help in understanding the fine control of adhesion molecule expression under the concerted effects of cytokines.
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Production of IL-13 by human lung mast cells in response to Fcepsilon receptor cross-linkage.
Cross-linkage of the high affinity Fcepsilon receptors (FcepsilonRI) on the surface of the mast cell by the allergen-IgE complex is a central event in the induction of allergic inflammatory reactions. However, the precise roles of human mast cells in the perpetuation of allergic inflammation is not well known. IL-13 plays an important role in the regulation of allergic inflammation, especially being involved in the induction of IgE synthesis.
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