mRNA Expression Of Cytokine Genes Research Articles

Effects of environmental salinity on the immune response of the coastal fish Scatophagus argus during bacterial infection

The coastal aquaculture is characterized with environmental salinity fluctuation, and the effects of salinity stress on the immunity of cultured fish are needed to be further explored. Scatophagus argus is an important species in the wild fisheries and aquaculture industry, it would be of great value to reveal the impact of salinity change on the immune response in this species. Understanding the effects of salinity stress on immune response can provide valuable insights into salinity management in the aquacultural process. The head kidney, which is an organ unique for teleost fish, functions not only as a central immune organ but also as a crucial role in the stress response during which the secretion of immunoregulatory molecules i.e. cytokines is facilitated. In the present study, Individuals of S. argus acclimated to 3 different salinities [0‰ (FW), 10‰ (BW), and 25‰ (SW)] were injected intraperitoneally with A. hydrophila, and then monitored throughout one week. The effects of environmental salinity on the immune response in S. argus stimulated by A. hydrophila infection were investigated. mRNA expression profiles of cytokine genes IL-1β, IL-6, IL-10 and TNF-α in different salinity groups was quite different. mRNA expression of cytokine genes in BW group and SW group rose more quickly and significantly higher than FW group (p < 0.05) at early stages (6–24 hpi) after bacterial injection, and before 96 hpi, the highest value of cytokine expression at each time point was recorded in SW group. Immune parameters such as lysozyme level, complement C3 activity and IgM content in BW and FW groups were lower than SW group at each time point from 24 to 144 hpi after bacterial injection. In addition, leukocyte profiles in the head kidney and blood were also investigated. Although hypoosmotic acclimation could temporarily stimulate monocyte and neutrophil proliferation, it was observed that the number of monocytes, neutrophils and lymphocytes of the head kidney and blood in SW group increased more quickly than BW and FW groups after bacterial infection. Our results indicate that hypoosmotic stress due to the decrease of environmental salinity has suppressive immunoregulatory effects on the immune response of S. argus.

Effects of Dietary Nano-Selenium Supplementation on Growth Performance, Antioxidative Status, and Immunity in Broiler Chickens under Thermoneutral and High Ambient Temperature Conditions.

The objective of the present study was to examine the effects of nano-selenium on growth performance, antioxidative status, and immune function in broiler chickens reared under thermoneutral (22±1°C) or high ambient temperature (35±1°C) conditions. Thirty-six broiler chicks at 15d old were randomly divided into 6 treatments in a 3×2 factorial design. The main factors included the dietary supplementation (basal diet without Se supplementation [control], basal diet with 0.3 mg of nano-elemental Se per kilogram of diet [nano-Se], and basal diet with 0.3 mg of sodium selenite per kilogram of diet [SSe]) and the ambient temperature challenge (22±1°C or 35±1°C). The birds were given the experimental diets from 15 to 30 d of age. High ambient temperature significantly depressed body weight gain, feed intake, feed conversion ratio, breast muscle weight, and abdominal fat weight, while feeding nano-Se clearly alleviated these negative effects of high ambient temperature. In addition, feeding nano-Se increased glutathione peroxidase mRNA expression in liver and alleviated the negative effects of high ambient temperature via reducing the malondialdehyde content in liver and breast muscle. Furthermore, feeding nano-Se increased mRNA expression of cytokine genes (interleukins 2 and 6) under both thermoneutral and high ambient temperature conditions. Under both thermoneutral and high-temperature conditions, broiler chickens fed nano-Se had higher Se and vitamin E concentrations in breast muscle than broiler chickens fed the control diet. In contrast, feeding SSe at the same dose as nano-Se did not alleviate the negative effects of high ambient temperature on broiler chickens. In conclusion, dietary supplementation with nano-Se at 0.3 mg/kg diet might enhance growth performance by improving antioxidative or immune properties in broilers reared under high ambient temperature.

A method of quantitative assessment of cytokine gene mRNA expression levels using real-time PCR in homogenous low cell number populations

We propose a method of quantitative functional activity assessment in cells isolated via sorting on a flow cytometer. We show that cell populations vary in the mRNA expression of cytokine genes immediately after isolation and sorting, while the maintenance of homogenous populations in culture without stimulation results in an increase in these gene mRNA expression. Using the original system, it is now possible to detect mRNA cytokine genes with high sensitivity, starting from 90 cells per specimen. This approach permits genetic and immunogenetic analysis of gene expression with the goal of determining their functions in the in vitro studies.

Cytokines in the colon of a patient with Behçet's disease

We read with interest the article by Canete and colleagues [1] in a recent issue of Arthritis Research & Therapy, in which they describe cytokine patterns of inflamed joints from patients with Behcet's disease (BD). The authors show that neutrophils and T lymphocytes are involved in a Th1-skewed inflammatory pattern expressed by elevations of interferon-gamma (IFN-γ), tumor necrosis factor-alpha (TNF-α), and interleukin-2 (IL-2). Also, increased levels of IL-4, IL-10, and IL-17 were observed. We would like to add to these observations our data showing a similar cytokine profile in the colon of a patient with BD. A 37-year-old BD patient with severe colitis failed to respond to traditional immunosuppressive treatment. Steroids were contraindicated because of a previous retinal serosal ablation, probably induced by prednisone. TNF blockage initially successfully reduced the severity of the colitis, but relapses occurred. Variation of different TNF blockers (etanercept, infliximab, and adalimumab) could not permanently resolve the intestinal complaints. Antibodies against infliximab or adalimumab were not detected. Eventually, high-dose infliximab (10 mg/kg) and intravenous immunoglobulins (IVIGs) led to disease regression facilitating a hemicolectomy. Thereafter, the patient's condition improved significantly and IVIGs were terminated while TNF blockage was continued. Cytokines were evaluated in the resected colon by analyzing mRNA expression levels of cytokine genes as previously described [2]. Because of the extensive prior treatment, the patient served as an internal control. Intestinal inflammation was patchy, and the observed difference between healthy and diseased tissues was confirmed microscopically. The mRNA expression of cytokines in diseased colon is presented relative to that of healthy colon and can be seen as a reflection of intramural cytokines (Figure ​(Figure1).1). Like Canete and colleagues, we observed a Th1- and Th17-skewed pattern (elevated IFN-γ and TNF-α and elevated IL-17A, respectively). We could not demonstrate elevation of IL-10, possibly because our patient was intensively treated and the colon environment is not sterile. Figure 1 Relative mRNA expression of cytokine genes in affected colonic tissue of a patient with Behcet's disease. Diseased colonic tissue and healthy colonic tissue from the same patient were analyzed. IFN-γ, interferon-gamma; IL, interleukin; ... The immunopathology of BD remains fascinating and highly relevant to the development of future immune-directed therapy. Since the successful introduction of TNF blockers, it became apparent that Th1-cytokines might be key players in the immunopathology of BD, emphasizing the importance of cytokine studies [3,4]. Canete and colleagues [1] have highlighted the importance of tissue evaluation in cytokine studies. In this light, our data add to their observations that, in colonic tissue of a BD patient, IFN-γ, TNF-α, and IL-17A appear to be key cytokines, even in a treated patient.

Potentially probiotic bacteria induce efficient maturation but differential cytokine production in human monocyte-derived dendritic cells

To analyze the ability of nine different potentially probiotic bacteria to induce maturation and cytokine production in human monocyte-derived dendritic cells (moDCs). Cytokine production and maturation of moDCs in response to bacterial stimulation was analyzed with enzyme-linked immunosorbent assay (ELISA) and flow cytometric analysis (FACS), respectively. The kinetics of mRNA expression of cytokine genes was determined by Northern blotting. The involvement of different signaling pathways in cytokine gene expression was studied using specific pharmacological signaling inhibitors. All studied bacteria induced the maturation of moDCs in a dose-dependent manner. More detailed analysis with S. thermophilus THS, B. breve Bb99, and L. lactis subsp. cremoris ARH74 indicated that these bacteria induced the expression of moDC maturation markers HLA class II and CD86 as efficiently as pathogenic bacteria. However, these bacteria differed in their ability to induce moDC cytokine gene expression. S. thermophilus induced the expression of pro-inflammatory (TNF-alpha, IL-12, IL-6, and CCL20) and Th1 type (IL-12 and IFN-gamma) cytokines, while B. breve and L. lactis were also potent inducers of anti-inflammatory IL-10. Mitogen-activated protein kinase (MAPK) p38, phosphatidylinositol 3 (PI3) kinase, and nuclear factor-kappa B (NF-kappaB) signaling pathways were shown to be involved in bacteria-induced cytokine production. Our results indicate that potentially probiotic bacteria are able to induce moDC maturation, but their ability to induce cytokine gene expression varies significantly from one bacterial strain to another.

Absence of Proinflammatory Cytokine Gene Expression in Nasal Biopsies from Wood Surface-coating Industry Workers

Symptoms of nasal, pharyngeal and ocular discomfort have been reported among workers in the wood surface-coating industry. Symptoms were reported more often by workers using ultraviolet radiation-curable acrylate coatings (UV coatings), which contain potential chemical sensitizers, than by those using acid-curing coatings. Furthermore, increased levels of eosinophil cationic protein (ECP) and albumin, but not tryptase, in nasal lavage from workers exposed to UV coatings have been observed. To further examine whether air contaminants present in the UV-coating industry are causing the observed increase in symptoms, the inflammatory process in the nasal mucosa of workers exposed to UV coatings was investigated. Clinical and biochemical endpoints were selected to distinguish between specific and non-specific hypersensitivity and to test the hypothesis that the symptoms were consistent with Type IV hypersensitivity. The nasal lavage and nasal biopsy were performed under local anesthetic at the workplace during working hours after a minimum of 2 h of work in both the exposed and control groups. Albumin and ECP, and the cytokines tumor necrosis factor- f (TNF- f ) and interleukin-8 (IL-8), were used as inflammatory markers. A multi-probe ribonuclease protection assay was used to attempt to detect cytokine variation in human nasal biopsies. The cytokine genes analyzed were TNF- f , GM-CSF, interferon- n , IL-2, IL-4 and IL-5. L32 and GAPDH were used as control genes for mRNA expression levels. Mucosal inflammation symptoms correlated with increased levels of albumin, but not with increased levels of ECP, secreted proinflammatory cytokines or cytokine gene mRNA expression. We conclude that the symptoms are non-specific and do not correlate with occupational exposure to UV coatings under the conditions of this investigation.

Interleukin 10 inhibits cytokine production of human AML cells

Unlike normal hematopoietic cells leukemic blasts from patients with AML constitutively express cytokines like IL1, GM-CSF and TNF alpha and it has been suggested that these cytokines may regulate growth and differentiation of the malignant cells. IL10 inhibits cytokine production of activated macrophages and T-helper 1 cells. We analysed whether IL10 can also suppress cytokine production and may inhibit growth of AML cells. AML blasts of 18 patients were purified by immunomagnetic separation and cultured in serum-free medium in the presence of cytokines. The production of cytokines was analysed by ELISA, DNA synthesis by 3H-thymidine incorporation and mRNA expression of cytokine genes by semiquantitative RT-PCR. Our results confirm previous results that AML blasts produce a variety of cytokines such as GM-CSF, IL1 alpha, IL1 beta, IL6 and TNF alpha. AML cells were induced to proliferation by G-CSF, GM-CSF, IL3, IL1 beta and SCF to a different extent. In contrast, IL10 significantly inhibited the cytokine production at the mRNA and protein level and spontaneous thymidine uptake in a dose-dependent way. This inhibition could be abrogated by IL10 specific antibodies. These observations suggest an inhibitory effect of IL10 on the proliferation of cultured AML blasts most likely through suppression of endogenous cytokines.