IL-1 IL-6 Research Articles

Constitutive and LPS-stimulated secretome of porcine Vascular Wall-Mesenchymal Stem Cells exerts effects on in vitro endothelial angiogenesis

BackgroundMSCs secretome is under investigation as an alternative to whole-cell-based therapies, since it is enriched of bioactive molecules: growth factors, cytokines and chemokines. Taking into account the translational value of the pig model, the leading aim of the present paper was to characterize the secretome of porcine Vascular Wall–Mesenchymal Stem Cells (pVW-MSCs) and its change in presence of LPS stimulation. Moreover, considering the importance of angiogenesis in regenerative mechanisms, we analysed the effect of pVW-MSCs secretome on in vitro angiogenesis.ResultsOur results demonstrated that conditioned medium from unstimulated pVW-MSCs contained high levels of IL-8, GM-CSF, IFN-γ and other immunomodulatory proteins: IL-6 IL-18 IL-4 IL-2 IL-10. LPS modulates pVW-MSCs gene expression and secretome composition, in particular a significant increase of IL-6 and IL-8 was observed; conversely, the amount of GM-CSF, IFN-γ, IL-2, IL-4, IL-10 and IL-18 showed a significant transient decrease with the LPS stimulation. Conditioned medium from unstimulated pVW-MSCs induced in vitro endothelial angiogenesis, which is more evident when the conditioned medium was from LPS stimulated pVW-MSCs.ConclusionsThe lines of evidence here presented shed a light on possible future application of secretome derived by pVW-MSCs on research studies in translational regenerative medicine.

The Role of the Tumor Microenvironment in Neuropilin 1-Induced Radiation Resistance in Lung Cancer Cells.

Background: Neuropilin 1 (NRP1) is a pleiotropic receptor which can interact with multiple ligands and their receptors. It plays an important role in the process of axonal growth, angiogenesis, tumor metastasis and radiation resistance in endothelial cells and some tumor cells. Interaction of stromal and tumor cells plays a dynamic role in initiating and enhancing carcinogenesis, and has received considerable attention in recent years.Material and Methods: In this study, A549 lung cancer cell lines with different NRP1 expression levels were constructed in vitro, a two-dimensional (2D), three-dimensional (3D) co-culture system and tumor-bearing model was established in SCID mice. Western blot, qRT-PCR, immunofluorescence, cytometric bead array and flow cytometry were used to investigate the effect of the tumor microenvironment in NRP1-induced lung cancer cell radiation resistance.Results: In 2D or 3D co-culture system, NRP1 could be regulated inflammatory factors such as TNF, IL-6 IL-8 and IL-17 and the related chemokines MCP-1, IP-10 and RANTES in the tumor microenvironment, which in turn induced radiation resistance in lung cancer cells. In addition, different expression levels of NRP1 in 2D, 3D culture systems and tumor-bearing models were able to significantly regulate cell phenotype, proliferative capacity, epithelial-mesenchymal transition (EMT) and the radiation resistance of A549 cells.Conclusion: Our results verified that NRP1, inflammatory factors, chemokines and related signaling pathways, which affect the transformation of related cell components and thus lung cancer cell immune tolerance and migratory ability, all play an important role in radiation resistance.

Severity of local inflammation does not impact development of fibrosis in mouse models of intestinal fibrosis

Intestinal fibrosis is thought to be a consequence of excessive tissue repair, and constitutes a common problem in patients with Crohn’s disease (CD). While fibrosis seems to require inflammation as a prerequisite it is unclear whether the severity or persistence of inflammation influences the degree of fibrosis. Our aim was to investigate the role of sustained inflammation in fibrogenesis. For the initiation of fibrosis in vivo the models of Il10−/− spontaneous colitis, dextran sodium sulfate (DSS)-induced chronic colitis and heterotopic transplantation were used. In Il10−/− mice, we determined a positive correlation between expression of pro-inflammatory factors (Il1β, Tnf, Ifnγ, Mcp1 and Il6). We also found a positive correlation between the expression of pro-fibrotic factors (Col3a1 Col1a1, Tgfβ and αSma). In contrast, no significant correlation was determined between the expression of pro-inflammatory Tnf and pro-fibrotic αSma, Col1a1, Col3a1, collagen layer thickness and the hydroxyproline (HYP) content. Results from the DSS-induced chronic colitis model confirmed this finding. In the transplantation model for intestinal fibrosis a pronounced increase in Mcp1, inos and Il6 in Il10−/− as compared to WT grafts was observed, indicating more severe inflammation in Il10−/− grafts. However, the increase of collagen over time was virtually identical in both Il10−/− and WT grafts. Severity of inflammation during onset of fibrogenesis did not correlate with collagen deposition. Although inflammation might be a pre-requisite for the initiation of fibrosis our data suggest that it has a minor impact on the progression of fibrosis. Our results suggest that development of fibrosis and inflammation may be disconnected. This may be important for explaining the inefficacy of anti-inflammatory treatments agents in most cases of fibrotic inflammatory bowel diseases (IBD).

Association between single nucleotide polymorphism of immunoregulatory genes and preterm premature rupture of membranes in preterm labour

Aim. Investigation of the association between SNP genes of IL4 (rs2243250), IL10 (rs1800896 and rs1800872) and RLN2 (rs4742076 and rs3758239) and preterm premature rupture of membranes in 26–34 weeks of gestation in Zaporizhzhia population. Materials and methods. W e have investigated markers of cytokines genes in 50 women with PPROM in 26–34 weeks of gestation period and 50 pregnant women with physiological pregnancy and term labour without complications. The genotyping using TaqMan tests was done on amplifier CFX96™ Real-Time PCR Detection Systems (Bio-Rad Laboratories, Inc., USA. The combined effect of the studied locus of the analyzed genes on the appearance of PPROM, PTL in the population was initiated using the Multifactory Dimensionality Reduction (MDR) method (software MDR ver. 3.0.2). Results. The distribution of the rs2243250 gene polymorphism alleles of the IL4 gene of the main study group – TT homozygotes were determined in 4 % cases, CT heterozygotes were found in 22 %, CC homozygotes – in 74 % cases. We have got statistically significant differences of rs4742076 polymorphisms (TT, CT, and CC) of the RLN2 gene in the study groups. We also detected statistically significant differences in all the alleles of rs3758239 polymorphism (AA, GG and AG) of the gene RLN2 – χ 2 = 23.86, OR( AA ) = 12.57; 95 % CI: 3.68–42.98; OR( GG )= 0.08; 95 % CI: 0.02–0.27; and OR( AG )= 0.11; 95 % CI: 0.03–0.42, respectively, P < 0.05, indicating the reliability of the received prognostic markers. The nature of interlocal interaction between the genes is at the level of “independent effects” of influence (IL4-IL10 rs1800896 = -23.08 %, IL10 rs1800896 – IL10 rs1800872 = -19.94 %, IL10 rs1800872-IL-4 = -19.34 %). The percentage of entropy of the investigated polymorphism of each gene for the case-control status was 14.15 % for the IL4 gene, 23.08 % for the IL10 rs1800896 gene and 9.34 % for the IL10 rs1800872 gene. Conclusions. Combination of IL4 rs2243250, IL10 rs1800896 and rs1800872 supports the role for functional polymorphisms in immunoregulatory genes in the development of PPROM and PTL. Reliable clinical association of rs4742076 and rs3758239 gene RLN2 was established with the PPROM in 26–34 weeks’ gestation in Zaporizhzhia population.

INCREASED LEVELS OF INTERLEUKIN-10 EXPRESSION COMPARED TO INTERLEUKIN-6 IN LEUKOCYTES OF HEALTHY SUBJECTS. COULD IT BE USEFUL IN THE FUTURE OF THE DEPRESSION DIAGNOSIS?

It is a well known, that the physical activity promotes mental health. Physically active people relatively rarely suffer from psychosomatic and depressive disorders. It is possible that the differences in gene activity in peripheral blood leukocytes may be associated with depression, especially genes participating in inflammatory response. Thus, the aim of our study was to investigate the levels of IL6 and IL10 mRNA and IL10/IL6 ratio in peripheral blood leukocytes in healthy, physically active individuals. One hundred healthy young men (20-23 years old) participated in this study. All of them declared regular physical activity. Participants were non-smokers, and consumed alcohol occasionally. To access genes expression, 2 ml of venous blood was collected. RNA isolation was performed and then the relative expression of IL6 and IL10 was calculated using quantitative real-time polymerase chain reaction (Q-RT-PCR). Pro and anti-inflammatory balance was calculated as 2^relative of IL10/ 2^relative expression of IL6. Low expression of tested genes was found in healthy young men. Mean expression for IL6 was 2^0.051 (n=90) and for IL10 2^0.08 (n=98). Mean ratio IL10/IL6 was 2^1.58. Higher expression of IL10 compared to IL6 may be essential not only for the physical performance but also for the mental health. Diverse reports in the literature may be associated with choosing various control groups, i.e. sedentary or older individuals. It is possible, that measuring the expression of IL6 and IL10 (especially the ratio IL10/IL6) in peripheral blood leukocytes may be useful in the assessment of depressive disorders. Thus, molecular study of active young men can confirm the need for physical activity among people suffering from depression, but further studies are needed, particularly among people with psychosomatic disorders.

Role of cytokines and inflammation in heart function during health and disease.

By virtue of their actions on NF-κB, an inflammatory nuclear transcription factor, various cytokines have been documented to play important regulatory roles in determining cardiac function under both physiological and pathophysiological conditions. Several cytokines including TNF-α, TGF-β, and different interleukins such as IL-1 IL-4, IL-6, IL-8, and IL-18 are involved in the development of various inflammatory cardiac pathologies, namely ischemic heart disease, myocardial infarction, heart failure, and cardiomyopathies. In ischemia-related pathologies, most of the cytokines are released into the circulation and serve as biological markers of inflammation. Furthermore, there is an evidence of their direct role in the pathogenesis of ischemic injury, suggesting cytokines as potential targets for the development of some anti-ischemic therapies. On the other hand, certain cytokines such as IL-2, IL-4, IL-6, IL-8, and IL-10 are involved in the post-ischemic tissue repair and thus are considered to exert beneficial effects on cardiac function. Conflicting reports regarding the role of some cytokines in inducing cardiac dysfunction in heart failure and different types of cardiomyopathies seem to be due to differences in the nature, duration, and degree of heart disease as well as the concentrations of some cytokines in the circulation. In spite of extensive research work in this field of investigation, no satisfactory anti-cytokine therapy for improving cardiac function in any type of heart disease is available in the literature.

FEATURES OF CYTOKINE REGULATION IN PATIENTS WITH MECHANICAL JAUNDICE OF MALIGNANT GENESIS

Mechanical jaundice (MJ) of malignant genesis is a clinical syndrome caused by obturation of bile ducts due to malignant tumor growth. It is characterized by specific pigmentation of skin, mucous membranes and sclerae due to increased accumulation of bilirubin in blood serum and other fluids and tissues of the organism. Various immune disorders are detected in patients with mechanical jaundice. Immune reactivity in mechanical jaundice is affected by toxic, infectious, or tumor factors. In connection with these findings, the aim of our study was to measure levels of some pro-inflammatory (IL-2, IL-18, TNFα, IFNγ) and antiinflammatory (IL-4 IL-10) cytokines in patients with malignant disease, depending on serum bilirubin levels. The study included 50 patients with MJ by malignant origin. A control group consisted of 125 practically healthy volunteers comparable with the study group by sex and age. Blood serum cytokine contents in patients and controls was determined by enzyme immunoassay using reagent kits produced by ZAO Vector-Best. Statistical evaluation of data was carried out by means of Statistica for Windows 8.0 and Microsoft Excel software. The levels of IL-2, TNFα, IFNγ and IL-18 were increased before surgery in the patients with mechanical jaundice of malignant genesis, while IL-4 and IL-10 values were reduced in comparison with healthy controls. Altered cytokine regulation in the MJ group manifests as increase in proinflammatory and immunostimulatory cytokine levels, along with decrease in anti-inflammatory cytokine amounts. Predominant feature of immune response may be regarded as prevalence of Th1-immune response when taking into account cytokine levels in blood serum of patients with MJ of malignant genesis. The features of cytokine regulation of patients with malignant MJ depend on the bilirubin content in blood serum. These changes were most frequent in patients with bilirubin level exceeding 60 μmol/L. In general, the quantitative indices of cytokines exhibited unidirectional changes in the groups of MJ patients with different bilirubin levels, except of IL-10. This parameter showed almost 6-fold increase in MJ patients with bilirubin level &lt; 60 μmol/L against control group, and approximately 40 times with respect to the remaining 2 groups. This biological effect may occur due to malignant etiology of the MJ.

Immune profile in aborted Iraqi women with toxoplasmosis

Background: Toxoplasmosis is one of the most important causes for abortion in women. The immune responses have a role in the outcome of such infection in gestated women. Aim: The current study was designed to investigate the immune profile in aborted Iraqi women with toxoplasmosis. Materials and Methods: Fifty-five aborted women and 29 healthy control women were enrolled in this study. Enzyme-linked immunosorbent assays were used to estimate serum levels to each of interleukin-1 alpha (IL-1α), granulocyte-monocyte colony-stimulating factor, IL-8, IL-4 IL-10, IL-12, interferon gamma (INF-γ), and IL-6. Single-radial-immunodiffusion assay was used to estimate serum levels of C3, C4, and total immunoglobulin gamma. Results: Serum levels of IL-8 showed significant elevation, while IL-6 and INF-γ showed significant dropping in infected women compared to control. Other immune factors showed nonsignificant differences between the two groups of the present study. Conclusion: Disturbance of immune response associated with toxoplasmosis may explain the success of parasite in escaping from discrimination and elimination by the immune system then supporting its survival and replication.

Using blood cytokine measures to define high inflammatory biotype of schizophrenia and schizoaffective disorder

BackgroundIncreases in pro-inflammatory cytokines are found in the brain and blood of people with schizophrenia. However, increased cytokines are not evident in all people with schizophrenia, but are found in a subset. The cytokine changes that best define this subset, termed the “elevated inflammatory biotype”, are still being identified.MethodsUsing quantitative RT-PCR, we measured five cytokine mRNAs (IL-1β, IL-2 IL-6, IL-8 and IL-18) from peripheral blood of healthy controls and of people with schizophrenia or schizoaffective disorder (n = 165). We used a cluster analysis of the transcript levels to define those with low and those with elevated levels of cytokine expression. From the same cohort, eight cytokine proteins (IL-1β, IL-2, IL-6, IL-8, IL-10, IL-12, IFNγ and TNFα) were measured in serum and plasma using a Luminex Magpix-based assay. We compared peripheral mRNA and protein levels across diagnostic groups and between those with low and elevated levels of cytokine expression according to our transcription-based cluster analysis.ResultsWe found an overall decrease in the anti-inflammatory IL-2 mRNA (p = 0.006) and an increase in three serum cytokines, IL-6 (p = 0.010), IL-8 (p = 0.024) and TNFα (p < 0.001) in people with schizophrenia compared to healthy controls. A greater percentage of people with schizophrenia (48%) were categorised into the elevated inflammatory biotype compared to healthy controls (33%). The magnitude of increase in IL-1β, IL-6, IL-8 and IL-10 mRNAs in people in the elevated inflammation biotype ranged from 100 to 220% of those in the non-elevated inflammatory biotype and was comparable between control and schizophrenia groups. Blood cytokine protein levels did not correlate with cytokine mRNA levels, and plasma levels of only two cytokines distinguished the elevated and low inflammatory biotypes, with IL-1β significantly increased in the elevated cytokine control group and IL-8 significantly increased in the elevated cytokine schizophrenia group.ConclusionsOur results confirm that individuals with schizophrenia are more likely to have elevated levels of inflammation compared to controls. We suggest that efforts to define inflammatory status based on peripheral measures need to consider both mRNA and protein measures as each have distinct advantages and disadvantages and can yield different results.

Betulinic acid derivative BA5, a dual NF-kB/calcineurin inhibitor, alleviates experimental shock and delayed hypersensitivity

Betulinic acid (BA) is a naturally occurring triterpenoid with several biological properties already described, including immunomodulatory activity. Here we investigated the immunomodulatory activity of eight semi-synthetic amide derivatives of betulinic acid. Screening of derivatives BA1-BA8 led to the identification of compounds with superior immunomodulatory activity than BA on activated macrophages and lymphocytes. BA5, the most potent derivative, inhibited nitric oxide and TNFα production in a concentration-dependent manner, and decreased NF-κB activation in Raw 264.7 cells. Additionally, BA5 inhibited the proliferation of activated lymphocytes and the secretion of IL-2, IL-4 IL-6, IL-10, IL-17A and IFNɣ, in a concentration-dependent manner. Flow cytometry analysis in lymphocyte cultures showed that treatment with BA5 induces cell cycle arrest in pre-G1 phase followed by cell death by apoptosis. Moreover, BA5 also inhibited the activity of calcineurin, an enzyme that plays a critical role in the progression of cell cycle and T lymphocyte activation. BA5 has a synergistic inhibitory effect with dexamethasone on lymphoproliferation, showing a promising profile for drug combination. Finally, we observed immunosuppressive effects of BA5 in vivo in mouse models of lethal endotoxemia and delayed type hypersensitivity. Our results reinforce the potential use of betulinic acid and its derivatives in the search for potent immunomodulatory drugs.

MCL-1 is modulated in Crohn\u2019s disease fibrosis by miR-29b via IL-6 and IL-8

The miR-29 family is involved in fibrosis in multiple organs, including the intestine where miR-29b facilitates TGF-β-mediated up-regulation of collagen in mucosal fibroblasts from Crohn’s disease (CD) patients. Myeloid cell leukemia-1 (MCL-1), a member of the B-cell CLL/Lymphoma 2 (BCL-2) apoptosis family, is involved in liver fibrosis and is targeted by miR-29b via its 3’-UTR in cultured cell lines. We investigate the role of MCL-1 and miR-29b in primary intestinal fibroblasts and tissue from stricturing CD patients. Transfection of CD intestinal fibroblasts with pre-miR-29b resulted in a significant increase in the mRNA expression of MCL-1 isoforms [MCL-1Long (L)/Extra Short (ES) and MCL-1Short (S)], although MCL-1S was expressed at significantly lower levels. Western blotting predominantly detected the anti-apoptotic MCL-1L isoform, and immunofluorescence showed that staining was localised in discrete nuclear foci. Transfection with pre-miR-29b or anti-miR-29b resulted in a significant increase or decrease, respectively, in MCL-1L foci. CD fibroblasts treated with IL-6 and IL-8, inflammatory cytokines upstream of MCL-1, increased the total mass of MCL-1L-positive foci. Furthermore, transfection of intestinal fibroblasts with pre-miR-29b resulted in an increase in mRNA and protein levels of IL-6 and IL-8. Finally, immunohistochemistry showed reduced MCL-1 protein expression in fibrotic CD samples compared to non-stricturing controls. Together, our findings suggest that induction of MCL-1 by IL-6/IL-8 may surmount any direct down-regulation by miR-29b via its 3’-UTR. We propose that an anti-fibrotic miR-29b/IL-6 IL-8/MCL-1L axis may influence intestinal fibrosis in CD. In the future, therapeutic modulation of this pathway might contribute to the management of fibrosis in CD.

Effect of IL-1β or TNF-α-treated human decidual cells on balance between Th1 and Th2 or Th17 and Treg immunity

To investigate the effect of IL-1β or TNF-α-treated human decidual cells on the balance between Th1 and Th2 or Th17 and Treg. Methods: Human decidual cells were isolated and treated with estrogen (E2) and progesterone (P) for 7 days and then with IL-1β or TNF-α for 24 h. The culture medium was collected to prepare conditioned medium. The T cells isolated from human decidual tissue were cultured in presence of conditioned medium for 72 h. ELISA was used to detect concentration of IFN-γ, IL-2, IL-5, IL-17 IL-10, and TGF-1β in supernatants and quantitative real-time PCR was used to detect the mRNA expression of IFN-γ, IL-17, IL-10, and TGF-1β. Results: The levels of IFN-γ, IL-2 and IL-17 were significantly increased when T cells were cultured in the conditioned medium compared with the control group (E2+P), while the levels of IL-5, IL-10, and TGF-1β were not significantly changed. The mRNA expressions of IFN-γ and IL-17 mRNA were significantly increased when T cells were cultured in the conditioned medium compared with the control group (E2+P), while the mRNA expression of IL-10 and TGF-1β was not significantly different. Conclusion: Human decidual cells treated by IL-1β or TNF-α can shift the balance of Th1 and Th2 or Th17 and Treg toward Th1 and Th17 immunity.

Phagocytosis and immune response studies of Macrophage-Nanodiamond Interactions in vitro and in vivo.

The applications of nanodiamond as drug delivery and bio-imaging can require the relinquishing ND-drug conjugate via blood flow, where interaction with immune cells may occur. In this work, we investigated the ND penetration in macrophage and the immune response using the tissue-resident murine macrophages (RAW 264.7). Confocal fluorescence imaging, immunofluorescence analysis of nuclear translocation of interferon regulatory factor IRF-3 and transcriptional factor NF-κΒ, analysis of pro-inflammatory cytokines production IL-1β, IL-6 IL-10 with a reverse transcription-polymerase chain reaction technique were applied. The TNF-α factor production has been studied both in vitro at ND interaction with the macrophage and in vivo after ND injection in the mice blood system using immunoassay. The macrophage antibacterial function was estimated through E. coli bacterial colony formation. ND didn't stimulate the immune response and functionality of the macrophage was not altered. Using MTT test, ND was found negligibly cytotoxic to macrophages. Thus, ND can serve as a biocompatible platform for bio-medical applications. Left: Graphic representation of Nanodiamond internalization in macrophage. Right: (a) Fluorescence images of lysosomes, (b) nanodiamond and (c) merged image of nanodiamond internalization in macrophage.

A Human 3D In Vitro Model to Assess the Relationship Between Osteoporosis and Dissemination to Bone of Breast Cancer Tumor Cells.

Despite consistent improvements in diagnostic and therapeutic strategies for breast cancer, up to 40% of patients will develop bone metastases. To reduce the morbidity and complications related with bone metastases, it is imperative to reduce their etiological factors. Osteoporosis, being characterized by a sudden estrogen deficiency, may provide a favorable condition for bone metastasis. This work, using a humanized 3D in vitro model, aims at evaluating the relationship between osteoporosis and breast cancer-derived bone metastases. Bone tissue discarded from total hip replacement surgery of healthy and osteoporotic patients was cultured in a rolling apparatus system in hypoxic environment. Protein levels (i.e., vascular endothelial growth factor (VEGF), VEGF receptor 1, VEGF receptor 2, interleukin (IL)-6, IL-1β, IL-8 IL-10, tumor necrosis factor α (TNF-α), osteoprotegerin (OPG), receptor activator for nuclear factor KB ligand (RANKL)) and histological and immunohistochemical (i.e., cytokeratin 8 and 18) analyses showed a noticeable specificity of breast cancer cells for the colonization of osteoporotic bone. These data are the first to demonstrate that using humanized 3D in vitro systems, which individually model the pre- and postmenopausal bone microenvironment, it is possible to recognize major differences in tumor growth and colonization between healthy and osteoporotic status. Thus, this system might help to develop a shared system between basic and clinical sciences where a personalized diagnosis is associated to a therapeutic strategy designed for a single patient: a model able to achieve a translational research approach in the clinical setting, which may lead to the application and dissemination of personalized medicine. J. Cell. Physiol. 232: 1826-1834, 2017. © 2016 Wiley Periodicals, Inc.

Pathogenic mechanisms of pancreatitis.

Pancreatitis is inflammation of pancreas and caused by a number of factors including pancreatic duct obstruction, alcoholism, and mutation in the cationic trypsinogen gene. Pancreatitis is represented as acute pancreatitis with acute inflammatory responses and; chronic pancreatitis characterized by marked stroma formation with a high number of infiltrating granulocytes (such as neutrophils, eosinophils), monocytes, macrophages and pancreatic stellate cells (PSCs). These inflammatory cells are known to play a central role in initiating and promoting inflammation including pancreatic fibrosis, i.e., a major risk factor for pancreatic cancer. A number of inflammatory cytokines are known to involve in promoting pancreatic pathogenesis that lead pancreatic fibrosis. Pancreatic fibrosis is a dynamic phenomenon that requires an intricate network of several autocrine and paracrine signaling pathways. In this review, we have provided the details of various cytokines and molecular mechanistic pathways (i.e., Transforming growth factor-β/SMAD, mitogen-activated protein kinases, Rho kinase, Janus kinase/signal transducers and activators, and phosphatidylinositol 3 kinase) that have a critical role in the activation of PSCs to promote chronic pancreatitis and trigger the phenomenon of pancreatic fibrogenesis. In this review of literature, we discuss the involvement of several pro-inflammatory and anti-inflammatory cytokines, such as in interleukin (IL)-1, IL-1β, IL-6, IL-8 IL-10, IL-18, IL-33 and tumor necrosis factor-α, in the pathogenesis of disease. Our review also highlights the significance of several experimental animal models that have an important role in dissecting the mechanistic pathways operating in the development of chronic pancreatitis, including pancreatic fibrosis. Additionally, we provided several intermediary molecules that are involved in major signaling pathways that might provide target molecules for future therapeutic treatment strategies for pancreatic pathogenesis.

T-cell redeployment and intracellular cytokine expression following exercise: effects of exercise intensity and cytomegalovirus infection.

The magnitude of lymphocytosis following exercise is directly related to exercise intensity. Infection with cytomegalovirus (CMV) also augments lymphocytosis after exercise. It is not known if the enhanced T‐cell response to exercise due to CMV depends on exercise intensity. Furthermore, exercise‐induced changes in T‐cell expression of type I and type II cytokines are thought to be intensity dependent, but direct comparisons are lacking. The aim of this experiment was to determine if CMV affects the exercise‐induced redistribution of T‐cell subsets at varying intensities, and determine the effect of exercise intensity on CD8+ T‐cell cytokine expression. Seventeen cyclists (nine CMV seropositive; CMV+) completed three 30 min cycling trials at −5, +5, and +15% of blood lactate threshold (LT). T‐cell subsets in blood and intracellular expression of type I (IL‐2, interferon(IFN)‐γ) and type II (IL‐4, IL‐10) cytokines by CD8+ T cells pre, post, and 1‐h post‐exercise were assessed by flow cytometry. Independently of CMV, T‐cell subset redistribution was greater after +15%LT compared to −5%LT (P < 0.05). Independently of intensity, CMV− mobilized more low‐ (CD27+ CD28+) and medium‐ (CD27+ CD28−) differentiated T cells than CMV+, whereas CMV+ mobilized more high (CD27− CD28−) differentiated T cells. The numbers of IL‐2+, IFN‐γ+, IL‐4+, and IL‐10+ CD8+ T cells increased after exercise above LT. Only type I cytokine expression was influenced by exercise intensity (P < 0.05). In conclusion, T‐cell redeployment by exercise is directly related to exercise intensity, as are changes in the number of CD8+ T‐cells expressing type I cytokines. Although CMV+ mobilized more high‐differentiated T cells than CMV−, this occurred at all intensities. Therefore, the augmenting effect of CMV on T‐cell mobilization is independent of exercise intensity.

Sodium Butyrate Reduces Colitogenic Immunoglobulin A-Coated Bacteria and Modifies the Composition of Microbiota in IL-10 Deficient Mice.

High levels of immunoglobulin A (IgA)-coated bacteria may have a role in driving inflammatory bowel disease (IBD). We therefore investigated the effect of sodium butyrate on microbiota in IBD prone interleukin (IL)-10−/− mice. At 8 weeks of age, mice were allocated into three groups (n = 4/group): normal (C57BL/6), IL-10−/−, and IL-10−/− treated with sodium butyrate (100 mM). Severity of colitis, inflammatory cytokine and short-chain fatty acid (SCFA) concentration in proximal colon contents, the percentage of IgA-coated bacteria and microbiota composition by 16S ribosomal RNA assessment of stool were measured after 4 weeks of treatment. Sodium butyrate ameliorated histological colitis and decreased levels of tumor necrosis factor (TNF)-α and IL-6 in IL-10−/− mice compared with those without treatment. At the phylum level, a reduction in Bacteroidetes and an increase in Firmicutes in IL-10−/− mice treated with sodium butyrate were observed. Additionally, Prevotellaceae species were reduced in IL-10−/− mice treated with sodium butyrate as compared with those without treatment. The level of biodiversity was slightly increased and the amount of IgA-coated bacteria decreased in IL-10−/− mice treated with sodium butyrate compared with those without treatment. Our results indicate that sodium butyrate protects against colitis, possibly through modifying the gut microbiota, enriching biodiversity and reducing the amount of colitogenic IgA-coated bacteria in IL-10−/− mice.

Relationship between Inflammatory Cytokines and Indices of Cardiac Dysfunction following Intense Endurance Exercise

ObjectivesPro-inflammatory cytokines have been noted to increase following exercise but their relationship to exercise-induced cardiac dysfunction has not previously been investigated. We sought to evaluate whether exercise-induced cardiac dysfunction was associated with increases in cytokines, particularly the pro-inflammatory cytokines IL-1β, IL-12p70 and TNFα, which have been most implicated in cardiac pathology.Methods40 well-trained endurance athletes underwent evaluation prior to and immediately following one of four endurance sporting events ranging from 3 to 11 hours duration. Cytokines (IL-1β, IL-6, IL-8, IL-10, IL-12p70 and TNFα) were analyzed by flow cytometry from serum samples collected within 50 minutes of race completion. Cardiac troponin (cTnI) and B-type natriuretic peptide were combined with an echocardiographic assessment of cardiac function, and a composite of cTnI > 0.04 μg/L, BNP increase > 10 ng/L and a decrease in right ventricular ejection (RVEF) > 10% were prospectively defined as evidence of myocardial dysfunction.ResultsRelative to baseline, IL-6 IL-8 and IL-10 increased 8.5-, 2.9-, and 7.1-fold, respectively, P<0.0001. Thirty-one (78%), 19 (48%) and 18 (45%) of the athletes met the pre-specified criteria for significant cTnI, BNP and RVEF changes, respectively. TNFα, IL-12p70 were univariate predictors of ΔRVEF and ΔBNP whilst none of the anti-inflammatory cytokines were significantly associated with these measures. Ten athletes (25%, all athletes competing in the endurance event of longest duration) met criteria for exercise-induced myocardial dysfunction. In these 10 athletes with myocardial dysfunction, as compared to those without, there was significantly greater post-race expression of the pro-inflammatory cytokines IL-12p70 (8.1±3.8pg/ml vs. 2.5±2.6pg/ml, P<0.0001) and TNFα (6.5±3.1pg/ml vs. 2.0±2.5pg/ml, P<0.0001).ConclusionCardiac dysfunction following intense endurance exercise was associated with increased expression of pro-inflammatory cytokines. This does not prove a causal relationship but provides rationale for further investigations into whether inflammation mediates exercise-induced myocardial dysfunction.

English

Objectives: To determine the effects of Epigallocatechin Gallate (EGCG), a major catechin component of green tea, on cytokine expression in a human oral epithelial cell culture model of nicotine use. Methods: Confluent gingival epithelial cells in wells of a 24-well plate were subjected to one of six treatments. For controls cells received 1) no treatment or 2) were treated with 10 μg /ml EGCG for 1 hour and cultured for 24 hours prior to analysis. A set of cells were pre-treated for 1 hour with 10 μg /ml EGCG and 3) treated for 24 hours with 0.1 mM nicotine prior to challenge with 10 ng/ml TNFα for 1 hour, or 4) not treated with nicotine but challenged with TNFα for 1 hour prior to analysis. A set of cells were not pre-treated with EGCG and 5) treated for 24 hours with 0.1 mM nicotine prior to challenge with 10 ng/ml TNFα for 1 hour, or 6) not treated with nicotine but challenged with TNFα for 1 hour prior to analysis. Culture medium samples were assayed for levels of secreted interleukins IL-1α, IL-1, IL-4, IL-6, IL-8, and IL-10 by ELISA. Statistical analysis was completed for individual interleukins using ANOVA and Tukey post-test with probability set at p ≤ 0.05. Results: The levels of IL-4 for all treatments were below the detection limit of the ELISA. EGCG significantly suppressed the secretion of IL-1α, IL-1IL-6 and IL-8 in nicotine and TNFα-treated cells (p < 0.01). In contrast, the presence of EGCG resulted in a significant increase in the secretion of IL-10 in nicotine and TNFα-treated cells (p < 0.001). Conclusion: This study suggests that tea catechins such as EGCG may function to suppress pro-inflammatory cytokines and to increase the secretion of the anti-inflam matory cytokine IL-10 in cells challenged by nicotine and TNFα.

Mo1734 Serum Fatty Acids Are Correlated With Tissue Cytokine Levels in Ulcerative Colitis

until used for cytokine determination. The concentration of intestine mucosa cytokines involved in the Th1 (IL-2, IL-12p70, IFN-γ), Th2 (IL-4), Th17 (IL-6, IL-17A, IL-23) and Treg (IL-10, TGF-β) cell commitment and IL-1 β and TNF-α (cytokines of the innate immune response) was assayed in duplicate by ELISA (Biosource, Camarillo, CA) and the results were expressed as picogram of cytokine per milligram of protein. Data were analyzed with SPSS. Comparisons between the groups were done by the two-tailed Mann-Whitney U-test. The ileal and colonic concentration of all proinflammatory cytokines was significantly higher in the patients with CD and also UC than in control group (p<0.001 for all). Otherwise, ileal IL-10 (369.7±47.9 vs. 1569.3±81.7) and TGF-β (10534.9±391.2 vs. 13045.8±1005.8) and colonic IL-10 (225.2±47.1 vs. 1563.3±81.7) and TGF-β (7224.0±306.7 vs. 12962.8±991.4) concentration was lower in UC than in controls. Also ileal IL-10 (224.9±23.9) and TGF-β (7073.6±54.4) and colonic IL-10 (310.7±55.3) and TGF-β (9089.1±1850.3) concentration was lower in CD patients than in controls. Significant differences between CD and UC were also observed. Thus, IL-2, IL-6, IL-17, IL-23 ileal and colonic concentration was higher in DC than in UC. Otherwise, IL-10 ileal concentration was lower in CD and IL-10 colonic concentration was lower in UC. The concentration of other cytokines including those of the innate immunity and Th1 response was higher in the ileum of CD and higher in the colon of UC when the diseases where compared. Next CD with and without complications were compared. Although no significant differences were observed between patients with stenosis (n=17) and fistula (n=7), the colonic concentration of Th17-associated cytokines and TNF-α was significantly higher (p<0.03), either in patients with stenosis or with fistula than in those without complications .In conclusion, in CD predominates a Th17 shift, and an insuficient Treg cell activation to suppress the overwhelming proinflammatory milieu was observed in both diseases.