Treatment Of Chronic Inflammatory Diseases Research Articles

Evidence of treatment of chronic inflammation in childhood and adolescence with biologics

Biologics, usually monoclonal antibodies or fusion proteins, are thought to specifically interfere with immunopathogenesis of chronic inflammatory diseases. In order to test these substances also in children and adolescents, financial incentives for manufacturers were created and classification of chronic inflammatory diseases and definition of disease activity, improvement, relapse and remission were established and large international research cooperation projects were founded. A selective literature search was carried out for treatment of chronic inflammatory diseases in children and adolescents with biologics including current guidelines. Only 7 out of 18 prescribed biologics have been approved for children and mostly within narrow limits. The evidence for efficacy is based on four randomized double blind placebo-controlled studies, seven withdrawal studies and seven observational studies. In spite of the limited evidence in comparison to their use in adult patients these substances are broadly used worldwide and have enlarged and substantially improved the therapeutic choices in children when conventional treatment failed or proved to be toxic. Severe adverse events including infections occasionally occur (0.01-0.03 events per patient year) but the rate of malignancies is not obviously increased; however, only two thirds of patients respond to treatment. Improvement is often incomplete, some patients deteriorate and definite termination of drug treatment is possible in only a few patients. As the prescription of biologics has become an important issue of treatment but is based on insufficient evidence data, further studies are necessary in children and adolescents with diseases, such as juvenile idiopathic arthritis, Crohn's disease, ulcerative colitis and inherited fever syndromes. As many drugs are available these studies can be conducted against verum.

Anti-inflammatory Effects of (-)-Epicatechin in Lipopolysaccharide-Stimulated Raw 264.7 Macrophages

Purpose: To investigate the protective effects of (-)-epicatechin on lipopolysaccharide (LPS)-induced inflammation in Raw 264.7 murine macrophages and the possible underlying mechanisms. Methods: The effects of epicatechin on LPS-stimulated production of inflammatory mediators in Raw 264.7 cells were evaluated by enzyme-linked immunosorbent assay. Results: Epicatechin in doses of 5, 25 and 50 µM remarkably (p < 0.05) inhibited the production of proinflammatory mediators including nitric oxide (NO) and prostaglandin E2 (PGE 2), as well as the production of pro-inflammatory cytokines including tumor necrosis factor (TNF)-alpha and interleukin (IL)-6 in LPS-induced Raw 264.7 macrophages. Conclusion: The results suggest that epicatechin can inhibit inflammatory response and may be a potential therapeutic candidate for the treatment of chronic inflammatory diseases.

Subcutaneous Merkel Cell Carcinoma in a Psoriatic Patient Treated with Methotrexate and TNF-α inhibitors

TNF-α inhibitors are used for the treatment of chronic inflammatory diseases and provide a high degree of clinical efficacy in patients with severe psoriasis. Although the effect of blocking agents gives significant improvements in quality of life of psoriatic patients, the use of them might lead to neoplasia not observed before the treatment. We present a case of Merkel cell carcinoma in a patient with a long-standing history of psoriasis and psoriatic arthritis treated with methotrexate and TNF-α inhibitors (etanercept and infliximab).

Mitogen-activated protein kinase-activated protein kinase 2 deficiency reduces insulin sensitivity in high-fat diet-fed mice.

Adipose tissue inflammation is considered an important contributor to insulin resistance. Mitogen-activated protein kinase-activated protein kinase 2 (MK2) is a major downstream target of p38 MAPK and enhances inflammatory processes. In line with the role of MK2 as contributor to inflammation, MK2−/− mice are protected against inflammation in different disease models. Therefore, MK2 is considered an attractive therapeutic target for the treatment of chronic inflammatory diseases. This study tested the impact of MK2-deficiency on high-fat diet (HFD)-induced adipose tissue inflammation and insulin resistance. After feeding MK2−/− and WT control mice a HFD (60% energy from fat) for 24 weeks, body weight was not different between groups. Also, liver weight and the amount of abdominal fat remained unchanged. However, in MK2−/− mice plasma cholesterol levels were significantly increased. Surprisingly, macrophage infiltration in adipose tissue was not altered. However, adipose tissue macrophages were more skewed to the inflammatory M1 phenotype in MK2−/− mice. This differerence in macrophage polarization did however not translate in significantly altered expression levels of Mcp-1, Tnfα and Il6. Glucose and insulin tolerance tests demonstrated that MK2−/− mice had a significantly reduced glucose tolerance and increased insulin resistance. Noteworthy, the expression of the insulin-responsive glucose transporter type 4 (GLUT4) in adipose tissue of MK2−/− mice was reduced by 55% (p<0.05) and 33% (p<0.05) on the mRNA and protein level, respectively, compared to WT mice. In conclusion, HFD-fed MK2−/− display decreased glucose tolerance and increased insulin resistance compared to WT controls. Decreased adipose tissue expression of GLUT4 might contribute to this phenotype. The data obtained in this study indicate that clinical use of MK2 inhibitors has to be evaluated with caution, taking potential metabolic adverse effects into account.

S-21 : Blocking only the bad side of Interleukin-6

Cytokines receptors exist in membrane bound and soluble form. The IL-6/soluble IL-6R complex stimulates target cells not stimulated by IL-6 alone, since they do not express the membrane bound IL-6R. We have named this process ‘ trans-signaling ’. The soluble IL-6R is generated via ectodomain shedding by the membrane bound metalloprotease ADAM17. Soluble gp130 is the natural inhibitor of IL-6/soluble IL-6R complex responses [1] . The dimerized recombinant soluble gp130Fc fusion protein is a molecular tool to discriminate between gp130 responses via membrane bound and soluble IL-6R responses. Interestingly, depending on the animal model used, global blockade of IL-6 signaling by neutralizing monoclonal antibodies and selective blockade of IL-6 trans-signaling can lead to different consequences. We used neutralizing monoclonal antibodies for global blockade of IL-6 signaling and the sgp130Fc protein for selective blockade of IL-6 trans-signaling in several animal models of human diseases. Inhibition of IL-6 trans-signaling but not global IL-6 blockade was beneficial in the cecal ligation and puncture sepsis model [2] . Defense against bacterial infections rely on the membrane bound IL-6R [3] . Acute pancreatitis often results in subsequent acute lung injury, which is an inflammatory disease with high mortality. IL-6 is necessary for the inflammatory process to reach the lung and blockade of IL-6 trans-signaling is sufficient to block the disease [4] . The extent of inflammation is controlled by trans-signaling via the soluble IL-6R. Using the sgp130Fc protein or sgp130Fc transgenic mice we demonstrate in animal models of inflammatory bowel disease, peritonitis, rheumatoid arthritis, atherosclerosis, pancreatitis, lupus erythematodes, colon cancer, ovarian cancer and pancreatic cancer that IL-6 trans-signaling via the soluble IL-6R is the crucial step in the development and the progression of the disease [1] , [2] , [3] , [4] , [5] . Therefore, sgp130Fc is a novel therapeutic agent for treatment of chronic inflammatory diseases and cancer and it underwent phase I clinical trials as an anti-inflammatory in 2013/2014. Phase II clinical trials in patients with autoimmune diseases such as inflammatory bowel disease will follow next year.

Vitamin D3 metabolite calcidiol primes human dendritic cells to promote the development of immunomodulatory IL-10-producing T cells

Vitamin D is recognized as a potent immunosuppressive drug. The suppressive effects of vitamin D are attributed to its physiologically active metabolite 1,25 dihydroxy vitamin D3 (calcitriol), which was shown, to prime dendritic cells (DCs) to promote the development of regulatory T (Treg) cells. Despite the potential benefit in treating autoimmune diseases, clinical application of calcitriol is hindered by deleterious side effects manifested by hypercalcemia and hypercalciuria. Conversely, the physiological precursors of calcitriol, vitamin D3 (cholecalciferol) and its first metabolite 25-hydroxy vitamin D3 (calcidiol) are widely applied in the clinic due to their low calcimic burden. However, the mechanisms by which cholecalciferol and calcidiol may modulate adaptive immunity remain elusive. This prompted us to unravel the immunosuppressive capacity of these precursors by assessing their influence on DC functions and the subsequent polarization of naïve CD4+ T cells. In this study we show that, whereas cholecalciferol has insignificant effects on DC maturation and cytokine production, it only weakly primed DCs to induce suppressive T cells. However, like calcitriol, calcidiol not only exerted an inhibitory effect on DC maturation and cytokine production, and primed DCs to promote the development of suppressive IL-10-producing Treg cells. Strikingly, in contrast to the population of IL-10-producing Treg cells induced by calcitriol-primed DCs, the IL-10-producing Treg cells induced by calcidiol-primed DCs exhibited sustained IFN-γ production in face of their suppressive capacity. Experiments with the steroid synthesis inhibitor ketoconazole indicated that the immunomodulatory features of the precursors are dependent on their conversion into calcitriol. Collectively, calcidiol is a potent immune modulator, which may be more adequate than calcitriol for the treatment of chronic inflammatory diseases, since it is less hypercalcimic. This may be of particular interest for the treatment of allergic disease, where concurrent suppression and sustained IFN-γ production by Treg cells effectively counterbalance the Th2-dominated immune responses.

Mechanisms of glucocorticoid action and insensitivity in airways disease

Glucocorticoids are the mainstay for the treatment of chronic inflammatory diseases including asthma and chronic obstructive pulmonary disease (COPD). However, it has been recognized that glucocorticoids do not work well in certain patient populations suggesting reduced sensitivity. The ultimate biologic responses to glucocorticoids are determined by not only the concentration of glucocorticoids but also the differences between individuals in glucocorticoid sensitivity, which is influenced by multiple factors. Studies are emerging to understand these mechanisms in detail, which would help in increasing glucocorticoid sensitivity in patients with chronic airways disease. This review aims to highlight both classical and emerging concepts of the anti-inflammatory mechanisms of glucocorticoids and also review some novel strategies to overcome steroid insensitivity in airways disease.

Phytochemical Constituents in Hexane Fraction of Costus afer Ker Gawl. Stem

In view of the high demands for alternative medicines in developing countries and the dearth of valid scientific data on herbal formulations. This study was designed to identify and characterize the bioactive compounds present in hexane fraction of Costus afer ker Gawl stem, an indigenous medicinal African plant used as therapy in the treatment of chronic inflammatory disease such as rheumatoid arthritis. Phytochemical evaluation and gas chromatography-mass spectrometry (GC/MS) analytical methods were carried out. The mass spectra of the identified compounds were compared with those of the National Institute of Standards and Technology database library. Results showed the presence of alkaloids, diterpenes, triterpenes, phytosterol, phenol, phlobatannins and tannins while GC/MS data detected 16 compounds including benzofuran, 2,3-dihydro and oleic acid. These compounds identified in the hexane fractions of C. afer stem may explain the folkloric use of C. afer plant stem extract in the treatment of chronic inflammatory diseases.

Immunoregulatory properties of Hsp70

Heat shock proteins 70 (Hsp70) play an important role in maintaining cellular homeostasis. As molecular chaperones, Hsp70 are responsible for proper folding of newly synthesized polypeptides and refolding of misfolded and aggregated proteins. Hsp70 are involved in cellular transport and participate in signal transduction processes. The presence of Hsp70 in the extracellular space is associated with development of various pathological conditions, including autoimmune and cancer diseases. Recent data suggest anti-inflammatory property of Hsp70 confirmed in both in vitro and animal models. This paper summarizes recent data regarding immunoregulatory properties of Hsp70, and presents promising results of the studies concerning the application of heat shock proteins in treatment of chronic inflammatory diseases.

Etanercept exacerbates inflammation and pathology in a rabbit model of active pulmonary tuberculosis.

Treatment of chronic inflammatory diseases with tumor necrosis factor alpha (TNF-α) antagonists has been associated with increased risk of tuberculosis (TB). We examined the usefulness of the rabbit model of active pulmonary TB for studying the impact of the human immune modulatory reagent etanercept on the host immune response. Control of Mycobacterium tuberculosis (Mtb) infection, disease pathology, and the global transcriptional response in Mtb-infected lungs of rabbits were studied. Etanercept treatment exacerbated disease pathology and reduced bacillary control in the lungs, compared with infected untreated animals. Reduced collagen and fibrin deposition in the granulomas was associated with significant downregulation of the collagen metabolism and fibrosis network genes and upregulation of genes in the inflammatory response and cell recruitment networks in the lungs of etanercept treated, compared with untreated rabbits. Our results suggest that targeting the TNF-α signaling pathway disrupts the tissue remodeling process, which is required for the formation and maintenance of well-differentiated granulomas and for control of Mtb growth in the lungs. These results validate the use of the rabbit model for investigating the impact of selected human immune modulatory drugs, such as a TNF-α antagonist, on the host immune response and pathogenesis in TB.

Sexually dimorphic actions of glucocorticoids: beyond chromosomes and sex hormones.

Sexual dimorphism is a well-documented phenomenon that is observed at all levels of the animal kingdom. Historically, sex hormones (testosterone and estrogen) have been implicated as key players in a wide array of pathologies displaying sexual dimorphism in their etiology and progression. While these hormones clearly contribute to sexually dimorphic diseases, other factors may be involved in this phenomenon as well. In particular, the stress hormone cortisol exerts differential effects in both males and females. The underlying molecular basis for the sexually dimorphic actions of glucocorticoids is unknown but clearly important to understand, since synthetic glucocorticoids are the most widely prescribed medication for the treatment of chronic inflammatory diseases and hematological cancers in humans.

The anti-inflammatory efficacy of dexamethasone is protected by (−)-epicatechin

Abstract Dexamethasone and other glucocorticoids (GCs) are used in the treatment of chronic inflammatory diseases. However, in diseases like chronic obstructive pulmonary disease GC resistance can develop. Our study examines (−)-epicatechin, a dietary compound known to display anti-inflammatory action, as adjuvant to prevent GC resistance. The anti-inflammatory potency of dexamethasone was determined in a carefully characterized system of monocytes differentiated to macrophage-like cells. The LPS induced IL-8 release was measured after pre-incubation with H 2 O 2 and (−)-epicatechin. We show that dexamethasone could efficiently suppress LPS induced inflammation and that exposure to H 2 O 2 induced GC resistance. The anti-inflammatory effect of dexamethasone was protected by (−)-epicatechin in the oxidant exposed cells. These results show that the natural antioxidant (−)-epicatechin may be used as an adjuvant therapy to restore or preserve the efficacy of GC drugs and thereby improving the treatment of inflammation.

Bioenergetic characterization of macrophage inflammatory responses attenuated by anthocyanins (383.2)

Anthocyanins present strong anti‐inflammatory properties, thus leading to the potential use as functional food ingredients for the prevention and treatment of chronic inflammatory diseases and increased intestinal permeability to endotoxins, notably LPS. However, the differences between individual anthocyanins and their effects on cellular signaling and metabolism need to be further elucidated. In this study, we evaluated the protection afforded by anthocyanins against LPS‐induced inflammatory response in RAW 264.7 macrophages and determined changes in oxygen consumption rates (OCR) and extracellular acidification rates (ECAR) during the macrophage activation. Basal OCR and ECAR increased as the macrophages underwent LPS stimulation from 283.3±38.4 to 361.7±47.9 pmol/min, and from 9.7±2.4 to 27.6±3.8 mpH/min, respectively, suggesting that energy required for activation is rapidly provided by changes in glycolytic pathways. When tested in physiological concentrations (0.1‐3 uM), malvidin and, to a certain degree, delphinidin, were more effective than cyaniding or pelargonidin in the reduction of mRNA and protein biomarkers of inflammation. Different effects exerted by individual anthocyanins suggested that the presence of the methylated hydroxyls on the B ring may be critical for their greater biological activity. Anthocyanin‐treated macrophages developed a distinct metabolic profile with respect to glycolysis versus oxidative phosphorylation, in part by increasing spare respiratory capacity and therefore shifting to mitochondria as their main source of ATP.Grant Funding Source: Supported by NCSU faculty start up funds to SK.

Synthesis of Various Esters of Diclofenac (NSAIDs) as Pro-Drugs and their Biological Evaluation

Non-steroidal anti-inflammatory drugs (NSAIDs) commonly used for the treatment of chronic inflammatory diseases suffering from several undesired side effects, the most important being gastrointestinal (GI) irritation and ulceration. The pro-drug designing is one of the several strategies used to overcome this drawback. The rationale behind the prodrug concept is to achieve temporary blockade of the free carboxylic group present in the NSAIDs till their systemic absorption. Five esters of diclofenac were synthesized with different alcohols and all the compounds were evaluated for anti-inflammatory activity by Carrageenan Induced Rat hand Paw method and surprisingly all of them showed quite appreciable anti-inflammatory activity.

A8.8 Controllable hydrogen sulfide donors and their anti-inflammatory potential in the murine macrophage cell line RAW264.7

Background and ObjectivesHydrogen sulfide (H2S) is a newly discovered gaseous signalling molecule like CO and NO and plays an important role in many physiological and/or pathological processes. Modulation of H2S...

CCR6 and CCL20: emerging players in the pathogenesis of rheumatoid arthritis

The therapeutic targeting of pro-inflammatory TNF with neutralising biological anti-TNF agents, often in combination with other disease-modifying anti-rheumatic drugs, such as the purine synthesis inhibitor methotrexate has been the first major break-through in the treatment of chronic inflammatory diseases in decades. There are however, side effects and disadvantages of these treatments, such as general immunosuppression as well as therapy resistance in a large proportion of patients. This evokes the wish for other, more specialised forms of treatments. The targeting of chemokines and their receptors to disrupt cell movement specifically has been seen as a promising avenue of therapy for a considerable time. We will discuss one particular chemokine and its receptor, the C-C chemokine ligand CCL20 and the C-C chemokine receptor CCR6, and summarise its genetic and biological role in rheumatoid arthritis (RA). CCR6 has been associated with RA in genome-wide association studies and has been shown to be an interesting candidate for a therapeutic approach, considering its and CCL20's expression patterns within the tissue as well as the immune system.

Chemical constituents in n-butanol fractions of Castus afer ker Gawl leaf and stem.

Aim:This study was designed to investigate the bioactive compounds in Costus afer Ker Gawl, an indigenous African medicinal plant whose leaf and stem extracts are used in the treatment of chronic inflammatory diseases, especially rheumatism and arthritis.Materials and Methods:The bioactive compounds present in the n-butanol fractions of C. afer leaf and stem were identified using qualitative phytochemical evaluation and gas chromatography-mass spectrometry (GC/MS) analytical method, comparing the mass spectra of the identified compounds with those of the National Institute of Standards and Technology database library.Results:Qualitative analysis detected alkaloids, saponins, diterpenes, triterpenes, phytosterol, phlobatannins, and tannins in both n-butanol fractions of C. afer leaf and stem. Phenols were detected in leaves alone while flavonoids were present in stem alone. GC/MS data showed that the bioactive compounds in n-butanol fraction of C. afer leaf were indolizine, 2-methoxy-4 vinylphenol, phytol, hexadecanoic acid-methyl ester, n-hexadecanoic acid, 9,12-octadecanoic acid-methyl ester, eicosane, cis-vaccenic acid and oleic acid while n-butanol fraction of C. afer stem contain benzofuran,2,3-dihydro,2-methoxy-4 vinylphenol, 9-octadecenoic acid (Z)-2-hydroxy-1-(hydroxymethyl) ethyl ester, campesterol, stigmasterol, hexadecanoic acid-methyl ester, n-hexadecanoic acid, and cis-vaccenic acid.Conclusion:The bioactive compounds identified in the n-butanol fractions of C. afer leaves and stem may explain the folkloric use of C. afer plant in the treatment of chronic inflammatory and oxidative stress related diseases.

Peripheral injection of SB203580 inhibits the inflammatory-dependent synthesis of proinflammatory cytokines in the hypothalamus.

The study was designed to determine the effects of peripheral injection of SB203580 on the synthesis of interleukin- (IL-) 1β, IL-6, and tumor necrosis factor (TNF) α in the hypothalamus of ewes during prolonged inflammation. Inflammation was induced by the administration of lipopolysaccharide (LPS) (400 ng/kg) over 7 days. SB203580 is a selective ATP-competitive inhibitor of the p38 mitogen-activated protein kinase (MAPK), which is involved in the regulation of proinflammatory cytokines IL-1β, IL-6 and TNFα synthesis. Intravenous injection of SB203580 successfully inhibited (P < 0.01) synthesis of IL-1β and reduced (P < 0.01) the production of IL-6 in the hypothalamus. The p38 MAPK inhibitor decreased (P < 0.01) gene expression of TNFα but its effect was not observed at the level of TNFα protein synthesis. SB203580 also reduced (P < 0.01) LPS-stimulated IL-1 receptor type 1 gene expression. The conclusion that inhibition of p38 MAPK blocks LPS-induced proinflammatory cytokine synthesis seems to initiate new perspectives in the treatment of chronic inflammatory diseases also within the central nervous system. However, potential proinflammatory effects of SB203580 treatment suggest that all therapies using p38 MAPK inhibitors should be introduced very carefully with analysis of all expected and unexpected consequences of treatment.

Anti-inflammatory effects and molecular mechanisms of loquat (Eriobotrya japonica) tea

Loquat (Eriobotrya japonica) leaves have been used as traditional medicine for chemoprevention and treatment of chronic inflammatory diseases although the evidence supporting their functions is still poor. This study aims to clarify the anti-inflammatory effects and molecular mechanisms of loquat tea extract (LTE), in both cell and animal models. LTE, especially C fraction, inhibited the production of pro-inflammatory factors including inducible nitric oxide synthase (iNOS), nitric oxide (NO), IL-6, RANTES and TNF-α. Cellular signaling data revealed that the downregulation of TGF-β-activated kinase (TAK1)-mediated both mitogen-activated protein kinase (MAPK) and NF-κB pathways were involved in the inhibition of pro-inflammatory factors by C fraction. Mouse paws edema model further confirmed the in vivo anti-inflammatory effects of LTE.

Pharmacophore Modeling and Virtual Screening for the Discovery of New type 4 cAMP Phosphodiesterase (PDE4) Inhibitors

Type 4 cAMP phosphodiesterase (PDE4) inhibitors show a broad spectrum of anti-inflammatory effects in almost all kinds of inflamed cells, by an increase in cAMP levels which is a pivotal second messenger responsible for various biological processes. These inhibitors are now considered as the potential drugs for treatment of chronic inflammatory diseases. However, some recently marketed inhibitors e.g., roflumilast, have shown adverse effects such as nausea and emesis, thus restricting its use. In order to identify novel PDE4 inhibitors with improved therapeutic indexes, a highly correlating (r = 0.963930) pharmacophore model (Hypo1) was established on the basis of known PDE4 inhibitors. Validated Hypo1 was used in database screening to identify chemical with required pharmacophoric features. These compounds are further screened by using the rule of five, ADMET and molecular docking. Finally, twelve hits which showed good results with respect to following properties such as estimated activity, calculated drug-like properties and scores were proposed as potential leads to inhibit the PDE4 activity. Therefore, this study will not only assist in the development of new potent hits for PDE4 inhibitors, but also give a better understanding of their interaction with PDE4. On a wider scope, this will be helpful for the rational design of novel potent enzyme inhibitors.