Increased gene expression of inflammatory markers in nasal turbinate of patients with persistent allergic rhinitis and chronic obstruction.

PURPOSE: The pathogenesis of persistent allergic rhinitis with chronic and refractory nasal obstruction is still unknown. Inflammation and tissue remodeling are known to play a role, but this has not been studied thoroughly. The purpose of this study is to identify the profile of gene expression of inflammatory and remodeling markers in nasal mucosa of patients with PAR and chronic obstruction. METHODS: After informed consent, we obtained nasal mucosa tissue from five aeroallergen-sensitized PAR patients undergoing anterior turbinectomy, and control non-sensitized individuals undergoing cerebrospinal fluid fistula repair or rhinoplasty. We assessed the expression of 34 genes related to inflammation and tissue remodeling using the real-time polymerase chain reaction (qPCR) to quantify each mRNA. RESULTS: IL-4 mRNA was upregulated in nasal mucosa of all five patients; CCR3, CCR8 and Eotaxin-2 were upregulated in four out of five patient samples; while IL-5 and IL-13 were upregulated in two of them. TGF-ß1 was not upregulated in PAR samples. mRNA from metalloproteinases MMP-7, MMP13 and MMP15 were upregulated in three out of five samples. Our results indicate a typical mRNA expression profile of the infiltrating inflammatory Th2 cells and eosinophils, combined with altered gene expression of remodeling-related proteins in stromal cells from the mucosa. CONCLUSION: Prolonged allergen challenge can lead to persistent upregulation of genes for inflammatory mediators such as IL-4 Th2/eosinophil cytokines, chemokines and receptors, which may play an important role in maintaining PAR with chronic nasal obstruction. Our findings may have therapeutic implications, including the use of anti-IL4, -CCR3 or -MMP therapy to ameliorate the condition.

Efeitos clínicos e imunológicos de ácidos graxos poli-insaturados de ômega-3 e aspirina em baixa dosagem como adjuvantes ao debridamento periodontal em pacientes com diabetes tipo 2: estudo clínico randomizado / Clinical and immunological effects of omega-3 polyunsaturated fatty acids and low-dose aspirin as adjuncts to periodontal debridement in patients with type 2 diabetes: randomized clinical trial

A suplementação diária com ácidos graxos poli-insaturados de ômega-3 (ω-3) e a aspirina em baixa dosagem foram propostas como terapia de modulação do hospedeiro para o tratamento de doenças inflamatórias crônicas. O objetivo deste estudo foi investigar as ações clínicas e imunológicas do ω-3 e da aspirina (AAS) como terapia adjunta ao debridamento periodontal de boca toda para o tratamento da periodontite em pacientes com diabetes tipo 2. Setenta e cinco pacientes (n=25/grupo) que atendiam aos critérios de inclusão foram randomicamente designados para receber placebo e debridamento periodontal (GC), ω-3 (3g de óleo de peixe/dia por 60 dias) e AAS (100mg/dia por 60 dias) após o debridamento periodontal (GT1), e (3g de óleo de peixe/dia por 60 dias) e AAS (100mg/dia por 60 dias) antes do debridamento periodontal (GT2). Parâmetros clínicos periodontais e fluido gengival crevicular (FGC) foram coletados no baseline (t0), 90 dias (GT1 e GC) (t1), após a suplementação/medicação com ω-3 e AAS (t1), e 180 dias após o debridamento periodontal (todos os grupos) (t2). Dez pacientes (40%) no GT1 e nove pacientes (36%) no GT2 alcançaram o endpoint clínico para o tratamento (≤4 bolsas periodontais com profundidade de sondagem (PS)≥ 5mm), em contraste com quatro (16%) no GC. Houve ganho de inserção em bolsas moderadas e em bolsas profundas entre t0 e t2 para o GT1. Os níveis de concentração de IFN-γ, IL-1ß e IL-8 apresentaram redução em t2 para os dois grupos teste, com mudanças significantes prévias (t1) para o GT1. Os níveis de IL-6 apresentaram redução em t1 e em t2 para o GT1, e a MIP-1α reduziu em t2 no GT2. No GC a IL-1ß foi a única citocina a apresentar diferença estatisticamente significante na comparação entre tempos. Os resultados deste estudo clínico sugerem que a terapia adjuvante de ω-3 a AAS após o debridamento periodontal promove maiores benefícios clínicos e imunológicos ao tratamento da periodontite em pacientes com diabetes tipo 2 quando comparado aos demais protocolos avaliados(AU)

Daily dietary supplementation with omega-3 (ω-3) polyunsaturated fatty acids (PUFAs) and low-dose aspirin (ASA) have been proposed as a host modulation therapy for the treatment of chronic inflammatory diseases. The aim of this study was to investigate the clinical and immunological actions of ω-3 PUFAs and ASA as an adjunct therapy to full-mouth periodontal debridement for the treatment of periodontitis in patients with type 2 diabetes. Seventy-five patients (n=25/group) meeting the inclusion criteria were randomly assigned to receive placebo and periodontal debridement (CG), ω-3 (3g of fish oil/day for 60 days) and ASA (100mg/day for 60 days) after periodontal debridement (TG1), and ω-3 (3g of fish oil/day for 60 days) and ASA (100mg/day for 60 days) before periodontal debridement (TG2). Periodontal clinical parameters and gingival crevicular fluid (GCF) were collected at baseline (t0), 90 days (TG1 and CG) (t1), after ω-3 and ASA only (TG2) (t1), and 180 days after periodontal debridement (all groups) (t2). Ten patients (40%) in TG1 and nine patients (36%) in TG2 achieved the clinical endpoint for treatment (≤4 periodontal pockets with probing depth (PD)≥ 5mm), as opposed to four (16%) in CG. There was clinical attachment gain in moderate and deep pockets between t0 and t2 for TG1. Concentration levels of IFN-γ, IL-1ß, and IL-8 decreased over time for both test groups, with early (t1) significant changes for TG1. IL-6 levels were lower at t1 and t2 for TG1, and MIP-1α decreased at t2 for TG2. In the CG, IL1ß was the only marker presenting statistically significant changes over time. The results of this clinical study suggest that the adjunctive use of ω-3 and ASA after periodontal debridement provides clinical and immunological benefits to the treatment of periodontitis in patients with type 2 diabetes when compared to the other treatment protocols evaluated(AU)

The influence of cardiac resynchronization therapy on selected inflammatory markers and aldosterone levels in patients with chronic heart failure.

The aim of the study was to assess the influence of cardiac resynchronization therapy(CRT) on a series of humoral parameters crucial for the pathophysiology of chronic heart failure such as aldosterone or the inflammatory markers. Thirty eight consecutive patients (aged 66.3 +/- 9.6 years, 31 men - 82% ) with chronic heart failure (57.9% with ischaemic background and 42.1% of non-ischaemic etiology) in stable for at least 3 months, NYHA class III - IV despite optimized pharmacotherapy, with left ventricular ejection fraction (LVEF) < or = 35% and wide QRS complex (> or = 120 ms) had the blood serum tested for the concentrations of interleukin-6 (IL-6), interleukin-18 (IL-18), C-reactive protein (CRP) and aldosterone before and 12-16 weeks after CRT introduction. In the study group aldosterone concentrations were significantly reduced. Among the inflammatory markers the level of IL-6 decreased, IL-18 concentrations showed a falling trend (445.1 +/- 225.7 pg/ml vs 418.4 +/- 229.6 pg/ml, p = 0.052), whereas no change of CRP serum contain was noted. It was showed that cardiac resynchronization therapy had an impact on systemic inflammation and hormonal status in patients with chronic heart failure during short-term observation.

IL-1α and IL-1ß recruit different myeloid cells and promote different stages of sterile inflammation.

The immune system has evolved to protect the host from invading pathogens and to maintain tissue homeostasis. Although the inflammatory process involving pathogens is well documented, the intrinsic compounds that initiate sterile inflammation and how its progression is mediated are still not clear. Because tissue injury is usually associated with ischemia and the accompanied hypoxia, the microenvironment of various pathologies involves anaerobic metabolites and products of necrotic cells. In the current study, we assessed in a comparative manner the role of IL-1α and IL-1ß in the initiation and propagation of sterile inflammation induced by products of hypoxic cells. We found that following hypoxia, the precursor form of IL-1α, and not IL-1ß, is upregulated and subsequently released from dying cells. Using an inflammation-monitoring system consisting of Matrigel mixed with supernatants of hypoxic cells, we noted accumulation of IL-1α in the initial phase, which correlated with the infiltration of neutrophils, and the expression of IL-1ß correlated with later migration of macrophages. In addition, we were able to show that IL-1 molecules from cells transfected with either precursor IL-1α or mature IL-1ß can recruit neutrophils or macrophages, respectively. Taken together, these data suggest that IL-1α, released from dying cells, initiates sterile inflammation by inducing recruitment of neutrophils, whereas IL-1ß promotes the recruitment and retention of macrophages. Overall, our data provide new insight into the biology of IL-1 molecules as well as on the regulation of sterile inflammation.

The immune effects of naturally occurring and synthetic nanoparticles.

Ultrafine particles and engineered nanoparticles have unique aerodynamic and biochemical properties that affect the immune system and human health in ways that are different from or exceed those seen with gases or larger particulates. These effects result from a unique set of physical characteristics and surface moieties, which generate an ability of UFPs to enter tissues and cells, interact with proteins and DNA at a molecular level and directly and indirectly modulate the immune system by novel mechanisms. In recent years, a new field known as nanotechnology has impacted multiple industries by taking advantage of the special qualities of these small "atomic-sized" particles. Nanomedicine has already opened up a new avenue of research in cancer therapy, drug delivery and immune regulation. While the benefits of this new science to human civilization are seemingly immeasurable, it is also important to appreciate that these particles can also lead to harmful effects on human health. In vitro and animal studies are showing that nanoparticles and UFPs are capable of activating proinflammatory cytokines, chemokines and adhesion molecules, with recruitment of inflammatory cells including basophils, macrophages, dendritic cells, T cells, neutrophils and eosinophils. These changes may have an impact on immune defense, but also on the Th1/Th2 balance, and even on non-immunologic function. Resulting immune system derangement can lead to increases in incidence of autoimmune, allergic and even neoplastic diseases. Cardiorespiratory effects have been observed to occur in humans. Much further research is needed to establish safe exposure levels for this important new class of particulates.

Alum induces innate immune responses through macrophage and mast cell sensors, but these sensors are not required for alum to act as an adjuvant for specific immunity.

To understand more about how the body recognizes alum we characterized the early innate and adaptive responses in mice injected with the adjuvant. Within hours of exposure, alum induces a type 2 innate response characterized by an influx of eosinophils, monocytes, neutrophils, DCs, NK cells and NKT cells. In addition, at least 13 cytokines and chemokines are produced within 4 h of injection including IL-1beta and IL-5. Optimal production of some of these, including IL-1beta, depends upon both macrophages and mast cells, whereas production of others, such as IL-5, depends on mast cells only, suggesting that both of these cell types can detect alum. Alum induces eosinophil accumulation partly through the production of mast cell derived IL-5 and histamine. Alum greatly enhances priming of endogenous CD4 and CD8 T cells independently of mast cells, macrophages, and of eosinophils. In addition, Ab levels and Th2 bias was similar in the absence of these cells. We found that the inflammation induced by alum was unchanged in caspase-1-deficient mice, which cannot produce IL-1beta. Furthermore, endogenous CD4 and CD8 T cell responses, Ab responses and the Th2 bias were also not impacted by the absence of caspase-1 or NLRP3. These data suggest that activation of the inflammasome and the type 2 innate response orchestrated by macrophages and mast cells in vivo are not required for adjuvant effect of alum on endogenous T and B cell responses.

Synergistic drug-cytokine induction of hepatocellular death as an in vitro approach for the study of inflammation-associated idiosyncratic drug hepatotoxicity.

Idiosyncratic drug hepatotoxicity represents a major problem in drug development due to inadequacy of current preclinical screening assays, but recently established rodent models utilizing bacterial LPS co-administration to induce an inflammatory background have successfully reproduced idiosyncratic hepatotoxicity signatures for certain drugs. However, the low-throughput nature of these models renders them problematic for employment as preclinical screening assays. Here, we present an analogous, but high-throughput, in vitro approach in which drugs are administered to a variety of cell types (primary human and rat hepatocytes and the human HepG2 cell line) across a landscape of inflammatory contexts containing LPS and cytokines TNF, IFN gamma, IL-1 alpha, and IL-6. Using this assay, we observed drug-cytokine hepatotoxicity synergies for multiple idiosyncratic hepatotoxicants (ranitidine, trovafloxacin, nefazodone, nimesulide, clarithromycin, and telithromycin) but not for their corresponding non-toxic control compounds (famotidine, levofloxacin, buspirone, and aspirin). A larger compendium of drug-cytokine mix hepatotoxicity data demonstrated that hepatotoxicity synergies were largely potentiated by TNF, IL-1 alpha, and LPS within the context of multi-cytokine mixes. Then, we screened 90 drugs for cytokine synergy in human hepatocytes and found that a significantly larger fraction of the idiosyncratic hepatotoxicants (19%) synergized with a single cytokine mix than did the non-hepatotoxic drugs (3%). Finally, we used an information theoretic approach to ascertain especially informative subsets of cytokine treatments for most highly effective construction of regression models for drug- and cytokine mix-induced hepatotoxicities across these cell systems. Our results suggest that this drug-cytokine co-treatment approach could provide a useful preclinical tool for investigating inflammation-associated idiosyncratic drug hepatotoxicity.

New insights into the evolution of IFNs: zebrafish group II IFNs induce a rapid and transient expression of IFN-dependent genes and display powerful antiviral activities.

The IFNs and their receptors have existed in early chordates for approximately 500 million years and represent the early elements in innate and adaptive immunity. Both types I and II IFNs have been discovered in fish, and type I has recently been classified into two groups based on their primary protein sequences. However, the biological activities of fish IFNs and their roles in infection are largely unknown. Using the zebrafish and manageable bacterial (Streptococcus iniae) and viral (spring viremia of carp virus) infection models, we are reporting in this study that zebrafish IFN (zfIFN) gamma failed to induce antiviral and proinflammatory genes when administered in vivo, which correlates with its inability to protect the fish against bacterial and viral infections. We also found that, although both group I (i.e., zfIFN1) and group II zfIFNs (i.e., zfIFN2 and zfIFN3) displayed strong in vivo antiviral activities, only group I zfIFN was able to protect the fish against bacterial infection, which may reflect the different patterns and kinetics of immune-related genes elicited by these two groups of IFNs. Thus, group II zfIFNs induced a rapid and transient expression of antiviral genes, whereas group I zfIFN exerted a slow but more powerful induction of several antiviral and proinflammatory genes. Collectively, our results suggest nonredundant, complementary roles of type I zfIFNs in viral infections and provide evidence for a pivotal role of the recently identified group II IFN of fish in the early stages of viral infections.

The role of serpins in vertebrate immunity.

Serine proteases are important components of the immune system, playing a role in many processes including migration, phagocytosis and elimination of virally infected and cancerous cells. Members of the serpin superfamily regulate the activity of these proteases to limit tissue damage and unwarranted cell death. This review focuses on the role of intracellular (clade B) serpins in maintaining viability of both innate and adaptive immune cells.

Inflammatory markers in stable heart failure and their relationship with functional class.

INTRODUCTION AND OBJECTIVES: While it appears to be clear that an inflammatory process occurs in heart failure (HF), it is still to be defined whether inflammation depends to a greater extent on HF etiology, functional class (FC), or the extent of depression of ejection fraction (EF). Our objectives were to analyze differences in inflammatory marker levels as compared to a healthy population, to assess differences depending on HF etiology, and to relate values with FC and EF. PATIENTS AND METHODS: Fifty-nine consecutive outpatients with stable HF (57 + or - 9 years, 89% males) and 59 controls (55 + or - 8 years, 85% males) were enrolled into the study. Causes of HF included ischemic heart disease (n=24), idiopathic dilated cardiomyopathy (n=24), and miscellaneous conditions (n=11). Patients with decompensation in the past 6 months were excluded from the study. Protein fibrinogen, sialic acid, C-reactive protein (CRP), and tumor necrosis factor-alpha (TNF-alpha) were measured. Echocardiography was performed in all study patients. FC was assessed using the NYHA classification. RESULTS: A comparison of inflammatory marker levels between the HF and control groups showed significant differences in all markers, except for TNF-alpha. Protein fibrinogen in controls: 253 + or - 54 mg/dl, protein fibrinogen in HF: 294 + or - 67 mg/dl; p<0.05. Sialic acid in controls: 53 + or - 1 mg/dl, sialic acid in HF: 61 + or - 12 mg/dl; p<0.05. CRP in controls: 1.3 + or - 0.7 mg/dl, CRP in HF: 7.8 + or - 1.2 mg/dl; p<0.05. TNF-alpha in controls: 183 + or - 51 ng/ml, TNF-alpha in HF: 203 + or - 13 ng/ml; p=0.2. No differences were found between the different etiologies of HF. A positive association was seen between FC and protein fibrinogen and TNF-alpha (p<0.05), but not with EF. CONCLUSIONS: Increased inflammatory marker levels related to FC of the patient, but not to EF, are found in chronic HF.

Fine urban atmospheric particulate matter modulates inflammatory gene and protein expression in human bronchial epithelial cells.

Ambient particulate matter (PM) is known to induce inflammation in the respiratory tract of exposed subjects. The aim of the present study was to detect, in bronchial epithelial cells, candidate inflammatory genes exhibiting transcriptional modifications following urban PM2.5 exposure. Paris urban PM2.5 sampled either at a curbside or a background station in winter and in summer was tested in comparison with diesel exhaust particles (DEP) at 10 microg/cm2 on human bronchial epithelial (16-HBE) cells (18 h of exposure). The gene profiling study performed using a 375 cDNA cytokine expression array highlighted the differential expression of certain genes, three of which were selected as genes of interest: the IL-1 alpha cytokine, the GRO-alpha chemokine, and amphiregulin, a ligand of the EGF receptor. Their increased expression was confirmed by RT-PCR and/or by Northern blotting in bronchial epithelial cells. In the culture medium of particle-treated cultures, increased release of GRO-alpha and amphiregulin was shown. The particle component responsible for protein release varied for the two genes. The organic extract seemed to be mainly involved in amphiregulin expression and secretion, whereas both the aqueous and organic extracts induced GRO-alpha release. In conclusion, in bronchial epithelial cells, Paris PM2.5 increased mRNA and protein expression of GRO-alpha and AR involved in the chemoattraction process and bronchial remodeling, respectively.

Mediators of chronic obstructive pulmonary disease.

Chronic obstructive pulmonary disease (COPD) is a major and increasing global health problem that is now a leading cause of death. COPD is associated with a chronic inflammatory response, predominantly in small airways and lung parenchyma, which is characterized by increased numbers of macrophages, neutrophils, and T lymphocytes. The inflammatory mediators involved in COPD have not been clearly defined, in contrast to asthma, but it is now apparent that many lipid mediators, inflammatory peptides, reactive oxygen and nitrogen species, chemokines, cytokines, and growth factors are involved in orchestrating the complex inflammatory process that results in small airway fibrosis and alveolar destruction. Many proteases are also involved in the inflammatory process and are responsible for the destruction of elastin fibers in the lung parenchyma, which is the hallmark of emphysema. The identification of inflammatory mediators and understanding their interactions is important for the development of anti-inflammatory treatments for this important disease.

Novel endogenous small molecules as the checkpoint controllers in inflammation and resolution: entrée for resoleomics.

Endogenously-generated small chemical mediators or autacoids play key roles in controlling inflammation and its organized resolution. Among them, lipoxins are the trihydroxy-tetraene-containing eicosanoids that are generated primarily by tight cell-cell interactions by way of transcellular biosynthesis and serve as local endogenous anti-inflammatory mediators. These "stop signals" in inflammation and other related processes may be involved in switching the cellular response from additional PMN recruitment toward monocytes (in a nonphlogistic fashion) that could lead to resolution of the inflammatory response or promotion of repair and healing. ASA impinges on this homeostatic system and evokes the endogenous biosynthesis of the carbon 15 epimers of lipoxins, namely ATLs, that mimic the bioactions of native LX in several biologic systems and, thus, can modulate in part, the beneficial actions of ASA in humans. Moreover, the temporal and spatial components in LX formation and actions are important determinants of their impact during an acute inflammatory reaction. Generation of lipid (ie, ATL) versus protein (ie, ANXA1) mediators during the host inflammatory response display different time courses. The temporal difference suggests that ALX could regulate PMN by interacting with each class of ligands within specific phases of the inflammatory response. ALX is the first cloned lipoxygenase-derived eicosanoid receptor. The signaling pathways and bioactions of ALX are cell type-specific. In agreement with in vitro results, ALX agonists, namely LXA4 and 15-epi-LXA4 and their stable analogs, regulate PMN during acute inflammation. In addition, it seems that LXs also display organ-specific actions, in addition to host defense and immune roles in the eye, kidney, lung, and oral and gastrointestinal tract and within bone marrow progenitors, possibly involving stem cells. The development of these few synthetic stable analogs has provided valuable tools to evaluate the biologic roles, significance, and pharmacologic actions of ALX and provided novel therapies for inflammatory diseases. The relationship between LX generation and current NSAID therapies is more intertwined than currently appreciated. ASA inhibits COX-1 and converts COX-2 into an ASA-triggered lipid mediator-generating system that produces an array of novel endogenous local autacoids from dietary omega-3 PUFA. Some of the local autacoids display potent anti-inflammatory or antineutrophil recruitment activity as well as impinge on the role of these compounds in resolution, and, thus, are termed "resolvins." It is not surprising that investigators recently found a protective action for COX-2 in cardiovascular disease. Together with the lipoxins and 15-epi-lipoxins, the identification of the resolvins gives us new avenues of approach in considering therapies for inflammation, cardiovascular diseases and cancer.

Early detection and markers of sepsis.

In vitro and animal models of sepsis have provided a template for studies of the pathogenesis of sepsis in patients at risk for and with the syndrome. Numerous potential markers have been identified in these models and then looked for in patients. No single marker or combination of markers convincingly identifies sepsis, predicts the development of sepsis, predicts the development of complications of sepsis, or predicts mortality. As discussed, the clinical studies have been complicated by many confounding variables, including the lack of adherence to rigorous definitions, differences in assay methods, differences in timing of the studies, and differences in outcome variables analyzed. In spite of the limitations, the studies have been critical in helping determine the pathogenesis of sepsis in humans. As new mediators and modulators of inflammation are identified, it will be important to study their role as markers, individually and in combination, in human disease.

Las quimoquinas : una nueva familia de citoquinas reguladoras del proceso inflamatorio

La respuesta inmune e inflamatoria es regulada positiva y negativamente por medio de la secreción de mediadores solubles denominados citoquinas. Una nueva familia de citoquinas denominadas quimoquinas se compone por lo menos de 18 proteínas. Estas citoquinas presentan 20 a 45 por ciento de homología entre su secuencia de aminoácidos y poseen cuatro cisteínas conservadas que forman enlaces disulfuro en el extremo NH2 de la proteína. Con base en la secuencia de las primeras dos cisteínas, las quimoquinas se han dividido en dos subfamilias; las B-quimoquinas en las cuales las primeras dos cisteínas se encuentran separadas por un residuo no conservado (C-X-C) y las B-quimoquinas en las cuales estas dos cisteínas se encuentran adyacentes (C-C). Dentro de las actividades biológicas que comparten las alfa y las beta-quimoquinas se encuentran el incremento pasajero del calcio intracelular, cambios de la forma celular al interactuar con proteínas de la matriz celular, inducción de degranulación, incremento en la expresión de moléculas de adhesión celular (B2 integrinas), producción de prostaglandinas y leucotrienos e inducción de la explosión respiratoria en neutrófilos, basófilos, linfocitos T y monocitos principalmente. De manera particular, las B-quimoquinas ejercen una modulación de la respuesta inmunológica a través de varias actividades pleitrópicas, las cuales incluyen el reclutamiento de Células T y macrófagos en sitios de inflamación por medio de la regulación de su activación y proliferación. Se ha desmostrado recientemente que estas funciones pleiotrópicas de las quimoquinas juegan un papel importante en la regulación del proceso inflamatorio asociado al desarrollo de la patología de procesos autoinmunes y alérgicos.