Recruitment Of Mononuclear Cells Research Articles

Haplotype Analysis of TNFA Gene in Peptic Ulcer Patients

The TNFA gene product TNF- is a cytokine promoting an immune response after bacterial infections. An excessive production of the cytokine is thought to be connected with Helicobacter pylori-induced disorders like gastritis, peptic ulcer disease (PUD), intestinal metaplasia, or even gastric cancer. The synthesis of TNF-a is controlled at transcriptional level and dependent on single nucleotide polymorphisms (SNPs) of TNFA promoter region. SNPs assembled in haplotype have been implicated as potential risk factors for various autoimmune and infectious diseases. The aim of this study was to determine the frequencies of TNFA haplotypes composed of the four common single-nucleotide polymorphisms of the gene (-1031C>T: rs1799964, -863C>A: rs1800630, 857C>T: rs1799724, -308G>A: rs1800629) in peptic ulcer patients and to assess the significance of the haplotypes as the risk factors of H. pylori infection development. 203 peptic ulcer patients were genotyped using polymerase chain reaction-restriction fragment length polymorphism method. Haplotypes and degree of linkage disequilibrium (LD) were inferred with PHASE 2.1 and EMLD software. There was no statistically significant difference in haplotype frequencies between the H. pylori-infected and -uninfected peptic ulcer cases (p=0.62). Analogous association was also absent in the subgroups: peptic ulcer woman (p=0.69), peptic ulcer men (p=0.17). The locus pairs -308_-857, -863_-1031, and -857_-1031 was found to be in very strong LD , -308_-1031 and -857 and -863 in strong LD, and -308_-863 in modest LD. TNFA haplotype structure is not connected with individual differences in susceptibility to development of H. pylori infection in peptic ulcer patients. Address for correspondence: Dr. Aleksandra Salagacka Medical University of Lodz Musuynskiego 1, 90-151, Lodz, Poland Telephone/Fax: +48 42 677 91 30 E-mail: aleksandra.salagacka@umed.lodz.pl INTRODUCTION Tumour necrosis factor-alpha (TNF-) is a cytokine known for having the multitude of functions. One of the most prominent is to promote of immune response after bacterial infections. In gastric mucosa infected by Helicobacter pylori TNF- prompts recruitment of mononuclear cells and neutrophils into the gastric lamina propria (Crabtree 1991; Huseyinov 1999). An excessive production of the cytokine could also exert some pathogenic effects (Beutler 1993) and is thought to be connected with H. pylori-induced disorders like gastritis, peptic ulcer disease (PUD), intestinal metaplasia, or even gastric cancer (Goto 2003; Shanks 2009). The synthesis of TNF- is controlled at transcriptional level in a cell typeand stimulus-specific manner involving different transcription factors to a promoter region of the TNFA encoding TNF- (Falvo 2010). An important role in the regulation of TNFA expression is attributed to single nucleotide polymorphisms (SNPs) of promoter region of the gene. Accordingly, these genetic variations are implicated as potential risk factors for various autoimmune and infectious diseases. To date, SNPs at positions -238, -308, -857, 863, -1031 of TNFA have been investigated in various populations afflicted by PUD or other H. pylori-induced disorders (Lee 2005; Lu 2005; Lee 2004; Wilschanski 2007; Kim 2006; Machado 2003). Unfortunately, studies focused on determining the importance of individual TNFA loci have produced non-significant or conflicting results. However, some evidence has supported the hypothesis that effects exerts by these SNPs could be observed or are more pronounced when the polymorphisms are researched and analysed concomitantly. Studies by Lu et al. (2005) conducted in Taiwanese revealed that risk of ulceration after H. pylori infection was higher for carriers of both -1031C or -863A allele of TNFA than for individuals having only one of the mentioned alleles. Similarly, the risk of gastric ulcers among Japanese was found to be greatest in simultaneous carriers of so called ‘high-producer’ alleles of TNFA -857/-863/-1031 than when the presence of these particular alleles was stated individually (Sugimoto 2007). 10 ALEKSANDRA SALAGACKA, MARTA ZEBROWSKA, AGNIESZKA JELEN ET AL Some research results could imply the best approach to investigate the relevancy of TNFA variations, in the development of gastroduodenal diseases, is to explore haplotype structure of TNFA gene. Although Chakravorty et al. (2008) did not stated any connection between single TNFA loci (-308, -857, -863, -1031) and H. pylorimediated duodenal ulcer, the researchers found the TNFA haplotype -308G_-857C_-863A_-1031T in H. pylori-infected duodenal ulcer individuals was significantly more frequent than in the infected but ulcer-free cases. Thus far, haplotype analysis of the gene has been applied in the studies on type 1 diabetes (Stayoussef 2010), gastric cancer (Canedo 2008), iron deficiency anemia (Atkinson 2008), Behcet’s disease (Park 2006), schizophrenia (Saviouk 2005), juvenile oligoarthritis (Zeggini 2002), hepatitis C virus infection (Thio 2004), and some others. Recently, the researchers investigated association between individual -308G>A or -1031T>C SNPs of TNFA and peptic ulcer risk in Poles (Salagacka 2014). In this study the researchers conducted haplotype analysis of TNFA combining mentioned genotyping data and newly obtained for -308 locus and two further -857 and -863 loci of TNFA. The aim of presented research was to estimate the TNFA haplotype structure and its significance in development peptic ulcer disease in Polish population.

Uric Acid and Coronary Collateral Circulation

Coronary collateral (CC) artery development is usually attributed to ischemia, but these anastomotic channels are also present in individuals who do not have coronary artery disease (CAD). The CCs are present in one-fourth of the patients with normal coronary arteries or nonobstructive CAD. Currently, there is no noninvasive method for evaluating CC circulation in humans. The most widely used strategy is visual assessment of collateral arteries as described by Rentrop et al. This method involves balloon occlusion of the contralateral coronary artery, which is rarely performed. In clinical practice, this method is applied without occluding the contralateral vessels. As a result, underestimation of collateralization is inevitable. Other limitations of this visual method include observer variability, influence by the blood pressure, and the force of contrast injection as well as the duration of filming. The most accurate assessment could be obtained by measuring the pressure-derived collateral flow index. Several clinical and angiographic variables including the degree of coronary stenosis, proximal lesion location, longer duration of symptoms, and occlusion as well as resting bradycardia have been reported as variables that correlated with the degree of collateralization. Also, in patients without CAD, the baseline heart rate has been described as the main predictor of collateralization. The most important trigger for collateral growth is tangential (radial) fluid shear stress at the endothelial level with the recruitment of bone marrow-derived mononuclear cells. It is a remodeling process of preexisting small collateral arterioles rather than the growth of new capillary vessels, which is induced by ischemia. Collateral growth is induced by fluid shear stress in preformed collateral vessels caused by a pressure gradient between the area proximal to a coronary stenosis and the low-pressure poststenotic area. The CCs play a role in preserving myocardial function, limiting infarct size, and increasing survival. However, a meta-analysis showed that a well-developed collateral circulation was a risk factor for restenosis after coronary revascularization. In humans, uric acid (UA) is generated from purine catabolism with the conversion of hypoxanthine to xanthine and of xanthine to UA by the enzyme xanthine oxidoreductase. It is further oxidized to allantoin in most species except humans and some higher primates who lack uricase. Therefore, serum UA (SUA) levels in humans are appreciably higher than in other mammals. Because of this difference in urate metabolism, the results of animal studies can be difficult to interpret. Also, different methods of measuring collateral flow may be applied in animal studies. Many patients with CAD use at least 1 drug that can influence SUA levels (eg, aspirin, statins, fibrates, and some antihypertensive agents). The SUA levels are higher in men than in women at all ages due to gender steroids. Sumino et al showed that hormone replacement therapy in postmenopausal women was associated with a significant reduction in SUA level. Also, dietary, environmental, and genetic factors are important determinants of SUA levels. It is important to note that UA is a potent antioxidant, but it can also be a prooxidant if the level is >6 mg/dL in women and 6.5 to 7.0 mg/dL in men. Therefore, elevated SUA level might be an indicator of increased burden of oxidant attack. Besides its dual oxidant effect, if UA is a harmful product, why do our kidneys recover 90% of filtered UA instead of eliminating it? Despite the antioxidant properties of UA, several studies have indicated that high SUA levels are associated with several disorders, including CAD, peripheral arterial disease, heart failure, metabolic syndrome, hypertension, and stroke. These associations have been mainly attributed to upregulation of renin release and the subsequent cascade-related reduction in endothelial function. However, instead of proven causal associations, possible explanations with small study groups could be reverse causality. For example, preclinical atherosclerosis could lead to higher levels of SUA. In contrast, experts from the Framingham Heart Study group have reported no association between the SUA and the cardiovascular disease. In a recently published study, Palmer et al investigated the association of plasma UA with CAD and hypertension using Mendelian randomization. They found no evidence of a causal effect of UA or hyperuricemia on the risk of CAD and hypertension. Recently, some studies have demonstrated an association between SUA and CC development in patients with CAD and

Endothelial Junctional Adhesion Molecule-A Guides Monocytes Into Flow-Dependent Predilection Sites of Atherosclerosis

Junctional adhesion molecule (JAM)-A expressed in endothelial, epithelial, and blood cells can regulate permeability and leukocyte extravasation. Atherosclerosis develops at sites of disturbed flow in large arteries, but the mechanisms guiding inflammatory cells into these predilection sites remain unknown. To characterize cell-specific functions of JAM-A in atherosclerosis, we used apolipoprotein E-deficient mice with a somatic or endothelium-specific deficiency in JAM-A and bone marrow chimeras with JAM-A-deficient leukocytes. We show that impaired JAM-A expression in endothelial cells reduced mononuclear cell recruitment into the arterial wall and limited atherosclerotic lesion formation in hyperlipidemic mice. In contrast, JAM-A deficiency in bone marrow cells impeded monocyte de-adhesion, thereby increasing vascular permeability and lesion formation, whereas somatic JAM-A deletion revealed no significant effects. Regions with disturbed flow displayed a focal enrichment and luminal redistribution of endothelial JAM-A and were preferentially protected by its deficiency. The functional expression and redistribution of endothelial JAM-A was increased by oxidized low-density lipoprotein, but confined by atheroprotective laminar flow through an upregulation of microRNA (miR)-145, which repressed JAM-A. Our data identify endothelial JAM-A as an important effector molecule integrating atherogenic conditions to direct inflammatory cell entry at predilection sites of atherosclerosis.

Inhibition of Notch signaling induces extensive intussusceptive neo-angiogenesis by recruitment of mononuclear cells

Notch is an intercellular signaling pathway related mainly to sprouting neo-angiogenesis. The objective of our study was to evaluate the angiogenic mechanisms involved in the vascular augmentation (sprouting/intussusception) after Notch inhibition within perfused vascular beds using the chick area vasculosa and MxCreNotch1(lox/lox) mice. In vivo monitoring combined with morphological investigations demonstrated that inhibition of Notch signaling within perfused vascular beds remarkably induced intussusceptive angiogenesis (IA) with resultant dense immature capillary plexuses. The latter were characterized by 40 % increase in vascular density, pericyte detachment, enhanced vessel permeability, as well as recruitment and extravasation of mononuclear cells into the incipient transluminal pillars (quintessence of IA). Combination of Notch inhibition with injection of bone marrow-derived mononuclear cells dramatically enhanced IA with 80 % increase in vascular density and pillar number augmentation by 420 %. Additionally, there was down-regulation of ephrinB2 mRNA levels consequent to Notch inhibition. Inhibition of ephrinB2 or EphB4 signaling induced some pericyte detachment and resulted in up-regulation of VEGFRs but with neither an angiogenic response nor recruitment of mononuclear cells. Notably, Tie-2 receptor was down-regulated, and the chemotactic factors SDF-1/CXCR4 were up-regulated only due to the Notch inhibition. Disruption of Notch signaling at the fronts of developing vessels generally results in massive sprouting. On the contrary, in the already existing vascular beds, down-regulation of Notch signaling triggered rapid augmentation of the vasculature predominantly by IA. Notch inhibition disturbed vessel stability and led to pericyte detachment followed by extravasation of mononuclear cells. The mononuclear cells contributed to formation of transluminal pillars with sustained IA resulting in a dense vascular plexus without concomitant vascular remodeling and maturation.

Elastase-coupled beads as a tool for characterizing localized alveolar tissue destruction associated with the onset of emphysema.

Intratracheal elastase challenge of laboratory animals has long been established as a model for observing the physiological and morphological changes that result from alveolar destruction, the hallmark of emphysema. However, instillation of elastase suspended in buffer results in widespread inflammation and variable emphysematous lesions, which has made the identification of specific cellular and molecular events associated with the onset of emphysema difficult to define. Here we establish a bead-based elastase delivery system that induces localized tissue destruction, a key event in the initiation of emphysema. Elastase was coupled to bisacrylamide beads, which were shown to retain enzymatic activity prior to intratracheal administration in mice. C57BL/6 mice were given a single dose of 40,000 beads, which became distributed throughout the small airways and parenchyma of the lung. Elastase-coupled beads resulted in a quantifiable loss of alveolar tissue immediately surrounding the beads, an effect that was not observed with beads that lacked protein altogether or with beads containing elastase inactivated by an irreversible inhibitor. Furthermore, beads bound with active elastase elicited local recruitment of mononuclear cells, including macrophages, and polymorphonuclear neutrophils to the site of bead deposition, a feature consistent with the cellular infiltration observed following conventional solubilized elastase challenges. This work identifies a novel bead-based enzyme delivery system that also extends the elastase model of emphysema to permit the characterization of mechanisms that drive alveolar surface area loss following elastin degradation in focal emphysematous lesions.

Inhibition of Notch signalling induces extensive intussusceptive neo‐angiogenesis by recruitment of mononuclear cells

Objectiveto evaluate the effects of Notch inhibition on the vasculature using chick area vasculosa and MxCre Notch1(lox/lox) mice and to test to which extent this effect is mediated by ephrinB2/EphB4.Methods and ResultsInhibition of Notch signaling by Y secretase 1 induced significantly intussusceptive angiogenesis (IA) forming large immature capillary plexus. The latter it is characterized by 40% increased vascular density, pericyte detachment, increased transpermeability and recruitment of mononuclear cells to the sites of new formed transluminal tissue pillars (sign of IA). The process was dramatically enhanced when Notch inhibition was combined with the injection of chicken or mouse bone marrow‐derived mononuclear cells. Along with the morphological alterations of IA, we observed down‐regulation of ephrinB2 mRNA levels. Inhibition of ephrinB2 and EphB4 signaling induces pericytes detachment and significantly upregulation of VEGFR1 but without angiogenic response and recruitment of mononuclear cells. Remarkable, the chemotactic factors SDF‐1 and CXCR4 were up‐regulated only due to the Notch inhibition.ConclusionsOur results indicated that Notch inhibition disturbs vessel stability, induces extravasation of mononuclear cells of bone marrow origin by up‐regulation SDF‐1/CXCR4 chemotactic factors. Recruited cells contributed to formation of transluminal tissue pillars i.e. intussusceptive neo‐angiogenesis, triggering in this way the augmentation of the capillary plexus but without the accompanying vascular maturation and remodeling. .

Implication of lysophosphatidic acid in hypoxic pulmonary vascular remodeling: effects on cell adhesion and migration

Pulmonary hypertension is characterized by cellular and structural changes in vascular wall of pulmonary arteries. Intimal thickening, fibrosis, medial hypertrophy, adventitial fibroblast activation involve progenitors cells and fibrocytes. We hypothesized that lysophosphatidic acid (LPA), a bioactive lipid, is implicated in this vascular remodeling. We investigated the rat model of hypoxic pulmonary hypertension. Chronic hypoxic exposure induced increase of serum levels of circulating LPA (21.6 (11.0 – 42.3) μM vs 40.9 (23.4 – 71.7) μM, p=0.037). In addition, we observed perivascular LPA immunohistochemical staining in lungs of hypoxic rats (antibody against LPA provided by L‐Path Inc.), which co‐localized with lysophospholipase D. Serum from the hypoxic group of animals had significantly higher chemoattractant properties towards rat primary lung fibroblasts, and this increase in cell migration could be prevented by the LPA receptor 1 and 3 antagonist, VPC12249. LPA increased also adhesive properties of human pulmonary artery endothelial cells and of human peripheral blood mononuclear cells via the activation LPA receptor (1 or 3). We suggest that chronic hypoxia increases circulating and tissue level of LPA which might induce fibroblast migration and recruitment of mononuclear cells in pulmonary vasculature that contributes to pulmonary vascular remodeling.

Therapeutic angiogenesis for the critical limb ischemia decreases platelet activation

Platelets are required for the recruitment of bone marrow-derived mononuclear cells (BMMNC) into ischemia-induced vasculature, which underlines their key role in angiogenesis. The difference in platelet immunophenotype between healthy controls and patients with critical limb ischemia (CLI) treated with therapeutic angiogenesis (TA) using BMMNC was assessed. The impact of TA on the expression of platelet membrane markers was studied as well. CLI patients (N = 26) and blood donors as controls (N = 21) were enrolled. Bone marrow (600 ± 50 ml) was centrifuged (3200g, 20 min, 22 °C). BMMNC (100–120 ml) were separated by Optipress I and implanted to the ischemic limb using deep intramuscular injections. Flow cytometry was employed for the peripheral blood platelets immunophenotyping. CD41FITC, CD62PE, CD36FITC, CD29FITC antibodies were used. Patients were followed up prior to the procedure and at months 1, 3 and 6. The expression of CD41 was lower in CLI patients than in the controls. P-selectin (CD62P) was higher in CLI patients than in controls at the baseline and at month 6. It was significantly down-regulated at month 3, however not at months 1 and 6 compared to baseline. Platelet GPIV (CD36) was higher at the baseline, but not during the follow-up compared to the controls. β1-integrin (CD29) progressively decreased during the follow-up as compared to the baseline value. Platelets in CLI express P-selectin, GPIV and β1-integrin more abundantly than platelets of healthy subjects. TA down-regulates the expression of the respective markers. Possible mechanism could be higher clearance of the activated platelets in the ischemic tissues during angiogenesis.

Lymphotoxin Signaling Is Initiated by the Viral Polymerase in HCV-linked Tumorigenesis

Exposure to hepatitis C virus (HCV) typically results in chronic infection that leads to progressive liver disease ranging from mild inflammation to severe fibrosis and cirrhosis as well as primary liver cancer. HCV triggers innate immune signaling within the infected hepatocyte, a first step in mounting of the adaptive response against HCV infection. Persistent inflammation is strongly associated with liver tumorigenesis. The goal of our work was to investigate the initiation of the inflammatory processes triggered by HCV viral proteins in their host cell and their possible link with HCV-related liver cancer. We report a dramatic upregulation of the lymphotoxin signaling pathway and more specifically of lymphotoxin-β in tumors of the FL-N/35 HCV-transgenic mice. Lymphotoxin expression is accompanied by activation of NF-κB, neosynthesis of chemokines and intra-tumoral recruitment of mononuclear cells. Spectacularly, IKKβ inactivation in FL-N/35 mice drastically reduces tumor incidence. Activation of lymphotoxin-β pathway can be reproduced in several cellular models, including the full length replicon and HCV-infected primary human hepatocytes. We have identified NS5B, the HCV RNA dependent RNA polymerase, as the viral protein responsible for this phenotype and shown that pharmacological inhibition of its activity alleviates activation of the pro-inflammatory pathway. These results open new perspectives in understanding the inflammatory mechanisms linked to HCV infection and tumorigenesis.

Interplay between Misplaced Müllerian-Derived Stem Cells and Peritoneal Immune Dysregulation in the Pathogenesis of Endometriosis

In the genetic regulation of Müllerian structures development, a key role is played by Hoxa and Wnt clusters, because they lead the transcription of different genes according to the different phases of the organogenesis, addressing correctly cell-to-cell interactions, allowing, finally, the physiologic morphogenesis. Accumulating evidence is suggesting that dysregulation of Wnt and/or Hox genes may affect cell migration during organogenesis and differentiation of Müllerian structures of the female reproductive tract, with possible dislocation and dissemination of primordial endometrial stem cells in ectopic regions, which have high plasticity to differentiation. We hypothesize that during postpubertal age, under the influence of different stimuli, these misplaced and quiescent ectopic endometrial cells could acquire new phenotype, biological functions, and immunogenicity. So, these kinds of cells may differentiate, specializing in epithelium, glands, and stroma to form a functional ectopic endometrial tissue. This may provoke a breakdown in the peritoneal cavity homeostasis, with the consequent processes of immune alteration, documented by peripheral mononuclear cells recruitment and secretion of inflammatory cytokines in early phases and of angiogenic and fibrogenic cytokines in the late stages of the disease.

Retinoid X Receptor Agonists Impair Arterial Mononuclear Cell Recruitment through Peroxisome Proliferator-Activated Receptor-γ Activation

Mononuclear cell migration into the vascular subendothelium constitutes an early event of the atherogenic process. Because the effect of retinoid X receptor (RXR)α on arterial mononuclear leukocyte recruitment is poorly understood, this study investigated whether RXR agonists can affect this response and the underlying mechanisms involved. Decreased RXRα expression was detected after 4 h stimulation of human umbilical arterial endothelial cells with TNF-α. Interestingly, under physiological flow conditions, TNF-α-induced endothelial adhesion of human mononuclear cells was concentration-dependently inhibited by preincubation of the human umbilical arterial endothelial cells with RXR agonists such as bexarotene or 9-cis-retinoid acid. RXR agonists also prevented TNF-α-induced VCAM-1 and ICAM-1 expression, as well as endothelial growth-related oncogene-α and MCP-1 release. Suppression of RXRα expression with a small interfering RNA abrogated these responses. Furthermore, inhibition of MAPKs and NF-κB pathways were involved in these events. RXR agonist-induced antileukocyte adhesive effects seemed to be mediated via RXRα/peroxisome proliferator-activated receptor (PPAR)γ interaction, since endothelial PPARγ silencing abolished their inhibitory responses. Furthermore, RXR agonists increased RXR/PPARγ interaction, and combinations of suboptimal concentrations of both nuclear receptor ligands inhibited TNF-α-induced mononuclear leukocyte arrest by 60-65%. In vivo, bexarotene dose-dependently inhibited TNF-α-induced leukocyte adhesion to the murine cremasteric arterioles and decreased VCAM-1 and ICAM-1 expression. Therefore, these results reveal that RXR agonists can inhibit the initial inflammatory response that precedes the atherogenic process by targeting different steps of the mononuclear recruitment cascade. Thus, RXR agonists may constitute a new therapeutic tool in the control of the inflammatory process associated with cardiovascular disease.

1088 ESCHERICHIA COLI STRAINS FROM CHRONIC PROSTATITIS PATIENTS INVADE PROSTATE EPITHELIAL CELLS AND INDUCE SECRETION OF INFLAMMATORY MEDIATORS

You have accessJournal of UrologyInfections/Inflammation of the Genitourinary Tract: Prostate & Genitalia1 Apr 20121088 ESCHERICHIA COLI STRAINS FROM CHRONIC PROSTATITIS PATIENTS INVADE PROSTATE EPITHELIAL CELLS AND INDUCE SECRETION OF INFLAMMATORY MEDIATORS Li-Lin Cheng, Laura E. Goeser, Alexandra E. Burleigh, Justin J. Lemke, Rodney A. Welch, and Walter J. Hopkins Li-Lin ChengLi-Lin Cheng Madison, WI More articles by this author , Laura E. GoeserLaura E. Goeser Madison, WI More articles by this author , Alexandra E. BurleighAlexandra E. Burleigh Madison, WI More articles by this author , Justin J. LemkeJustin J. Lemke Madison, WI More articles by this author , Rodney A. WelchRodney A. Welch Madison, WI More articles by this author , and Walter J. HopkinsWalter J. Hopkins Madison, WI More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2012.02.1195AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Category IIIA chronic prostatitis is characterized by the presence of leukocytes in prostatic secretions without culturable bacteria. One possible explanation is that infecting bacteria are harbored within prostate epithelial cells and their intracellular presence induces local secretion of inflammatory mediators. We examined the ability of uropathogenic E. coli to invade mouse prostate epithelial cells in vitro and to induce production of cytokines and chemokines by these cells. METHODS Twenty-four E. coli isolates from chronic prostatitis patients were genotyped for urovirulence factor genes associated with adherence, toxin production, iron acquisition, secretion, and outer membrane proteins. Motility was measured by a swimming assay. Isolates were tested for their invasion of primary epithelial cells derived from the dorsal and lateral prostate lobes of C3H/HeOuJ mice using a gentamicin protection assay. A multiplex assay was used to quantify cytokines/chemokines secreted by cells exposed to E. coli for two hours before addition of gentamicin. RESULTS Among the 24 strains, 10 were intermediate to highly invasive and 14 were low to non-invasive. There was no significant link between any individual gene and invasion intensity; however, a significant positive correlation between motility and epithelial cell invasion was noted. An initial study with eight isolates showed strong responses of IL-6, monocyte chemotactic protein (MCP-1) and granulocyte-macrophage colony stimulating factor (GM-CSF) at 24 hours post exposure, while IL-1β and TNF-α were undetectable. The mediators were induced by seven invasive strains but not by a non-invasive isolate. In addition, strains with cnf-1, cdt, and hlyA toxin genes appeared to induce the least amounts of mediators in prostate epithelial cells. CONCLUSIONS E. coli isolates from chronic prostatitis patients are able to invade prostate epithelial cells with a wide range of intensities. Inflammatory mediators are produced by prostate epithelial cells when exposed to invasive, chronic prostatitis E. coli strains. The secretion of MCP-1, GM-CSF, and IL-6 is associated with recruitment and production of mononuclear inflammatory cells, implying their roles in promoting chronic inflammation. These results thus support a model of chronic prostatic inflammation in the absence of extracellular bacteria, which is the primary characteristic of Category IIIA prostatitis. © 2012 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 187Issue 4SApril 2012Page: e441 Advertisement Copyright & Permissions© 2012 by American Urological Association Education and Research, Inc.MetricsAuthor Information Li-Lin Cheng Madison, WI More articles by this author Laura E. Goeser Madison, WI More articles by this author Alexandra E. Burleigh Madison, WI More articles by this author Justin J. Lemke Madison, WI More articles by this author Rodney A. Welch Madison, WI More articles by this author Walter J. Hopkins Madison, WI More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

HMGB1 promotes recruitment of inflammatory cells to damaged tissues by forming a complex with CXCL12 and signaling via CXCR4

After tissue damage, inflammatory cells infiltrate the tissue and release proinflammatory cytokines. HMGB1 (high mobility group box 1), a nuclear protein released by necrotic and severely stressed cells, promotes cytokine release via its interaction with the TLR4 (Toll-like receptor 4) receptor and cell migration via an unknown mechanism. We show that HMGB1-induced recruitment of inflammatory cells depends on CXCL12. HMGB1 and CXCL12 form a heterocomplex, which we characterized by nuclear magnetic resonance and surface plasmon resonance, that acts exclusively through CXCR4 and not through other HMGB1 receptors. Fluorescence resonance energy transfer data show that the HMGB1-CXCL12 heterocomplex promotes different conformational rearrangements of CXCR4 from that of CXCL12 alone. Mononuclear cell recruitment in vivo into air pouches and injured muscles depends on the heterocomplex and is inhibited by AMD3100 and glycyrrhizin. Thus, inflammatory cell recruitment and activation both depend on HMGB1 via different mechanisms.

Integrated transcriptomic and proteomic evaluation of gentamicin nephrotoxicity in rats

Gentamicin is an aminoglycoside antibiotic, which induces renal tubular necrosis in rats. In the context of the European InnoMed PredTox project, transcriptomic and proteomic studies were performed to provide new insights into the molecular mechanisms of gentamicin-induced nephrotoxicity. Male Wistar rats were treated with 25 and 75 mg/kg/day subcutaneously for 1, 3 and 14 days. Histopathology observations showed mild tubular degeneration/necrosis and regeneration and moderate mononuclear cell infiltrate after long-term treatment. Transcriptomic data indicated a strong treatment-related gene expression modulation in kidney and blood cells at the high dose after 14 days of treatment, with the regulation of 463 and 3241 genes, respectively. Of note, the induction of NF-kappa B pathway via the p38 MAPK cascade in the kidney, together with the activation of T-cell receptor signaling in blood cells were suggestive of inflammatory processes in relation with the recruitment of mononuclear cells in the kidney. Proteomic results showed a regulation of 163 proteins in kidney at the high dose after 14 days of treatment. These protein modulations were suggestive of a mitochondrial dysfunction with impairment of cellular energy production, induction of oxidative stress, an effect on protein biosynthesis and on cellular assembly and organization. Proteomic results also provided clues for potential nephrotoxicity biomarkers such as AGAT and PRBP4 which were strongly modulated in the kidney. Transcriptomic and proteomic data turned out to be complementary and their integration gave a more comprehensive insight into the putative mode of nephrotoxicity of gentamicin which was in accordance with histopathological findings.

Airway Inflammation and IgE Production Induced by Dust Mite Allergen-Specific Memory/Effector Th2 Cell Line Can Be Effectively Attenuated by IL-35

CD4(+) memory/effector T cells play a central role in orchestrating the rapid and robust immune responses upon re-encounter with specific Ags. However, the immunologic mechanism(s) underlying these responses are still not fully understood. To investigate this, we generated an allergen (major house dust mite allergen, Blo t 5)-specific murine Th2 cell line that secreted IL-4, IL-5, IL-10, and IL-13, but not IL-9 or TNF-α, upon activation by the cognate Ag. These cells also exhibited CD44(high)CD62L(-) and CD127(+) (IL-7Rα(+)) phenotypes, which are characteristics of memory/effector T cells. Experiments involving adoptive transfer of this Th2 cell line in mice, followed by three intranasal challenges with Blo t 5, induced a dexamethasone-sensitive eosinophilic airway inflammation. This was accompanied by elevation of Th2 cytokines and CC- and CXC-motif chemokines, as well as recruitment of lymphocytes and polymorphic mononuclear cells into the lungs. Moreover, Blo t 5-specific IgE was detected 4 d after the last intranasal challenge, whereas elevation of Blo t 5-specific IgG1 was found at week two. Finally, pulmonary delivery of the pVAX-IL-35 DNA construct effectively downregulated Blo t 5-specific allergic airway inflammation, and i.m. injection of pVAX-IL-35 led to long-lasting suppression of circulating Blo t 5-specific and total IgE. This model provides a robust research tool to elucidate the immunopathogenic role of memory/effector Th2 cells in allergic airway inflammation. Our results suggested that IL-35 could be a potential therapeutic target for allergic asthma through its attenuating effects on allergen-specific CD4(+) memory/effector Th2 cell-mediated airway inflammation.

Tag SNP Polymorphism of CCL2 and Its Role in Clinical Tuberculosis in Han Chinese Pediatric Population

BackgroundChemokine (C-C motif) ligand 2 CCL2/MCP-1 is among the key signaling molecules of innate immunity; in particular, it is involved in recruitment of mononuclear and other cells in response to infection, including tuberculosis (TB) and is essential for granuloma formation.Methodology/Principal FindingsWe identified a tag SNP for the CCL2/MCP-1 gene (rs4586 C/T). In order to understand whether this SNP may serve to evaluate the contribution of the CCL2 gene to the expression of TB disease, we further analysed distribution of its alleles and genotypes in 301 TB cases versus 338 non-infected controls (all BCG vaccinated) representing a high-risk pediatric population of North China. In the male TB subgroup, the C allele was identified in a higher rate (P = 0.045), and, acting dominantly, was found to be a risk factor for clinical TB (P = 0.029). Homozygous TT genotype was significantly associated with lower CSF mononuclear leukocyte (ML) counts in patients with tuberculous meningitis (TBM) (P = 0.001).Conclusions/SignificanceThe present study found an association of the CCL2 tag SNP rs4586 C allele and pediatric TB disease in males, suggesting that gender may affect the susceptibility to TB even in children. The association of homozygous TT genotype with decreased CSF mononuclear leukocyte (ML) count not only suggests a clinical significance of this SNP, but indicates its potential to assist in the clinical assessment of suspected TBM, where delay is critical and diagnosis is difficult.

Combination of amlodipine and atorvastatin synergistically reduces leukocyte recruitment to mechanically injured mouse femoral artery

Recent studies have demonstrated a potential synergistic effect of the combination of amlodipine with atorvastatin to reduce acute inflammation. The intraluminal wire injury of the mouse femoral artery induced significant leukocyte recruitment to the injured area and oxidative stress within 24 h. Administration of low-dose amlodipine (0.5 mg kg(-1) per day) or atorvastatin (1 mg kg(-1) per day) alone for 7 days failed to modulate leukocyte adhesion, whereas their co-administration for 7 days significantly inhibited leukocyte adhesion. Moreover, flow cytometric analysis showed that injury-induced oxidative stress and CD11b expression in three leukocyte fractions were elevated after injury and then reduced after the co-administration. Next, adoptive transfer of mononuclear cells (MNCs) was performed and MNCs were harvested from mice after wire injury exhibited adhesion to the recipient injured artery. Furthermore, the co-administration of low-dose atorvastatin and amlodipine to MNCs or the vasculature reduced the recruitment of MNCs to the injured artery. Our findings indicate that amlodipine and atorvastatin synergistically inhibit vascular inflammation. The underlying mechanisms of their effect involve, at least in part, stabilizing oxidative stress at the point of injury, suggesting the clinical efficacy of this drug combination for the treatment of vascular diseases.

Targeting the MCP-1/CCR2 System in Diabetic Kidney Disease

Diabetic nephropathy is the leading cause of end-stage renal failure in the Western World and accounts for significant morbidity and mortality in patients with diabetes. Although hyperglycaemia and hypertension are established key determinants in the development of the complication, recent studies suggest that a low-grade inflammatory response may also play a role. Monocyte Chemoattractant Protein 1 (MCP-1), a potent chemokine produced by renal cells, has emerged as a very important player in this process. Specifically, it has been shown that MCP-1 is overexpressed in the kidneys from diabetic animals. Furthermore, there is amelioration of both functional and structural abnormalities in MCP-1- knockout mice in the setting of concomitant diabetes. Over recent years the cellular mechanisms linking MCP-1 to kidney injury have been increasingly delineated and, in particular, it has become evident that MCP-1 contributes to the kidney damage not only by inducing mononuclear cell recruitment, but also by direct activation of resident renal cells. The present review focuses on the most significant advances in understanding the role of MCP-1 in diabetic kidney disease and future potential therapeutic implications.

Potential role for bone marrow-derived fibrocytes in the orbital fibroblast heterogeneity associated with thyroid-associated ophthalmopathy

Fibroblast heterogeneity has been recognized for decades, but the basis for multiple phenotypes among these cells has been investigated only recently. More than 15 years ago, Bucalla and his colleagues described for the first time a population of fibroblast-like cells among circulating mononuclear blood cells. Subsequently these mesenchymal cells, termed fibrocytes, have been characterized and found to participate in normal and pathological tissue remodelling. In this review, I have attempted to present the evidence generated thus far suggesting that fibrocytes are participants in autoimmune diseases where tissues are injured and undergo remodelling. Aspects of their phenotype suggest that they are well suited to help orchestrate immune responses through mononuclear cell recruitment and their ability to produce inflammatory mediators and extracellular matrix molecules. These attributes also raise the possibility that they might be useful targets against which therapeutic agents might be aimed.

Distinct roles for Dectin-1 and TLR4 in the pathogenesis of Aspergillus fumigatus keratitis.

Aspergillus species are a major worldwide cause of corneal ulcers, resulting in visual impairment and blindness in immunocompetent individuals. To enhance our understanding of the pathogenesis of Aspergillus keratitis, we developed a murine model in which red fluorescent protein (RFP)-expressing A. fumigatus (Af293.1RFP) conidia are injected into the corneal stroma, and disease progression and fungal survival are tracked over time. Using Mafia mice in which c-fms expressing macrophages and dendritic cells can be induced to undergo apoptosis, we demonstrated that the presence of resident corneal macrophages is essential for production of IL-1β and CXCL1/KC, and for recruitment of neutrophils and mononuclear cells into the corneal stroma. We found that β-glucan was highly expressed on germinating conidia and hyphae in the cornea stroma, and that both Dectin-1 and phospho-Syk were up-regulated in infected corneas. Additionally, we show that infected Dectin-1−/− corneas have impaired IL-1β and CXCL1/KC production, resulting in diminished cellular infiltration and fungal clearance compared with control mice, especially during infection with clinical isolates expressing high β-glucan. In contrast to Dectin 1−/− mice, cellular infiltration into infected TLR2−/−, TLR4−/−, and MD-2−/− mice corneas was unimpaired, indicating no role for these receptors in cell recruitment; however, fungal killing was significantly reduced in TLR4−/− mice, but not TLR2−/− or MD-2−/− mice. We also found that TRIF−/− and TIRAP−/− mice exhibited no fungal-killing defects, but that MyD88−/− and IL-1R1−/− mice were unable to regulate fungal growth. In conclusion, these data are consistent with a model in which β-glucan on A.fumigatus germinating conidia activates Dectin-1 on corneal macrophages to produce IL-1β, and CXCL1, which together with IL-1R1/MyD88-dependent activation, results in recruitment of neutrophils to the corneal stroma and TLR4-dependent fungal killing.