Cannabinoid signalling in TNF-alpha induced IL-8 release

Original article can be found at: http://www.sciencedirect.com/science/journal/00142999 Copyright Elsevier B.V. DOI : 10.1016/j.ejphar.2006.04.015Peer reviewe

Characterization of the effects of BTK inhibition and monocyte-produced IL-8 on the hematopoietic stem cell niche

Hematopoietic stem and progenitor cells (HSPCs) are multipotent stem cells that give rise to all blood cell lineages. During early zebrafish development, HSPCs interact closely with endothelial cells in an endothelial niche known as the Caudal Hematopoietic Tissue (CHT). How these interactions influence HSPC fate and clonality is not well-understood, however. The chemokine Interleukin-8 (IL-8) may alter HSPC fate by extending the time that HSPCs reside within the CHT. IL-8 is produced by a number of cell types, including monocytes. Using a zebrafish model, we aim to characterize the impact of enforced expression of IL-8 by macrophages on HSPC interactions with the endothelial niche. HSPCs colonize the CHT beginning approximately 32 hours post fertilization before migrating to the kidney marrow at about 6 days post fertilization where they reside for the life of the zebrafish. While in the CHT, HSPCs can be directly imaged by fluorescence microscopy using a fluorescent reporter driven by the HSPC-specific runx1 + 23 enhancer element (runx1+23:GFP). We have generated a transgenic zebrafish line, which ectopically expressesIL-8 in macrophages using the macrophage-specific promoter mpeg1.1 (mpeg1.1:cxcl8 2A mCherry). We hypothesize that the F1 offspring of these lines will show that IL-8 expression by macrophages will increase the residency time of HSPCs in the CHT compared to a non-expressing control group. We are also treating developing fish with Ibrutinib, a therapeutic protein kinase inhibitor that inhibits the IL-8 PI3K/Akt signaling pathway. We hypothesize that HSPCs in treated fish will be fewer in number and will reside in the endothelial niche for less time than non-treated controls. Understanding these interactions will shed light on alterations of HSPC cell fate due to IL-8 expression and provide potential therapeutic targets for patients with hematopoietic disordersNo embargoAcademic Major: Biochemistr

Mycotoxins nivalenol and deoxynivalenol differently modulate cytokine mRNA expression in Jurkat T cells.

Deoxynivalenol (DON) and its hydroxylated form nivalenol (NIV) are Fusarium mycotoxins that occur in cereal grains alone or in combination. Several studies have shown that these metabolites affect lymphocyte functions. However, the molecular mechanisms underlying their activities are still partially known. To address this issue, we examined the influence of NIV and DON in modulating IFNc, IL-2 and IL-8 mRNA levels in Jurkat T cells. In PMA/ionomycin stimulated cells, pre-incubated with increasing concentrations of NIV, transcription was induced in the range 0.06–2 lM; higher concentrations of NIV were found non-stimulating (4 lM) or inhibitory (8 lM) for IFNc and IL-2 whereas IL-8 was still induced. DON administration elicited a similar profile for IL-8 and IFNc, whilst IL-2 mRNA was induced in a broader range of concentrations. Combination of NIV and DON at 1:1 and 1:10 ratios essentially restored the cytokine transcriptional pattern observed with NIV alone but the level of transcripts, with the exception of IL-8, peaked at lower concentrations suggesting interactive effects. Moreover both mycotoxins caused inhibition of cell proliferation, mediated by induction of apoptosis, confirming previous results and highlighting the usefulness of Jurkat as a T-cell model to study the effects of mycotoxins on the immune functions in humans

The phosphodiesterase 5 inhibitor sildenafil decreases the proinflammatory chemokine IL-8 in diabetic cardiomyopathy: in vivo and in vitro evidence

Purpose: Interleukin (IL)-8 is a proinflammatory C-X-C chemokine involved in inflammation underling cardiac diseases, primary or in comorbid condition, such diabetic cardiomyopathy (DCM). The phosphodiesterase type 5 inhibitor sildenafil can ameliorate cardiac conditions by counteracting inflammation. The study aim is to evaluate the effect of sildenafil on serum IL-8 in DCM subjects vs. placebo, and on IL-8 release in human endothelial cells (Hfaec) and peripheral blood mononuclear cells (PBMC) under inflammatory stimuli. Methods: IL-8 was quantified: in sera of (30) DCM subjects before (baseline) and after sildenafil (100 mg/day, 3-months) vs. (16) placebo and (15) healthy subjects, by multiplatform array; in supernatants from inflammation-challenged cells after sildenafil (1 µM), by ELISA. Results: Baseline IL-8 was higher in DCM vs. healthy subjects (149.14 ± 46.89 vs. 16.17 ± 5.38 pg/ml, p < 0.01). Sildenafil, not placebo, significantly reduced serum IL-8 (23.7 ± 5.9 pg/ml, p < 0.05 vs. baseline). Receiver operating characteristic (ROC) curve for IL-8 was 0.945 (95% confidence interval of 0.772 to 1.0, p < 0.01), showing good capacity of discriminating the response in terms of drug-induced IL-8 decrease (sensitivity of 0.93, specificity of 0.90). Sildenafil significantly decreased IL-8 protein release by inflammation-induced Hfaec and PBMC and downregulated IL-8 mRNA in PBMC, without affecting cell number or PDE5 expression. Conclusion: Sildenafil might be suggested as potential novel pharmacological tool to control DCM progression through IL-8 targeting at systemic and cellular level

Flagellin induces β-defensin 2 in human colonic ex vivo infection with enterohemorrhagic Escherichia coli

Enterohemorrhagic E. coli (EHEC) is an important foodborne pathogen in the developed world and can cause life-threatening disease particularly in children. EHEC persists in the human gut by adhering intimately to colonic epithelium and forming characteristic attaching/effacing lesions. In this study, we investigated the innate immune response to EHEC infection with particular focus on antimicrobial peptide and protein expression by colonic epithelium. Using a novel human colonic biopsy model and polarized T84 colon carcinoma cells, we found that EHEC infection induced expression of human β-defensin 2 (hBD2), whereas hBD1, hBD3, LL-37 and lysozyme remained unchanged. Infection with specific EHEC deletion mutants demonstrated that this was dependent on flagellin, and apical exposure to purified flagellin was sufficient to stimulate hBD2 and also interleukin (IL)-8 expression ex vivo and in vitro. Flagellin-mediated hBD2 induction was significantly reduced by inhibitors of NF-κB, MAP kinase p38 and JNK but not ERK1/2. Interestingly, IL-8 secretion by polarized T84 cells was vectorial depending on the side of stimulation, and apical exposure to EHEC or flagellin resulted in apical IL-8 release. Our results demonstrate that EHEC only induces a modest immune response in human colonic epithelium characterized by flagellin-dependent induction of hBD2 and low levels of IL-8

Зголемени вредности на IL-8 во ран втор триместар и нивната поврзаност со предвремено раѓање

Cytokines (IL-1, IL-6, IL-8, TNF- alfa) are of crucial importance during pregnancy; they are produced by the placenta in the amniotic fluid and they are elevated in case of intrauterine inflammation. The аim of the study was to prove the ratio between the increased IL-8 in the amniotic fluid in the beginning of the second trimester (16-22 g.w.) and premature birth (< 36.6 g.w.). Material and methods: This was a prospective study that included 150 pregnant patients that had clinical indication for amniocentesis (advanced mother’s age, abnormal test of PRISCA I, suspicious anomalies of the fetus, virus infection or mother’s wish). They all gave a signed consent on being informed about the aims of the study, and following the protocol, they were analyzed and examined i.e. all patients underwent ultrasound examination, vaginal cervicometry. Five ml. of amniotic fluid during the process of amniocentesis was taken for the purpose of the study. All patients were followed until they gave birth, and the exact week of gestation was noted and compered with the IL-8 level. Results: All 150 patients were in the period of 16th-22nd gestational weeks. Twenty of the total of 150 patients had preterm delivery. A total of 139 patients conceived naturally and 9 patients underwent in vitro fertilisation (IVF) and embryo transfer (ET). In those with IVF and ET, 3 had preterm birth. 80% of patients that had preterm birth had increased IL- 8 levels. Median cervical length in those who gave birth at term was 32.1 mm and in those who gave preterm birth was 30.7mm. Conclusion: The study has confirmed the reason for examining cytokines as a method of discovering asymptomatic changes in patients who would give a premature birth.Цитокините (IL-1, IL-6, IL-8, TNF- Alfa) се од исклучително значење во бременоста и тие се продуцираат од страна на постелката во амнионската течност и се зголемени доколку постои интраутерина инфламација. Целта на студијата беше да се докаже соодносот на покаченото ниво на IL-8 во амнионската течност во почетокот на раниот втор триместар (16-22 г.н.) и предвременото породување (< 37 г.н.). Материјал и методи: Во оваа проспективна студија беа вклучени 150 гравидни пациентки, каде постоеше медицинска индикација за изведување на амниоцентеза (напредната мајчина возраст, висок ризик на PRISCA I, суспектни аномалии на фетусот, вирусни инфекции, или по желба на мајката). По потпишана согласност за учество во студијата, сите пациентки беа анализирани, односно на сите пациентки им беше направен ехо преглед, вагинална цервикометрија, и беа земени дополнителни 5 мл амнионска течност при изведување на амниоцентезата. Сите пациентки беа следени сè до нивното породување, каде точно беше нотирана гестациската недела на породување, а потоа споредена со нивото на IL-8. Резултати: Сите 150 пациентки беа во периодот од 16-22 гестациска недела. Кај 20 од вкупно 150 пациентки констатиравме предвремено раѓање, додека, пак, 120 пациентки се породија во термин. 139 пациентки имаа зачнато природно, додека 9  со ИВФ и ЕТ, од кои три се породија предвремено. 80% од пациентките кои се породија предвремено имаа зголемени вредности на IL-8. Средната вредност на должина на цервиксот кај оние кои се породија во термин беше 32,1 мм, додека кај оние превремено породени беше 30,7 мм. Заклучок: Оваа студија јапотврди оправданоста за испитување цитокини како метод за откривање на асимптоматски промени кај пацинтки кое ќе се породат предвремено. &nbsp

Using Interleukin 6 and 8 in Blood and Bronchoalveolar Lavage Fluid to Predict Survival in Hematological Malignancy Patients With Suspected Pulmonary Mold Infection.

Background: Molds and other pathogens induce elevated levels of several cytokines, including interleukin (IL)-6 and IL-8. The objective of this study was to investigate the prognostic value of IL-6 and IL-8 as well as fungal biomarkers in blood and bronchoalveolar lavage fluid (BAL) for overall survival in patients with underlying hematological malignancies and suspected mold infection. Methods: This cohort study included 106 prospectively enrolled adult cases undergoing bronchoscopy. Blood samples were collected within 24 h of BAL sampling and, in a subset of 62 patients, serial blood samples were collected up until 4 days after bronchoscopy. IL-6, IL-8, and other cytokines as well as galactomannan (GM) and β-D-glucan (BDG) were assayed in blood and BAL fluid and associations with overall mortality were assessed at the end of the study using receiver operating characteristic (ROC) curve analysis. Results: Both blood IL-8 (AUC 0.731) and blood IL-6 (AUC 0.699) as well as BAL IL-6 (AUC 0.763) and BAL IL-8 (AUC 0.700) levels at the time of bronchoscopy were predictors of 30-day all-cause mortality. Increasing blood IL-6 levels between bronchoscopy and day four after bronchoscopy were significantly associated with higher 90-day mortality, with similar findings for increasing IL-8 levels. In ROC analysis the difference of blood IL-8 levels between 4 days after bronchoscopy and the day of bronchoscopy had an AUC of 0.829 (95%CI 0.71-0.95; p < 0.001) for predicting 90-day mortality. Conclusions: Elevated levels of IL-6 and IL-8 in blood or BAL fluid at the time of bronchoscopy, and rising levels in blood 4 days following bronchoscopy were predictive of mortality in these patients with underlying hematological malignancy who underwent bronchoscopy for suspected mold infection

Treponema pallidum (syphilis) antigen TpF1 induces angiogenesis through the activation of the IL-8 pathway

open15Over 10 million people every year become infected by Treponema pallidum and develop syphilis, a disease with broad symptomatology that, due to the difficulty to eradicate the pathogen from the highly vascularized secondary sites of infection, is still treated with injections of penicillin. Unlike most other bacterial pathogens, T. pallidum infection produces indeed a strong angiogenic response whose mechanism of activation, however, remains unknown. Here, we report that one of the major antigen of T. pallidum, the TpF1 protein, has growth factor-like activity on primary cultures of human endothelial cells and activates specific T cells able to promote tissue factor production. The growth factor-like activity is mediated by the secretion of IL-8 but not of VEGF, two known angiogenic factors. The pathogen’s factor signals IL-8 secretion through the activation of the CREB/NF-κB signalling pathway. These findings are recapitulated in an animal model, zebrafish, where we observed that TpF1 injection stimulates angiogenesis and IL-8, but not VEGF, secretion. This study suggests that the angiogenic response observed during secondary syphilis is triggered by TpF1 and that pharmacological therapies directed to inhibit IL-8 response in patients should be explored to treat this disease.openPozzobon, Tommaso; Facchinello, Nicola; Bossi, Fleur; Capitani, Nagaja; Benagiano, Marisa; DI BENEDETTO, Giulietta; Zennaro, Cristina; West, Nicole; Codolo, Gaia; Bernardini, Marialina; Baldari, Cosima Tatiana; D'Elios, Mario Milco; Pellegrini, Luca; Argenton, Francesco; DE BERNARD, MarinaPozzobon, Tommaso; Facchinello, Nicola; Bossi, Fleur; Capitani, Nagaja; Benagiano, Marisa; DI BENEDETTO, Giulietta; Zennaro, Cristina; West, Nicole; Codolo, Gaia; Bernardini, Marialina; Baldari, Cosima Tatiana; D'Elios, Mario Milco; Pellegrini, Luca; Argenton, Francesco; DE BERNARD, Marin

PENGARUH SUPLEMENTASI ISOFLAVON KEDELAI TERHADAP WANITA PENDERITA AKNE VULGARIS : Kajian Jumlah Lesi, Dihydrotestosterone, Toll-Like Receptor-2, dan Interleukin-8

Latar belakang : Akne vulgaris (AV) merupakan penyakit inflamasi kulit yang paling sering dijumpai. Isoflavon kedelai telah terbukti sebagai antiandrogen dan antiinflamasi. Tujuan penelitian ini membuktikan pengaruh isoflavon kedelai terhadap lesi AV akibat penurunan kadar dihydrotestosterone (DHT), Toll-like receptor-2 (TLR-2), Interleukin-8 (IL-8) pada wanita penderita AV. Metoda : Randomized pre and post test control design. Penelitian pendahuluan dengan besar sampel 25 orang, dirandomisasi dalam kelompok plasebo, isoflavon 40 mg, 80 mg, 120 mg, 160 mg, lama penelitian 4 minggu. Penelitian lanjutan dengan 40 sampel, dirandomisasi dalam kelompok kontrol dan perlakuan, lama penelitian 12 minggu. Variabel bebas isoflavon kedelai, varibel terikat lesi AV dan variabel antara adalah DHT, TLR-2, dan IL-8. Hasil : Lesi AV pada awal penelitian pendahuluan 100,178,92 dan akhir penelitian 78,945,85 terdapat penurunan bermakna (t:2,525)(p:0,019). Delta kelompok isoflavon 160 mg lebih besar dari plasebo, isoflavon 40 mg, 80 mg, dan 120 mg (Z:-2,611)(p: 0,009). Lesi AV pada awal penelitian lanjutan 97,8547,614 dan akhir penelitian 59,3845,549, terdapat penurunan yang bermakna (Z:-4,300)(p:0,000). DHT awal penelitian 307,6150,38 pg/ml dan akhir penelitian 283,5253,13 pg/ml, terdapat penurunan yang bermakna (Z:-2,204)(p:0,027). TLR-2 awal penelitian 4539,82862 pg/ml dan akhir penelitian 3944,63592,42 pg/ml terdapat penurunan bermakna (Z:-3,624)(p:0,000). IL-8 awal penelitian 411,3167,137 pg/ml dan akhir penelitian 311,09103,917 pg/ml, terdapat penurunan bermakna (Z:-4,557) (p:0,000). Simpulan : Pemberian suplementasi isoflavon kedelai dengan variasi dosis selama 4 minggu, dapat menurunkan lesi AV dan diperoleh dosis yang paling baik yaitu dosis 160 mg serta pemberian selama 12 minggu menyebabkan penurunan bermakna terhadap lesi AV, DHT, TLR-2, dan IL-8 dibandingkan terapi standar. Kata kunci : Akne vulgaris, isoflavon kedelai, lesi AV, DHT, TLR-2, IL-8. Background. Acne vulgaris (AV) is the most common feature of skin inflammatory diseases. Soy isoflavone has proven as an anti-androgen and an anti-inflammation. The purpose of this research was to prove the effect of soy isoflavone in AV lesion that caused by the decreased level of dihydrotestoreone (DHT), Toll-like receptor-2 (TLR-2), Interleukin-8 (IL-8) in women with AV. Methods. This research used randomized pre and post test control design. The first research had 25 samples, randomized in 5 groups: placebo, isoflavone 40 mg, 80 mg, 120 mg, 160 mg group in 4 weeks length of research. The last research had 40 samples, randomized in controled and threated group in 12 weeks length of research. The independent variables was soy isoflavone, the dependent variables was AV, the confounding variables were DHT, TLR-2, and IL-8. Results. There was a significant degradation (t=2.525; p=0.019) of AV lesion in the early (100.1+78.92) and in the end of the first research (78.9+45.85). Delta of isoflavone 160 mg group more than other groups (Z=-2.61; p=0.009). There was a significant degradation (Z=-4.300; p=0.000) of AV lesion in the early (97.85+47.614) and in the end of the last research (59.38+45.5). DHT level had a signficant decrease (Z=-2.204; p=0.027) from the first research (307.6+150.38 pg/ml) to the last research (283.5+253.13 pg/ml). TLR-2 level had a signficant decrease (Z=-3.624; p=0.000) from the first research (4539.8+2862 pg/ml) to the last research (3944.6+3592.42 pg/ml). IL-8 level had a signficant decrease (Z=-4.557; p=0.000) from the first research (411.31+67.137 pg/ml) to the last research (311.09+103.917 pg/ml). Conclusion. The administration of soy isoflavone with variances of doses in 4 weeks can decrease AV lesion and the best dose was 160 mg, and the administration in 12 weeks can make significant degradation of AV lesion and significant decreased level of DHT, TLR-2, and IL-8 compared by standard therapy. Keywords: Acne vulgaris, soy isoflavone, AV lesion, DHT, TLR-2, IL-