bluuebunny/crossref_metadata_2025_split huggingface dataset:
P012 α4β7 Receptor occupancy by vedolizumab at trough time-point occurs at the membranal level and is not associated with endoscopic healing in inflammatory bowel disease patients
Authors: Ungar B , Ben-Shatach Z , Haj-Natour O , Yavzori M , Fudim E , Picard O , Coscas D , Lahat A , Lidar M , Eliakim R , Kopylov U , Ben-Horin S 
Published: January 2020

Abstract

Background
Previous studies have shown inconclusive associations between vedolizumab trough levels and therapy outcomes. A near-complete occupancy of blood and intestinal α4β7 was exerted by vedolizumab through a wide-range of drug levels, regardless of clinical outcomes. However, these observations were not conducted specifically at the time of trough drug levels, did not include α4β7 highly expressing eosinophils and did not establish whether occupancy is in fact due to fully internalised VDZ- α4β7 complex in the target T-cell.

Methods
Peripheral blood (PB) and intestinal lamina propria (LP) were collected from IBD patients undergoing lower endoscopies on the day of vedolizumab infusion (trough time-point). Clinical and endoscopic scores were prospectively recorded. FACS analysis was performed using fluorescent conjugated-vedolizumab to investigate α4β7 occupancy on effector-memory T cells (CD3+CD45RO+ cells) and on eosinophils (CD49d+CD294+) in PB. Using a newly-developed dissociation assay, we also assessed the degree of membrane-bound VDZ-mediated blocked- α4β7.

Results
Forty-five IBD patients (16, 35% Crohn’s patients, 18, 40% in endoscopic remission) underwent clinical and endoscopic assessment just before their scheduled vedolizumab infusions. The median percentage of free α4β7 receptors at trough was 1.3% (IQR 0.5–3.7%) for PB T cells and 7.2% (IQR 3.6–23.4%) for LP T cells. No statistically significant difference was detected in α4β7 receptor saturation between patients with and without mucosal healing, both for PB (free α4β7 receptors: median 0.5%, 1.46%, IQR 0.3–1.4%, 0.9–3.5% respectively, p = 0.15) and for LP (free α4β7 receptors, median: 11.5%, 7%, IQR 3.5–24.2%, 5.8–20.9%, p = 0.88). Similarly, almost complete α4β7 receptor saturation at trough was detected among PB eosinophils (1.5% free α4β7 receptors, IQR 0.9–2.6%). Upon dissociation assay, a significant number of membrane-bound α4β7 receptors became detectable both on PB and LP T cells, with medians of 31% (IQR 24–41%), 22.2% (IQR 15–31%) respectively, and also for eosinophils, with a median of 83% (IQR 67–89%). When compared with values before dissociation, the proportion of unbound receptors was considerably lower (p &lt; 0.001 for all analyses, respectively).

Conclusion
Even at trough, almost complete α4β7 receptor saturation is reached, both for PB and LP T cells, as well as for PB eosinophils, regardless of endoscopic activity status. This receptor occupancy is unrelated to receptor-internalisation and reflects a complete and continuous antigen-binding at the membranal level.
P017 C86/CD16 macrophages accumulate in the mucosa of B3 patients and could mediate EMT in Crohn’s disease
Authors: Boronat-Muñoz A , Cejudo-Garces A , Lledo-Gil P , Cosín-Roger J , Coll S , Navarro-Vicente F , Alos R , Calatayud S , Barrachina M D , Ortiz-Masiá D 
Published: January 2020

Abstract

Background
Macrophages contribute to fibrosis through the release of different mediators and the pattern of secretion may vary according to their phenotype.

Methods
The aim of the present study is to analyse the pattern of expression of macrophages, of EMT-related genes and cytokines in surgical resections from Crohn’s disease (CD, n = 43) patients which were categorised according to Montreal classification (B2 or B3); unaffected mucosa of patients with ileocecal cancer was used as control (n = 20). mRNA was isolated from intestinal samples and the expression of macrophage, EMT markers and cytokines were analysed by RT-PCR. PBMCS were isolated from healthy donors and treated during 5 days with secretomes, from control, B2 or B3 surgical resections; the mRNA expression of macrophage markers was determined by RT-PCR. U937 cells were differentiated to macrophages and then treated with IFNγ (20 ng/ml) for 4 days, the mRNA expression of macrophages markers were determined by RT-PCR. Results are expressed as mean ± SEM (n ≥ 5). Statistical analysis was performed by ANOVA + Newman–Keuls or t-test. Correlations between data were analysed using Pearson’s correlation coefficient (*p &lt; 0.05).

Results
The expression of CD16 and CD86 was significantly higher in intestinal samples from B3 CD patients (7.2 ± 1.1 and 7.7 ± 1.3, respectively) than in controls (1.4 ± 0.2 and 2.5 ± 0.4, respectively) or B2 CD patients (4.8 ± 0.9 and 4.5 ± 0.6, respectively). The mRNA expression of CD16 and CD86 were significantly higher in PBMCS treated with B3-secretomes than in those treated with B2- or control secretomes. The expression of CD16 and CD86 significantly correlated with FSP1 (r = 0.74, p = 0.002* and r = 0.66, p = 0.003*, respectively), VIMENTIN (r = 0.60, p = 0.02* and r = 0.82, p = 0.001*, respectively), SNAIL1 (r = 0.61, p &lt; 0.01* r = 0.52, p = 0.04*, respectively), IL4 (r = 0.63, p = 0.01* and r = 0.60, p = 0.02*, respectively) and IFNγ (r = 0.56, p = 0.001* and r = 0.58, p = 0.01*, respectively) in intestinal tissue from the fistulising CD group. U937 cells treated with IFNγ increased significantly the mRNA expression of CD16 (1.94 ± 0.24* vs. vehicle) and CD86 (1.60 ± 0.17* vs. vehicle).

Conclusion
A macrophage phenotype expressing CD86/CD16 may act as a source of EMT mediators in intestinal tissue from CD patients with a penetrating (B3) behaviour. IFNγ could be responsible for the increase in the number of CD86/CD16 macrophages in the B3 behaviour.
P018 Per- and polyfluoroalkyl substances (PFAS) are significantly increased in patients with late-onset of ulcerative colitis
Authors: Fart F , Salihovic S , McGlinchey A , Orešič M , Halfvarson J , Hyötyläinen T , Schoultz I 
Published: January 2020

Abstract

Background
Environmental factors have been implicated in the pathogenesis of inflammatory bowel disease (IBD), particularly late onset disease. Per- and polyfluoroalkyl substances (PFAS) are man-made chemicals with a long biological half-life that have been extensively used since the 1950s and have been proposed to interfere with the bile acid synthesis. Therefore, to investigate if late onset IBD correlates with higher PFAS levels, we measured serum levels of PFAS and bile acids in patients diagnosed with IBD later in life.

Methods
Serum samples were collected from patients diagnosed with ulcerative colitis (n = 20) and Crohn’s disease (n = 20) at the age of ≥55 years. Blood donors (n = 20) were used as healthy controls and were matched by gender and age. The levels of PFAS and bile acids were assessed by ultra performance liquid chromatography coupled to a triple quadrupole mass spectrometer.

Results
The total amount of PFAS was significantly higher in patients with ulcerative colitis compared with healthy controls (p = 0.021) or patients with Crohn’s disease (p = 0.015). No difference was found in total PFAS levels between Crohn’s disease patients and healthy controls (p = 0.841). Seven out of 30 bile acids correlated to the total PFAS level.

Conclusion
Our results demonstrate that PFAS levels are increased in patients with late-onset of ulcerative colitis compared with Crohn’s disease patients and healthy controls. This finding indicates that PFAS might represent an environmental risk factor for ulcerative colitis. However, additional studies assessing the functional consequences of increased PFAS in late-onset ulcerative colitis are required to confirm this hypothesis.
P024 Evaluation of the clinical course of patients diagnosed with primary sclerosing cholangitis: single centre study
Authors: Wiecek S , Wojtyniak A , Pindur B , Grzybowska-Chlebowczyk U 
Published: January 2020

Abstract

Background
Primary sclerosing cholangitis (PSC) is a rare liver cholestatic disease with unknown aetiology, severe clinical course and poor prognosis. PSC most often co-occurs with inflammatory bowel diseases (IBD), especially ulcerative colitis (UC). The aim of the study. We aimed to analyse the clinical course of patients diagnosed with PSC hospitalised in the Department of Gastroenterology, Medical University of Silesia in Katowice in 2009–2018.

Methods
The analysis included 30 patients, aged from 7 to 18 years (mean age 13.5 years), 21/30 boys (70%) and 9/30 girls (30%), diagnosed with PSC. The analysis included age of diagnosis, clinical symptoms, course of the disease, laboratory and imaging results and complications.

Results
The average diagnosis age was 13 years. 22/30(73.3%) patients suffered from UC and 4/30 (13.3%) were diagnosed with Crohn’s disease. 2/30 (6.66%) patients remained IBD-free. Among them 7/30(23.3%) had an overlap syndrome—primary sclerosing cholangitis and autoimmune hepatitis. At the time diagnosis 6/30 (20%) patients presented abdominal pain, which was the most common symptom, while 16/30 (57%) were asymptomatic but had abnormal laboratory test results. When PSC was detected before IBD, patients had complications more often compared with those diagnosed with IBD first or PSC and IBD at the same time. Moreover, complications were observed more often in patients with changes characteristic of PSC visible in their first USG than in patients without mentioned abnormalities in first USG.

Conclusion
Patients diagnosed with PSC before IBD diagnosis are more likely to have a more aggressive course of the disease. Monitoring liver markers in IBD patients is important since most PSC cases are asymptomatic and their elevation might be the first sign of the disease.
P032 Increased endoplasmic reticulum stress-specific chaperones characterise CD fibrosis epithelium tissues and participate to in vitro induction of intestinal fibrobast differentiation
Authors: Vieujean S , Hu S , Bequet E , Salee C , Massot C , Bletard N , Pierre N , Quesada calvo F , Baiwir D , Mazzucchelli G , De Pauw E , Delvenne P , Meuwis M A , Louis E 
Published: January 2020

Abstract

Background
Intestinal fibrosis is a complication of Crohn’s disease (CD) characterised by myofibroblasts and extracellular matrix accumulation within the submucosa and smooth muscles, leading to bowel strictures. No medical treatment exists to treat or reverse intestinal fibrosis leading often to surgical resection. The potential role of intestinal epithelium in the fibrotic process remains poorly defined.

Methods
We performed a pilot study on ileal fibrostricturing CD surgical samples (n = 5), comparing the proteome of surface epithelium isolated by laser capture microdissection in normal and fibrotic zones. Confirmation of the specific protein increases was obtained by immunohistochemistry in colonic and ileal samples of CD (n = 44) compared with healthy subjects (n = 40), as well as in intestinal epithelial cell line under induced endoplasmic reticulum (ER) stress. A model of fibroblast to myofibroblast differentiation induction was also challenged using preconditioned media of intestinal epithelial cells after a pulsed ER stress.

Results
Label-free proteomics revealed high ER stress in the epithelium surrounding fibrotic bowel wall, involving anterior gradient protein 2 homolog (AGR2) and 78 kDA glucose-regulated protein (BiP). Confirmation of both proteins increase was obtained by immunohistochemistry. ER stress induction in intestinal epithelial cells was associated with an intracellular increase of AGR2, BiP and ER stress markers as sXPB1 and CHOP. AGR2 was also detected in the culture medium of these epithelial cells and myofibroblast differentiation was obtained using this culture medium.

Conclusion
The increase of ER stress proteins observed in fibrostenosing tissues together with these preliminary evidences of fibroblast to myofibrobast differentiation obtained by paracrine action of intestinal epithelial cell preconditioned to ER stress induction suggest a role of epithelial ER stress in Crohn’s disease intestinal fibrosis.
P033 Combination of vedolizumab with tacrolimus is more efficient in reducing intestinal inflammation than vedolizumab alone in the DSS-induced acute colitis mouse model
Authors: Manzini R , Atrott K , Schwarzfischer M , Laimbacher A , Lang S , Rogler G , Scharl M , Spalinger M R 
Published: January 2020

Abstract

Background
The humanised monoclonal antibody vedolizumab is used in the treatment of ulcerative colitis (UC) and Crohn’s disease (CD). Vedolizumab reduces intestinal inflammation through inhibition of the integrin heterodimer α 4β 7, responsible for the homing of T cells to the intestinal mucosa. Recent studies have also shown a possible involvement of vedolizumab in the regulation of the innate immune system. Particularly in CD, only a fraction of patients respond to vedolizumab treatment, and combination therapy with immunosuppressant drugs, such as the calcineurin-inhibitor tacrolimus, might prove beneficial. The aim of this study was to assess if co-treatment of vedolizumab and tacrolimus is more efficient in reducing intestinal inflammation in an acute colitis mouse model and to unravel the underlying molecular mechanisms.

Methods
NOD-SCID-SGM3 mice were reconstituted with human CD34+ cells and treated with 3% dextrane sodium sulphate (DSS) in drinking water to induce acute colitis. Mice were treated with vedolizumab alone (30mg/kg, inject 3 days prior to DSS-start and 50mg/kg at day 0 and at day 4 of DSS-treatment), tacrolimus alone (1mg/kg/day intraperitoneally), or a combination of tacrolimus and vedolizumab during colitis induction.

Results
As expected, DSS-treatment induced colitis in mice as observed by weight loss, diarrhoea, colon shortening, and endoscopic signs of inflammation categorised by the MEICS score. This translated histologically to an increased immune cell infiltration and epithelial erosion. Vedolizumab and tacrolimus treatment alone did not significantly reduce colitis severity, although endoscopy showed slightly less severe inflammation in vedolizumab-treated mice. Combination of vedolizumab and tacrolimus, however, clearly reduced colonoscopy and histology scores. DSS-treatment increased the number of CD3 T cells and CD68 macrophages in the intestine, an effect counteracted by vedolizumab or tacrolimus alone and further pronounced by combination treatment. Particularly vedolizumab treatment, either alone or in combination, caused clear reduction of pro-inflammatory M1 macrophages. Additionally, vedolizumab alone or combined decreased the levels of intestinal epithelial cell apoptosis as indicated by staining for cleaved caspase-3.

Conclusion
Our data demonstrate that the anti-inflammatory effect of vedolizumab is potentiated by co-treatment with tacrolimus. Notably, the combination of both drugs was more efficient in reducing T-cell and macrophage infiltration into the intestine. This indicates that the combination of vedolizumab with immunosuppressant drugs might prove beneficial for patients that do not respond to vedolizumab-only therapy.
P037 Antagonism of chemokine receptor CCR9 synergises with anti-TNFα immunotherapy to reduce inflammation in the MDR1a−/− mouse model of colitis
Authors: Campbell J , Ertl L , Ebsworth K , McMahon J , Zhang P , Chhina V , Charo I , Schall T 
Published: January 2020

Abstract

Background
The development of anti-TNFα immunotherapies has led to great advances in quality of life for Crohn’s and colitis patients and has allowed a reduction in steroid use for these diseases. However, a significant proportion of IBD patients is resistant to anti-TNFα agents, or becomes resistant to such treatments over time. Preclinical research and clinical trials have suggested that CCR9 antagonism can effectively reduce colon inflammation. CCR9 is expressed on a subset of T cells dedicated to monitoring the immune status of the intestine. When activated by its only known chemokine ligand, CCL25/TECK, CCR9 triggers the adhesion of gut-specific T cells to vascular endothelium and the subsequent ingress of these cells to intestinal tissues. Antagonism of this receptor inhibits the accumulation of T cells within inflamed intestinal epithelium and lamina propria. Since anti-TNFα and CCR9 antagonists act through entirely different mechanisms, we asked whether these two approaches might synergise and expand the effectiveness and scope of each therapy.

Methods
We used a piroxicam-accelerated version of the MDR1a−/− spontaneous colitis model in mice. In this model, 7week MDR1a−/− mice (or WT FVB controls) receive piroxicam in powdered chow for 10 days. Mice are monitored both during and after the piroxicam feeding period, for a total of 21 days. Treatment groups (eight mice each) include an antibody (a saturating dose of either anti-TNFα or its isotype-matched control) and a CCR9 antagonist (or its vehicle alone as a control). Body weight and diarrhoea score were recorded throughout the entire 21 days period, after which colons were obtained from sacrificed animals for weighing, length measurement and imaging.

Results
We have found that combined dosing of piroxicam-fed MDR1a−/− mice with a CCR9 antagonist and an anti-TNFα MAb significantly reverses loss of body weight, diarrhoea score and colon inflammation. The combined dosing was more effective than anti-TNFα or CCR9 antagonist alone, as measured by colon length/weight ratio, neutrophil accumulation in the colonic epithelium and reversal of the loss in body weight characteristic of this model.

Conclusion
These data suggest that combining a CCR9 antagonist with existing anti-TNFα treatment regimens may extend the benefits of anti-TNFα immunotherapy to a larger class of patients suffering from IBD and may prolong the effectiveness of anti-TNFα immunotherapy for those patients already undergoing such treatments.
P041 Melatonin controls microbiota in colitis through Toll-like receptor 4 signalling
Authors: Park Y S , KIm S W , Cheon J H 
Published: January 2020

Abstract

Background
Microbial dysbiosis has long been postulated to be associated with the pathogenesis of inflammatory bowel disease (IBD). Although evidence supporting the anti-colitic effects of melatonin have been accumulating, it is not clear how melatonin affects the microbiota. Herein, we investigated the effects of melatonin on the microbiome in colitis and identified involvement of Toll-like receptor (TLR) 4 signalling in the effects.

Methods
Colitis was induced in 8-week-old wild-type (WT) and TLR4 KO mice by administration of 2.5% dextran sulphate sodium (DSS) in drinking water and intraperitoneal (i.p.) treatment with melatonin (Mel) or vehicle (Veh). Body weight changes were monitored daily.

Results
DSS-treatment for 10 days significantly increased body weight loss in the vehicle-treated group of both wild-type and TLR4 KO mice. Melatonin improved dextran sulphate sodium (DSS)-induced colitis and reverted microbial dysbiosis in wild-type (WT) mice but not in TLR4 knockout (KO) mice. Induction of goblet cells was observed with melatonin administration, which was accompanied by suppression of Il1b and Il17a and induction of melatonin receptor and Reg3β, an antimicrobial peptide (AMP) against Gram-negative bacteria. In vitro, melatonin treatment of HT-29 intestinal epithelial cells promotes mucin and wound healing and inhibits growth of Escherichia coli.

Conclusion
we showed that melatonin significantly increases goblet cells, Reg3β and the ratio of Firmicutes to Bacteriodetes by suppressing Gram-negative bacteria through TLR4 signalling. Our study suggests that sensing of bacteria through TLR4 and regulation of bacteria through altered goblet cells and AMPs is involved in the anti-colitic effects of melatonin. Melatonin may have use in therapeutics for IBD.
P047 Differences in extra cellular matrix remodelling in highly active Crohn’s disease and ulcerative colitis
Authors: Domislović V , Mortensen J H , Karsdal M A , Brinar M , Manon-Jensen T , Krznarić Ž 
Published: January 2020

Abstract

Background
Ulcerative colitis (UC) and Crohn’s disease (CD) require continuous evaluation of disease activity and response to therapy. Current gold standard is endoscopy; therefore it is important to investigate biomarkers associated with endoscopic disease activity. Extra cellular matrix (ECM) consists of basement membrane (BM) and interstitial matrix (IM). BM consists mainly of type IV collagen, while IM of types I, III and V collagen. Pathological environment leads to impaired remodelling, structure, quality and function of the ECM. We investigated biomarkers of collagen degradation and formation and their association with endoscopic disease activity and in patients with CD and UC.

Methods
In this cross-sectional study, we measured six biomarkers of ECM remodelling in 94 IBD patients (60 with CD and 34 UC). Biomarkers of type III collagen degradation (C3M) and formation (PRO-C3), type IV collagen degradation (C4M), formation (PRO-C4), type V collagen formation (PRO-C5) and PROC23 were measured in serum by ELISA. Inflammatory activity was measured using endoscopic scores; SES-CD for CD, MES and modified MES (mMES) for UC. Patients were divided in remission and mild activity group vs. moderate-to-severe activity (SES-CD &lt; 3 vs. ≥3 and MES &lt; 2 vs. ≥2) Student t-test and correlation analysis were applied in statistical analysis.

Results
CD patients with moderate and severe disease had lower levels of PRO-C3 (14.7 ± 10.5 vs. 9.9 ± 4.6, p = 0.04), and higher levels of C4M (26.54 ± 10.5 vs. 37.8 ± 20, p = 0.009) compared with patients in remission and mild disease. Biomarker of type III collagen turnover (C3M/PROC3) showed higher levels in moderate and severe active disease in CD (1 ± 0.5 vs. 1.8 ± 1.1, p = 0.006). UC patients with moderate to severe activity showed higher levels of type III collagen turnover biomarker (C3M/PROC3) (1.1 ± 0.4 vs. 1.97 ± 1, p = 0.049). C4M correlated positively (r = 0.28, p = 0.03) and PRO-C3 correlated negatively (r = −0.29, p = 0.03) to SES-CD and combined in a multivariate regression model (r = 0.39, p = 0.009). C3M (r = 0.60, p = 0.004) and C4M (r = 0.36, p = 0.039) correlated positively with mMES.

Conclusion
In highly active CD, there is increased type IV collagen degradation and reduced type III collagen formation. Both biomarkers significantly correlate with SES-CD, with increased correlation when combined together. Type III collagen turnover is increased in highly active CD and UC. Biomarkers of types III and IV collagen degradation correlate significantly with mMES. Biomarkers of types III and IV collagen could be used to differentiate highly active CD from UC. Our findings suggest that biomarkers of basement membrane and interstitial matrix remodelling may be used as surrogate markers for monitoring of disease activity for UC and CD.