bluuebunny/crossref_metadata_2025_split huggingface dataset:
P011 New insights into the pathogenic potential and signalling network of NKG2D+ CD4+ T-cells in Crohn’s Disease
Authors: Modos D , Gul L , Lo J , Madgwick M , Cozzetto D , Lord G , Korcsmaros T , Powell N 
Published: January 2022

Abstract

Background
A subset of mucosal CD4+ T-cells expressing the NK cell receptor NKG2D (encoded by the KLRK1 gene) are enriched in patients with Crohn’s Disease (CD). In pre-clinical models of colitis, NKG2D expression was confined to a subset of colonic CD4+ T-cells that hyper-expressed interferon gamma. However, the functional role of these cells in mediating intestinal inflammation is unknown. Little is known about the transcriptional regulation of this population or the key signalling networks that they engage.

Methods
Pre-clinical models of CD, adoptive transfer experiments and in vitro culture systems were used to investigate the phenotype and pathogenicity of NKG2D using Klrk1-/- and WT mice. Signalling networks and interactions between NKG2D, key transcription factors and relevant cytokines were investigated by probing published single-cell RNA seq data (Martin et al. Cell 2019). Pathway members of potential interactions were collected from the literature, and the OmniPath database and we applied a heat propagation model.

Results
NKG2D-expressing T-cells were potently pathogenic when adoptively transferred to Rag1- mice, mediating more severe disease than Klrk1-/- CD4+ T-cells. Notably, Klrk-/- T-cells were still able to induce colitis, indicating that NKG2D-independent pathogenic pathways also exist.
Chromatin immunoprecipitation demonstrated that T-bet bound at the transcriptional start site of the Klrk1 locus in WT Th1 cells, but not Tbx21-/- Th1 T-cells. There were significantly fewer NKG2D expressing CD4+ T-cells from Tbx21-/- mice, and forced expression of T-bet in Tbx21-/-Ifng-/- double knockout CD4+ T-cells induced Klrk1 mRNA expression, consistent with T-bet being a transcriptional activator of the Klrk1 gene.
The NKG2D model network reached FOS, NFKB and JUN transcription factors depending on the presence of the NKG2D. The model distinguished two comparable pathway activation - one with active PRKCA in T-cells including CD8+ T-cells and one with less active PRKCA mostly in macrophages. Comparing the inflamed and uninflamed conditions, highly activated T-cells, and a population of Th17+ TRM cells had an active KLRK pathway only in inflamed Crohn disease.

Conclusion
NKG2D expressing colonic CD4+ T-cells are potent cytokine producing cells and are more colitigenic than their NKG2D-negative counterparts. Network-based heat propagation showed that specific populations of colonic memory T cells are active only in the inflamed condition, meanwhile, the model identified two types of cells that had active KLRK1 pathway: one which was PRKCA dependent activity and one which had non-PRKCA dependent activity. We also identify T-bet as an important transcriptional regulator of NKG2D expression.
P039 The role of α4β7+CLA+ ‘dual tropic’ T cells in inflammatory bowel disease
Authors: Gordon H , Rodger B , Hornsby E , McGuire J , Kok K B , Lindsay J O , Stagg A J 
Published: January 2022

Abstract

Background
Upon activation, T cells upregulate cell-surface molecules that enable entry into peripheral tissues. Retinoic acid (RA) produced by gut DC induces integrin α4β7 required for gut homing and represses CLA which facilitates skin entry; hence, these markers are usually reciprocally expressed. We aimed to identify the mechanisms enabling dual tropism, define the properties of α4β7+CLA+ T cells and compare blood and intestinal α4β7+CLA+ T cells in health and IBD.

Methods
Using flow cytometry, expression of α4β7 and CLA was assessed on (i) ex vivo human T cells from blood and intestinal tissue and; (ii) naïve CD4 T cells stimulated in vitro with antibodies (anti-CD3/28/2), or allogeneic DCs. The role of RA signalling and p38MAPK in CLA expression were investigated with respective antagonists Ro41-5253 and SB203580. Transwell studies were used to investigate whether DC-T cell direct contact was required for CLA expression. Single cell RNA sequencing (scRNAseq) of in vitro activated T cells was undertaken, with feature barcode antibody labelling of β7 and CLA. Differential gene expression and pathway analysis was undertaken.

Results
DC-activated T cells contained both α4β7+ and CLA+ cells and included a dual tropic population co-expressing both markers. In contrast, antibody-activated T cells expressed α4β7 but did not express CLA unless RA receptor alpha (RARa) signalling was also inhibited. The DC-derived signals that allowed CLA expression were partially dependent on p38MAPK and required DC-T cell contact for optimal activity. A high frequency of dual tropic T cells produced IFNg or TNFa.
ScRNAseq identified 270 genes differentially expressed (fold change &lt;-2 or &gt;+2, FDR&lt;0.01) between α4β7+CLA+ and α4β7+CLA- T populations generated by DC stimulation. Expression of FUT7, a fucosyltransferase implicated in generation of the CLA moiety, was increased in the α4β7+CLA+ population and overall 27 pathways were significantly enriched (p&lt;0.05) in this population, including intestinal immune networks and IBD.
Ex vivo, CD4+ and CD8+ dual tropic T cells were found in peripheral blood and intestinal tissue. CD4+ dual tropic T cells were significantly increased in the colon of patients with IBD (p=0.03).

Conclusion
DC-derived signals acting via p38MAPK generate pro-inflammatory dual tropic T cells even in the presence of active RA signalling. CLA expression may act as an additional ligand in gut trafficking by binding to selectins on the inflamed vascular endothelium and has a potential role in both gut and skin inflammation in IBD.
P041 Characterization of cytokine and drug concentrations in serum, mucosa and faeces during induction treatment of moderate-to-severe ulcerative colitis with anti-TNF monoclonal antibodies
Authors: Van Oostrom J , Hanzel J , Verstockt B , Singh S , Smith J , Stylli J , Vermeire S , D’Haens G 
Published: January 2022

Abstract

Background
It remains unclear why up to 30% of patients with moderate-to-severe ulcerative colitis (UC) do not respond to anti-TNF treatment (anti-TNF). In this study, we explored multi compartment cytokine and anti-TNF concentrations in peripheral blood (PB), faeces (F) and colonic mucosa (CM) before, during and after anti-TNF induction treatment in UC.

Methods
91 UC patients starting infliximab or adalimumab were included at two tertiary centres (table 1). Patients underwent clinical and endoscopic assessment with CM, PB, and F sampling at baseline. Follow-up CM samples were taken at first follow-up endoscopy during the first year. Follow-up PB and F samples were taken at follow-up endoscopy and optionally on day 3, 7, 14, and 28.
Response (R) was defined as Mayo endoscopic score (MES) 0–1 (assessed by a blinded independent reader) and subsequent anti-TNF therapy for &gt; 1 year. All other patients were classified as non-responders (NR).
TNFα, IL6, IL10 and drug levels were measured in all compartments and OSM and IL12p40 in PB and CM (F anti-TNF with ELISA and cytokines and PB and CM anti-TNF with a bead-based immunoassay). CM samples were normalised for gram of protein (gop).
To differentiate pharmacokinetic (PK) from pharmacodynamic (PD) failures, we calculated CM anti-TNF/ TNFα. We classified values above the median as sufficient PK (Good PK) and values below as insufficient PK (Bad PK).

Results
We analysed CM, PB and F samples from 71, 91 and 32 patients respectively. Baseline cytokine concentrations were similar in all compartments.
At follow-up (table 2), PB IL6 was lower in R (p=0.045). All F cytokines were significantly higher in R (TNFα: p&lt;0.001; IL6: p&lt;0.001; IL10: p&lt;0.001). In CM, IL6 and OSM were lower and IL12p40 higher in R (IL6: p=0.004; OSM: p=0.01; IL12p40: p=0.021).
After stratification of CM samples for PK readouts (table 3, figure 1), IL6 was highest in Good PK NR vs Good PK R (6905 vs 2980 ng/gop, p = 0.002) and OSM was lowest in R vs NR groups (2466 vs 36111 ng/gop in Good PK R vs Good PK NR, p = 0.002). Trends were observed towards highest IL10 in Bad PK R and Highest IL12p40 in Good PK R, although not significant.

Conclusion
Baseline cytokine concentrations in PB, CM and F did not predict response to anti-TNF induction treatment in UC. At follow-up, response (R) was characterised by lower PB IL6; higher F cytokines; lower CM IL6 and OSM and higher IL12p40.
After stratification for PK, pharmacodynamic failure (Good PK NR) was characterised by highest CM IL6, indicating it is a driver to inflammation alternative to anti-TNF. R was characterised by lowest OSM regardless of PK, indicating OSM mainly reflects disease activity. Further research focuses on cytokine-PK interplay contributing to anti-TNF (non-) response.
P067 Human intestinal CD103+SIRPα+ conventional dendritic cells from patients with Crohn′s disease prime IL-17+ T-cells
Authors: Bernardo D , Arribas-Rodriguez E , De Castro C G , Fiz-Lopez A , De Prado A , Fernandez-Salazar L , Barrio J , Izquierdo S , Garcia-Alonso J , De Andres B , Garcia-Abril J M , Romero A , Sanchez-Gonzalez J , Garrote J A , Arranz E 
Published: January 2022

Abstract

Background
Crohn’s disease (CD) is a chronic inflammation of the human gastrointestinal tract. Intestinal conventional dendritic cells (cDC) maintain the balance between immunity against pathogens and tolerance towards nutrients and commensals. However, there is not much information about DC composition, phenotype and function in the human gut in CD.

Methods
Human intestinal resections were obtained from patients with ileocolonic CD and proximal colonic cancer. Both inflamed and non-inflamed tissue was obtained from the ileum and colon from CD patients. The non-affected tissue from the proximal (terminal ileum) and distal (proximal colon) sides of the colonic cancer resections (minimum 10cm distance from the tumor) constituted the reference tissue.
Tissue was disaggregated and cDC and macrophages enriched by flow-sorting. Human intestinal cDC were identified within singlet viable leukocytes as CD14-CD64-HLA-DR+CD11c+. Type 1 cDC (cDC1) were defined as CD103+SIRPα- while type 2 cDC (cDC2) and identified as SIRPα+ and further divided into subsets based on the expression of CD103. Macrophages were identified within singlet viable leukocytes as CD14+CD64+HLA-DR+. All three cDC subsets and total macrophages were used to stimulate allogeneic CD4+ naïve T-cells which were further characterized by flow cytometry.

Results
All 3 human intestinal cDC subsets from non-CD controls were able to stimulate naïve T-cells as opposed to intestinal macrophages. Indeed, stimulated T-cells produced IL-10 and co-expressed FoxP3, being this capacity increased in both CD103+ subsets (CD103+SIRPα- and CD103+SIRPα+). Moreover, all three ileal cDC subsets stimulated T-cells more efficiently than their paired colonic counterparts, although the acquired T-cell profile did not differ among tissues. None of the cDC subets, on the contrary, induced the expression of IFNγ or IL-17 on stimulated T-cells. Referred to CD patients, all three colonic cDC subsets were more stimulatory than their counterparts from the control tissue, while CD103+SIRPα+ cDC from the inflamed ileum primed the generation of IL-17+ responding T-cells.

Conclusion
All human intestinal cDC subsets prime the generation of regulatory FoxP3+ and IL-10+ T-cells. This capacity was increased in both CD103+SIRPα- and CD103+SIRPα+, in agreement with the regulatory functions attributed to CD103+ cDC. All ileal cDC subsets were also more stimulatory than their colonic counterparts. Nevertheless, colonic cDC from CD patients were more stimulatory that non-CD controls, rendering CD103+SIRPα+ cDC from the inflamed tissue in CD with a higher capacity to generate Th17 cells, which suggests that their regulatory functions can be reshaped by the inflamed tissue microenvironment.
P069 Design and delivery of anti-oxidant inorganic complexes mimicking the superoxide dismutase for inflammatory bowel diseases treatment
Authors: Schanne G , Zoumpoulaki M , Demignot S , Delsuc N , Policar C , Seksik P 
Published: January 2022

Abstract

Background
Oxidative stress (OS), defined by an increased flux of reactive oxygen species (ROS), plays an important role in the pathogenesis, the perpetuation and the amplification of inflammatory bowel diseases. In normal state, enzymatic defenses regulate the concentration of ROS but in IBD, some of them, including the superoxide dismutase (SOD), are weakened. The use of molecules mimicking the SOD activity may compensate for this deficiency and reduce the OS. 4 molecules were characterized for their SOD activity, their stability and their inertness. Their anti-inflammatory activity were assessed in a cellular model of epithelial inflammation and in a murine model of colitis and correlated to their physico-chemical properties.

Methods
The studied SOD mimics display similar structures differing by the presence of a propyl and a cyclohexyle group aiming to improve their kinetic inertness. To generate an inflammation associated to ROS production, intestinal epithelial (HT29) cells were stably transfected to over-express MD2, a co-receptor for LPS binding to TLR-4 receptor. The cells were stimulated by LPS (0.1µg/mL) and incubated with the SOD mimics (6 hours, 0,1-100µM). The secretion of IL-8 was measured by ELISA and the MnSOD expression by Western Blot. The SOD mimics were administrated to a murine model of colitis chemically-induced by DNBS. To protect them from the gastric acidity, the mimics were encapsulated in delivery vector. The monitoring of weight and the determination of intestinal permeability and macroscopic scores were performed.

Results
The mimics containing a cyclohexyle group display kinetic rates for metal exchanges that are up to 30-fold lower meaning that they are more inert. At 100µM, the four SOD mimics induced a significative decrease in IL-8 secretion (&lt;35%/positive control (PT), p&lt;0,02) and in MnSOD expression (&lt;20%/PT, p&lt;0,033). At 10µM, only the mimics containing the cyclohexyle group are still able to significatively counteract the LPS-induced secretion in IL-8 (&lt;30/PT, p&lt;0,001) and over-expression of MnSOD (20%/PT, p&lt;0,02). Their bioactivity at lower doses can be correlated to their higher inertness. Concerning in vivo activity, preliminary assays show a faster recovery of the mice, that have received the SOD mimics with the cyclohexyle group. 3 days following DNBS injection, the mice of this group recovered in average 89% of their starting weight while the control mice recovered only 83%.

Conclusion
The SOD mimics display anti-inflammatory and antioxidant activity on HT29-MD2 cells, which is dependent on their inertness. Assays on a murine model of colitis seems to follow the same trend. The use of antioxidant treatments based on SOD mimics may reduce the inflammation rising from oxidative stress in IBD.
P116 Screening for low BMI and unintentional weight loss alone is insufficient to identify patients at risk for malnutrition in an outpatient IBD population
Authors: Hendrix E M B , Spooren C E G M , Grommen D , Mujagic Z , Pierik M J , Jonkers D M A E 
Published: January 2022

Abstract

Background
Inflammatory bowel disease (IBD) is associated with malnutrition, which can further impair disease course and quality of life. Therefore, guidelines advocate screening of patients in clinical practice. The prevalence of malnutrition in IBD-cohorts however, varies widely, mainly due to differences in parameters used. The primary aim of the present study was to assess the prevalence of malnutrition using single and a combined set of parameters (Global Leadership Initiative on Malnutrition (GLIM) criteria). Secondary aims were i) to evaluate the accuracy of screening recommendations given in current IBD guidelines and ii) to explore which patients have an increased risk of malnutrition.

Methods
Malnutrition was defined by the GLIM criteria, based on the combination of a phenotypic (i.e. non-volitional weight loss, low body mass index (BMI), or reduced muscle mass) and an etiologic criterium (i.e. reduced food intake or assimilation, and disease burden or inflammation). Malnutrition was also determined using single parameters for impaired body composition, muscle strength or caloric intake (Table 1), and the combination of low BMI and unintentional weight loss as advised in current IBD guidelines. To screen for malnutrition, the Short Nutritional Assessment Questionnaire (SNAQ) and Malnutrition Universal Screening Tool (MUST) were completed. Independent risk factors (i.e. clinical and demographic factors) for malnutrition were analyzed by multivariable logistic regression.

Results
Of the 200 included patients (139 CD, 61 UC), 41 (20.5%) fulfilled the GLIM criteria, 95 (47.5%) had at least one parameter for malnutrition impaired (Figure 1). The fat free mass index was most often affected. When unintentional weight loss and/or low BMI was used as screening marker for nutritional status in line with current IBD guidelines, 29 (14.5%) patients would have been identified (Figure 2). Screening for malnutrition using the SNAQ and MUST detected 44 (22.0%) and 23 (12.9%) patients with a positive score. Only female sex was associated with malnutrition when at least one parameter was impaired (OR 2.47, 95% CI 1.35–4.51).

Conclusion
Malnutrition prevalence among IBD outpatients according to the GLIM criteria was found to be 20.5%. Almost half of the IBD outpatients had malnutrition as defined by various single parameters and irrespective of disease characteristics. Screening instruments and/or markers according to current IBD guidelines, did not identify a substantial part of the patients. Therefore, screening for malnutrition is recommended for all IBD outpatients by multiple parameters, with special attention for assessing fat free mass and reduced intake.
P125 Ankylosing spondylitis can influence the outcome of inflammatory bowel disease
Authors: Jun Y K , Kim A H , Park J , Choi E , Yoon H , Kang H W , Lee H J , Im J P , Kim J S , Seong-Joon K 
Published: January 2022

Abstract

Background
Both inflammatory bowel disease (IBD) and ankylosing spondylitis (AS) are inflammatory diseases but there was little previous study that demonstrate the IBD severity and outcome in IBD patients with concomitant AS.

Methods
Patients’ records were collected from 3 tertiary hospitals (Seoul national university hospital, Seoul national university Bundang hospital, and Seoul metropolitan government Seoul national university Boramae medical center) from October 2004 to June 2021. Patients with IBD and concomitant AS (IBD-AS group) were identified and propensity score matching (PSM) was applied to match the IBD-AS group and IBD patients without AS (only IBD group).

Results
After PSM, significantly more patients in IBD-AS group had colectomies (p = .017) or were prescribed biologics (p &lt; .001), immunosuppressive agents (p = .021), and steroid (p = .017) than in only IBD group. The number of patients treated with biologics (p &lt; .001) or immunosuppressive agents (p = .032) were significantly greater in UC-AS group than in only UC group. There was no significant difference in outcomes between only CD group and CD-AS group. In logistic regression analyses, identified that concomitant AS was a significant factor associated with biologics treatment in patients with IBD. Kaplan-Meier analyses demonstrated that there was a significant difference in the probability of starting biologics treatment between IBD patients with and without concomitant AS (p = 0.002). In UC patients, the probability of starting biologics was also significantly different according to concomitance of AS (p &lt; 0.001). Concomitant AS was a risk factor for predicting biologics treatment in patients with UC in Cox regression analysis after adjustment UC (adjusted hazard ratio, 6.296; confidence interval, 2.243 to 17.668; p &lt; 0.001)

Conclusion
Patients with IBD-AS group were more likely to have a higher severity than in only IBD group. The current study result can help IBD specialists understand and treat patients with IBD and concomitant AS better.
P133 Point-of-care measurement of infliximab and adalimumab trough levels by the fluorescence resonance energy transfer (FRET) assay
Authors: Cerna K , Kastylova K , Sebek P , Luxova S , Lukas M 
Published: January 2022

Abstract

Background
Therapeutic drug monitoring (TDM) is the clinical practice of measuring serum drug concentrations to guide clinical decision-making. In IBD patients, it is applied mainly in infliximab (IFX) and adalimumab (ADL) therapy. TDM is either performed reactively in response to disease activity, or proactively in the absence of symptoms. The aim of the present study was to compare two analytical methods for the detection of trough infliximab and adalimumab levels (TL): point-of-care (POCT) fluorescence resonance energy transfer (FRET) method, and the enzyme- linked immunosorbent immunoassay (ELISA).

Methods
TL IFX were measured in n=24 and TL ADL in n= 21 blood samples. For ELISA, standard peripheral venous samples were used, and for POCT FRET measurements, capillary finger puncture was performed. Two assay results were compared evaluating the regression relationship using Spearman’s correlation coefficient and the Bland-Altman plots. Cohen’s kappa analysis was used to compare the degree of qualitative agreement between the two data sets. Repeatability was assessed by the difference of the repeated measurements, their means and standard deviations.

Results
The high degree of correlation was found between the measured values, both qualitative and quantitative (p &lt; 0.0001 for both IFX and ADL), either expressed by the Spearman’s correlations, or by Bland-Altman analysis. Cohen’s weighted kappa 0.903 for IFX and 0.834 for ADL showed substantial/almost perfect agreement of qualitative results. Closeness of the agreement of successive measurements was very acceptable for both TLs of both drugs.

Conclusion
Point-of-care detection of IFX and ADL trough levels by FRET is accurate, comparable to the reference method, analysis is sufficiently consistent. For clinical sites with a proactive approach to TDM, FRET may be a useful tool for decision-making regarding individualization of biologic therapy.
Acknowledgements: We thank ProciseDX company for the ProciseDX analyzer rental, and for providing the laboratory diagnostics.