439 results on '"Thomas T. MacDonald"'
Search Results
2. Preclinical development of a bispecific TNFα/IL-23 neutralising domain antibody as a novel oral treatment for inflammatory bowel disease
- Author
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Kevin J. Roberts, Marion F. Cubitt, Timothy M. Carlton, Lurdes Rodrigues-Duarte, Luana Maggiore, Ray Chai, Simon Clare, Katherine Harcourt, Thomas T. MacDonald, Keith P. Ray, Anna Vossenkämper, Michael R. West, and J. Scott Crowe
- Subjects
Medicine ,Science - Abstract
Abstract Anti-TNFα and anti-IL-23 antibodies are highly effective therapies for Crohn’s disease or ulcerative colitis in a proportion of patients. V56B2 is a novel bispecific domain antibody in which a llama-derived IL-23p19-specific domain antibody, humanised and engineered for intestinal protease resistance, V900, was combined with a previously-described TNFα-specific domain antibody, V565. V56B2 contains a central protease-labile linker to create a single molecule for oral administration. Incubation of V56B2 with trypsin or human faecal supernatant resulted in a complete separation of the V565 and V900 monomers without loss of neutralising potency. Following oral administration of V900 and V565 in mice, high levels of each domain antibody were detected in the faeces, demonstrating stability in the intestinal milieu. In ex vivo cultures of colonic biopsies from IBD patients, treatment with V565 or V900 inhibited tissue phosphoprotein levels and with a combination of the two, inhibition was even greater. These results support further development of V56B2 as an oral therapy for IBD with improved safety and efficacy in a greater proportion of patients as well as greater convenience for patients compared with traditional monoclonal antibody therapies.
- Published
- 2021
- Full Text
- View/download PDF
3. Aryl Hydrocarbon Receptor Interacting Protein Maintains Germinal Center B Cells through Suppression of BCL6 Degradation
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Dijue Sun, Urszula Stopka-Farooqui, Sayka Barry, Ezra Aksoy, Gregory Parsonage, Anna Vossenkämper, Melania Capasso, Xinyu Wan, Sherine Norris, Jennifer L. Marshall, Andrew Clear, John Gribben, Thomas T. MacDonald, Christopher D. Buckley, Márta Korbonits, and Oliver Haworth
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Biology (General) ,QH301-705.5 - Abstract
Summary: B cell lymphoma-6 (BCL6) is highly expressed in germinal center B cells, but how its expression is maintained is still not completely clear. Aryl hydrocarbon receptor interacting protein (AIP) is a co-chaperone of heat shock protein 90. Deletion of Aip in B cells decreased BCL6 expression, reducing germinal center B cells and diminishing adaptive immune responses. AIP was required for optimal AKT signaling in response to B cell receptor stimulation, and AIP protected BCL6 from ubiquitin-mediated proteasomal degradation by the E3-ubiquitin ligase FBXO11 by binding to the deubiquitinase UCHL1, thus helping to maintain the expression of BCL6. AIP was highly expressed in primary diffuse large B cell lymphomas compared to healthy tissue and other tumors. Our findings describe AIP as a positive regulator of BCL6 expression with implications for the pathobiology of diffuse large B cell lymphoma. : BCL6 overexpression contributes to the pathobiology of diffuse large B cell lymphoma (DLBCL). Sun et al. find that the co-chaperone aryl hydrocarbon receptor interacting protein (AIP), whose high expression is associated with reduced survival of DLBCL patients, helps maintain BCL6 expression by facilitating the removal of ubiquitin from BCL6. Keywords: AIP, BCL6, FBXO11, UCHL1, ubiquitination, lymphoma
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- 2019
- Full Text
- View/download PDF
4. The Role of Cytokines in the Fibrotic Responses in Crohn’s Disease
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Renata Curciarello, Guillermo H. Docena, and Thomas T. MacDonald
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Crohn’s disease ,fibrosis ,fibrostenosis ,cytokines ,strictures ,extracellular matrix ,Medicine (General) ,R5-920 - Abstract
Crohn’s disease is an idiopathic disorder of the gut thought to be caused by a combination of environmental and genetic factors in susceptible individuals. It is characterized by chronic transmural inflammation of the terminal ileum and colon, with typical transmural lesions. Complications, including fibrosis, mean that between 40 and 70% of patients require surgery in the first 10 years after diagnosis. Presently, there is no evidence that the current therapies which dampen inflammation modulate or reverse intestinal fibrosis. In this review, we focus on cytokines that may lead to fibrosis and stenosis and the contribution of experimental models for understanding and treatment of gut fibrosis.
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- 2017
- Full Text
- View/download PDF
5. Supplementary Figures 1-4, Methods, Table 1 from Smad7 Expression in T cells Prevents Colitis-Associated Cancer
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Massimo C. Fantini, Giovanni Monteleone, Francesco Pallone, Thomas T. Macdonald, Markus F. Neurath, Christoph Becker, Daniele Fina, Massimiliano Sarra, Carmine Stolfi, Maximilian J. Waldner, and Angelamaria Rizzo
- Abstract
PDF file - 630K
- Published
- 2023
6. Data from Smad7 Expression in T cells Prevents Colitis-Associated Cancer
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Massimo C. Fantini, Giovanni Monteleone, Francesco Pallone, Thomas T. Macdonald, Markus F. Neurath, Christoph Becker, Daniele Fina, Massimiliano Sarra, Carmine Stolfi, Maximilian J. Waldner, and Angelamaria Rizzo
- Abstract
Patients with inflammatory bowel disease (IBD) have an increased risk of developing colorectal cancer due to chronic inflammation. In IBD, chronic inflammation relies upon a TGFβ signaling blockade, but its precise mechanistic relationship to colitis-associated colorectal cancer (CAC) remains unclear. In this study, we investigated the role of the TGFβ signaling inhibitor Smad7 in CAC pathogenesis. In human colonic specimens, Smad7 was downregulated in CD4+ T cells located in the lamina propria of patients with complicated IBD compared with uncomplicated IBD. Therefore, we assessed CAC susceptibility in a transgenic mouse model where Smad7 was overexpressed specifically in T cells. In this model, Smad7 overexpression increased colitis severity, but the mice nevertheless developed fewer tumors than nontransgenic mice. Protection was associated with increased expression of IFNγ and increased accumulation of cytotoxic CD8+ and natural killer T cells in the tumors and peritumoral areas. Moreover, genetic deficiency in IFNγ abolished the Smad7-dependent protection against CAC. Taken together, our findings defined a novel and unexpected role for Smad7 in promoting a heightened inflammatory response that protects against CAC. Cancer Res; 71(24); 7423–32. ©2011 AACR.
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- 2023
7. Spatiotemporal regulation of galectin-1-induced T-cell death in lamina propria from Crohn’s disease and ulcerative colitis patients
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Gabriel A. Rabinovich, Karina Mariño, Cecilia Isabel Muglia, Renata Curciarello, Gustavo Correa, Paolo Biancheri, Martin Yantorno, Antonio Di Sabatino, Alicia M. Sambuelli, Thomas T. MacDonald, Marta A. Toscano, Luciano Gastón Morosi, Guillermo Horacio Docena, Rodrigo Papa-Gobbi, and Andrés Rocca
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0301 basic medicine ,Peanut agglutinin ,Cancer Research ,Clinical Biochemistry ,Pharmaceutical Science ,Inflammation ,Inflammatory bowel disease ,03 medical and health sciences ,0302 clinical medicine ,medicine ,Pharmacology ,Lamina propria ,Crohn's disease ,biology ,business.industry ,Biochemistry (medical) ,Cell Biology ,medicine.disease ,Ulcerative colitis ,030104 developmental biology ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Immunology ,biology.protein ,Immunohistochemistry ,medicine.symptom ,business ,CD8 - Abstract
Inflammatory bowel disease (IBD), including Crohn’s disease (CD) and ulcerative colitis (UC), is characterized by chronic, relapsing intestinal inflammation. Galectin-1 (Gal-1) is an endogenous lectin with key pro-resolving roles, including induction of T-cell apoptosis and secretion of immunosuppressive cytokines. Despite considerable progress, the relevance of Gal-1-induced T-cell death in inflamed tissue from human IBD patients has not been ascertained. Intestinal biopsies and surgical specimens from control patients (n = 52) and patients with active or inactive IBD (n = 97) were studied. Gal-1 expression was studied by RT-qPCR, immunoblotting, ELISA and immunohistochemistry. Gal-1-specific ligands and Gal-1-induced apoptosis of lamina propria (LP) T-cells were determined by TUNEL and flow cytometry. We found a transient expression of asialo core 1-O-glycans in LP T-cells from inflamed areas (p
- Published
- 2021
8. Human Neutrophil Elastase Proteolytic Activity in Ulcerative Colitis Favors the Loss of Function of Therapeutic Monoclonal Antibodies
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Samantha Jones, Antonio Di Sabatino, Thomas T. MacDonald, Renata Curciarello, Toni Sobande, Klaartje Kok, Paolo Giuffrida, and Guillermo Horacio Docena
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0301 basic medicine ,Proteases ,biology ,medicine.drug_class ,business.industry ,Immunology ,Elastase ,Monoclonal antibody ,medicine.disease ,Inflammatory bowel disease ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,Intestinal mucosa ,030220 oncology & carcinogenesis ,Neutrophil elastase ,biology.protein ,medicine ,Immunology and Allergy ,Tumor necrosis factor alpha ,business ,Elafin - Abstract
Purpose Proteases play an essential role in the pathophysiology of inflammatory bowel disease (IBD), contributing to the intestinal mucosal lesions through the degradation of the extracellular matrix and alteration of the barrier function. Ulcerative colitis (UC) is characterized by an extensive infiltrate of neutrophils into the mucosa and hence, increased proteolytic activity. Human neutrophil elastase (HNE) is a serine protease that has been reported to be increased in UC patients' intestinal mucosa. Based on our previous studies, we hypothesized that HNE might induce proteolytic degradation and loss of function of therapeutic monoclonal antibodies in IBD patients. Patients and Methods Elastase expression and elastinolytic activity were determined in mucosal explants from ulcerative colitis patients (n=6) and cultured ex vivo in the presence or absence of recombinant elafin. Enzymatic digestions of therapeutic monoclonal antibodies were performed using recombinant HNE and elafin. The integrity of the therapeutic antibodies was evaluated by immunoblotting and protein G binding assay, whereas their TNF-neutralizing activity was assessed with a reporter cell line. Results We found that HNE and its elastinolytic activity were increased in the gut mucosa of UC patients. We also demonstrated that HNE cleaved biological drugs, impairing the TNF-α neutralizing capacity of anti-TNF monoclonal antibodies. This proteolytic degradation was inhibited by the addition of the specific inhibitor, elafin. Conclusion Our results suggest that the high level of proteolytic degradation by mucosal neutrophil elastase, along with a potential imbalance with elafin, contributes to the loss of function of biologic agents, which are currently used in patients with IBD. These findings might explain the non-responsiveness of UC patients to therapeutic monoclonal antibodies and suggest the potential beneficial concomitant use of elafin in this treatment.
- Published
- 2020
9. Extended pharmacodynamic responses observed upon PROTAC-mediated degradation of RIPK2
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Jenni Cryan, Don T. Fisher, Michael Reilly, Ian Edward David Smith, Anna Vossenkämper, Gillian F. Watt, Ian Churcher, John D. Harling, Allison M. Beal, Thomas T. MacDonald, Alina Mares, Phoebe Dace, Bartholomew J. Votta, Marcus Bantscheff, Jane Denyer, Paul Scott-Stevens, Afjal Hussain Miah, Andrew B. Benowitz, Aditya R. Thawani, Carol A. Capriotti, Mark David Rackham, Nico Zinn, and Pamela A. Haile
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Male ,THP-1 Cells ,Anti-Inflammatory Agents ,Medicine (miscellaneous) ,Medicinal chemistry ,Pharmacology ,01 natural sciences ,Serine ,Rats, Sprague-Dawley ,Tissue Culture Techniques ,Ubiquitin ,Crohn Disease ,Enzyme Stability ,lcsh:QH301-705.5 ,0303 health sciences ,biology ,medicine.diagnostic_test ,Drug discovery ,Chemistry ,Ubiquitin ligase ,Injections, Intravenous ,Cytokines ,Female ,medicine.symptom ,Inflammation Mediators ,General Agricultural and Biological Sciences ,Proteasome Endopeptidase Complex ,Proteolysis ,Ubiquitin-Protein Ligases ,General Biochemistry, Genetics and Molecular Biology ,Article ,RIPK2 ,03 medical and health sciences ,Receptor-Interacting Protein Serine-Threonine Kinase 2 ,medicine ,Animals ,Humans ,Pharmacokinetics ,Rats, Wistar ,Protein kinase A ,030304 developmental biology ,Inflammation ,Dose-Response Relationship, Drug ,010405 organic chemistry ,Ubiquitination ,0104 chemical sciences ,Mechanism of action ,Pharmacodynamics ,lcsh:Biology (General) ,Drug Design ,biology.protein ,Leukocytes, Mononuclear ,Colitis, Ulcerative - Abstract
Proteolysis-Targeting Chimeras (PROTACs) are heterobifunctional small-molecules that can promote the rapid and selective proteasome-mediated degradation of intracellular proteins through the recruitment of E3 ligase complexes to non-native protein substrates. The catalytic mechanism of action of PROTACs represents an exciting new modality in drug discovery that offers several potential advantages over traditional small-molecule inhibitors, including the potential to deliver pharmacodynamic (PD) efficacy which extends beyond the detectable pharmacokinetic (PK) presence of the PROTAC, driven by the synthesis rate of the protein. Herein we report the identification and development of PROTACs that selectively degrade Receptor-Interacting Serine/Threonine Protein Kinase 2 (RIPK2) and demonstrate in vivo degradation of endogenous RIPK2 in rats at low doses and extended PD that persists in the absence of detectable compound. This disconnect between PK and PD, when coupled with low nanomolar potency, offers the potential for low human doses and infrequent dosing regimens with PROTAC medicines., Mares et al. develop Proteolysis-Targeting Chimeras (PROTACs) that degrade its target RIPK2 in vivo at low doses for a prolonged period of time. This study suggests that PROTAC has a therapeutic potential that is superior to traditional RIPK2 small-molecule inhibitors.
- Published
- 2020
10. Comparison of cytokine and phosphoprotein profiles in idiopathic and Crohn’s disease-related perianal fistula
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Thomas T. MacDonald, Charles H. Knowles, Omar Musbahi, and James B Haddow
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Crohn’s disease ,Anal fistula ,medicine.medical_specialty ,Crohn's disease ,business.industry ,medicine.medical_treatment ,Observational Study ,Pathogenesis ,Phosphoproteins ,medicine.disease ,Gastroenterology ,digestive system diseases ,03 medical and health sciences ,0302 clinical medicine ,Cytokine ,Perianal fistula ,030220 oncology & carcinogenesis ,Internal medicine ,Phosphoprotein ,Cytokines ,Medicine ,030211 gastroenterology & hepatology ,business - Abstract
BACKGROUND Perianal fistulae are either primary (idiopathic) or secondary [commonly associated with Crohn’s disease, (CD)]. It is assumed, although not proven, that the pathophysiology differs. AIM To systematically compare the clinical phenotypes, cytokine and phosphoprotein profiles of idiopathic and CD-related perianal fistulae. METHODS Sixty-one patients undergoing surgery for perianal fistula were prospectively recruited (48 idiopathic, 13 CD) into a cohort study. Clinical data, including the Perineal Disease Activity Index (PDAI) and EQ-5D-5L were collected. Biopsies of the fistula tract, granulation tissue, internal opening mucosa and rectal mucosa were obtained at surgery. Concentrations of 30 cytokines and 39 phosphoproteins were measured in each biopsy using a magnetic bead multiplexing instrument and a chemiluminescent antibody array respectively. Over 12000 clinical and 23500 laboratory measurements were made. RESULTS The PDAI was significantly higher (indicating more active disease) in the CD group with a mean difference of 2.40 (95%CI: 0.52-4.28, P = 0.01). Complex pathoanatomy was more prevalent in the CD group, namely more multiple fistulae, supralevator extensions, collections and rectal thickening. The IL-12p70 concentration at the internal opening specimen site was significantly higher (median difference 19.7 pg/mL, 99%CI: 0.2-40.4, P = 0.008) and the IL-1RA/IL-1β ratio was significantly lower in the CD group at the internal opening specimen site (median difference 15.0, 99%CI = 0.4-50.5, P = 0.008). However in the remaining 27 cytokines and all 39 of the phosphoproteins across the four biopsy sites, no significant differences were found between the groups. CONCLUSION CD-related perianal fistulae are more clinically severe and anatomically complex than idiopathic perianal fistulae. However, overall there are no major differences in cytokine and phosphoprotein profiles.
- Published
- 2019
11. Preclinical development of a bispecific TNFα/IL-23 neutralising domain antibody as a novel oral treatment for inflammatory bowel disease
- Author
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Katherine Harcourt, Michael R. West, Thomas T. MacDonald, Lurdes Rodrigues-Duarte, Anna Vossenkämper, Keith P. Ray, Ray Chai, Luana Maggiore, Marion F. Cubitt, Timothy M. Carlton, Simon Clare, J. Scott Crowe, Kevin J. Roberts, and Apollo - University of Cambridge Repository
- Subjects
medicine.drug_class ,Science ,Drug Evaluation, Preclinical ,Monoclonal antibody ,Interleukin-23 ,Inflammatory bowel disease ,Article ,Mice ,Oral administration ,Antibodies, Bispecific ,medicine ,Antibody fragment therapy ,Animals ,Humans ,Potency ,Crohn's disease ,Multidisciplinary ,biology ,Tumor Necrosis Factor-alpha ,business.industry ,Antibodies, Monoclonal ,Inflammatory Bowel Diseases ,medicine.disease ,Ulcerative colitis ,Mice, Inbred C57BL ,Immunology ,biology.protein ,Medicine ,Female ,Tumor necrosis factor alpha ,Antibody therapy ,Antibody ,business - Abstract
Anti-TNFα and anti-IL-23 antibodies are highly effective therapies for Crohn’s disease or ulcerative colitis in a proportion of patients. V56B2 is a novel bispecific domain antibody in which a llama-derived IL-23p19-specific domain antibody, humanised and engineered for intestinal protease resistance, V900, was combined with a previously-described TNFα-specific domain antibody, V565. V56B2 contains a central protease-labile linker to create a single molecule for oral administration. Incubation of V56B2 with trypsin or human faecal supernatant resulted in a complete separation of the V565 and V900 monomers without loss of neutralising potency. Following oral administration of V900 and V565 in mice, high levels of each domain antibody were detected in the faeces, demonstrating stability in the intestinal milieu. In ex vivo cultures of colonic biopsies from IBD patients, treatment with V565 or V900 inhibited tissue phosphoprotein levels and with a combination of the two, inhibition was even greater. These results support further development of V56B2 as an oral therapy for IBD with improved safety and efficacy in a greater proportion of patients as well as greater convenience for patients compared with traditional monoclonal antibody therapies.
- Published
- 2021
12. An Evaluation of Alveolar Bone Grafting in the UK and Ireland
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Norman Hay, Thomas T. MacDonald, Preeti Jauhar, and Brijesh Patel
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business.industry ,Radiography ,Grafting (decision trees) ,medicine.medical_treatment ,Dentistry ,030206 dentistry ,030230 surgery ,Bone grafting ,03 medical and health sciences ,0302 clinical medicine ,Otorhinolaryngology ,Medicine ,Oral Surgery ,Alveolar bone grafting ,business - Abstract
Objective: To determine the main factors influencing the timing of alveolar bone grafting among cleft teams in the UK and Ireland, to assess the types of radiographs used to evaluate bone grafting sites pre- and postoperatively and the views of the profession on orthodontic expansion prior to grafting. Design: An online survey consisting of 24 questions was compiled and emailed to 53 orthodontists and surgeons in all 12 Cleft Hub Units in the UK and Ireland. Results: All units in the UK and Ireland responded with 51 responses, 39 complete and 12 partial responses, obtained from cleft surgeons and orthodontists. The majority of units are using dental criteria (75%) as a guide to timing alveolar bone grafting. Most units take a postoperative radiograph at 6 months but the view being taken varied. When asked if four cases were ready for grafting based on their radiographs, there was clear agreement by a significant majority for 3 cases but for 1 only a minimal majority (61%). The most common donor site chosen for the graft is the Iliac crest (92.9%). There was excellent agreement for 2 cases asking when to use expansion but poor agreement for one, 55% saying they would expand and 45% saying no. Conclusion: Overall there is good agreement among cleft teams in the UK and Ireland about management of alveolar bone grafting; however, a clearer consensus on preoperative orthodontic expansion may be needed.
- Published
- 2021
13. Oral Anti-Tumour Necrosis Factor Domain Antibody V565 Provides High Intestinal Concentrations, and Reduces Markers of Inflammation in Ulcerative Colitis Patients
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Keith P. Ray, Gareth Parkes, Suhail Nurbhai, Michael R. West, Luana Maggiore, Hafeez Mohammed, Timothy M. Carlton, J. Scott Crowe, Jill Reckless, Thomas T. MacDonald, Kevin J. Roberts, Marion F. Cubitt, Anna Vossenkämper, and Peter M. Irving
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0301 basic medicine ,Adult ,Male ,medicine.medical_specialty ,medicine.medical_treatment ,lcsh:Medicine ,Inflammation ,Gastroenterology ,Antibodies ,Article ,03 medical and health sciences ,Ileostomy ,0302 clinical medicine ,Oral administration ,Internal medicine ,medicine ,Humans ,Colitis ,lcsh:Science ,Aged ,Lamina propria ,Multidisciplinary ,business.industry ,Tumor Necrosis Factor-alpha ,Stomach ,lcsh:R ,Middle Aged ,medicine.disease ,Ulcerative colitis ,Enteric coating ,Recombinant Proteins ,Intestines ,Crohn's disease ,030104 developmental biology ,medicine.anatomical_structure ,030211 gastroenterology & hepatology ,Colitis, Ulcerative ,Female ,lcsh:Q ,Immunotherapy ,medicine.symptom ,business ,medicine.drug - Abstract
V565 is an engineered TNFα-neutralising single domain antibody formulated into enteric coated mini-tablets to enable release in the intestine after oral administration as a possible oral treatment for inflammatory bowel disease (IBD). Following oral administration, ileal recovery of V565 was investigated in four patients with terminal ileostomy. Intestinal and systemic pharmacokinetics were measured in six patients with Crohn’s disease and evidence of target engagement assessed in five patients with ulcerative colitis. Following oral administration, V565 was detected at micromolar concentrations in ileal fluid from the ileostomy patients and in stools of the Crohn’s patients. In four of the five ulcerative colitis patients, biopsies taken after 7d dosing demonstrated V565 in the lamina propria with co-immunostaining on CD3+ T-lymphocytes and CD14+ macrophages. Phosphorylation of signalling proteins in biopsies taken after 7d oral dosing was decreased by approximately 50%. In conclusion, enteric coating of V565 mini-tablets provided protection in the stomach with gradual release in intestinal regions affected by IBD. Immunostaining revealed V565 tissue penetration and association with inflammatory cells, while decreased phosphoproteins after 7d oral dosing was consistent with V565-TNFα engagement and neutralising activity. Overall these results are encouraging for the clinical utility of V565 in the treatment of IBD.
- Published
- 2019
14. Discovery of a First-in-Class Receptor Interacting Protein 2 (RIP2) Kinase Specific Clinical Candidate, 2-((4-(Benzo[d]thiazol-5-ylamino)-6-(tert-butylsulfonyl)quinazolin-7-yl)oxy)ethyl Dihydrogen Phosphate, for the Treatment of Inflammatory Diseases
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Pamela A. Haile, Linda N. Casillas, Bartholomew J. Votta, Gren Z. Wang, Adam K. Charnley, Xiaoyang Dong, Michael J. Bury, Joseph J. Romano, John F. Mehlmann, Bryan W. King, Karl F. Erhard, Charles R. Hanning, David B. Lipshutz, Biva M. Desai, Carol A. Capriotti, Michelle C. Schaeffer, Scott B. Berger, Mukesh K. Mahajan, Michael A. Reilly, Rakesh Nagilla, Elizabeth J. Rivera, Helen H. Sun, John K. Kenna, Allison M. Beal, Michael T. Ouellette, Mike Kelly, Gillian Stemp, Máire A. Convery, Anna Vossenkämper, Thomas T. MacDonald, Peter J. Gough, John Bertin, and Robert W. Marquis
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Drug Discovery ,Molecular Medicine - Published
- 2019
15. The Food Additive Maltodextrin Promotes Endoplasmic Reticulum Stress–Driven Mucus Depletion and Exacerbates Intestinal InflammationSummary
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Massimo Federici, Francesca Ceccherini-Silberstein, Irene Marafini, Thomas T. MacDonald, Alfredo Colantoni, Antonio Di Grazia, Giovanni Monteleone, Vincenzo Dinallo, Maria Mavilio, Carmine Stolfi, Claudia Alteri, Federica Laudisi, Francesca Guerrieri, Davide Di Fusco, Angela Ortenzi, and Ivan Monteleone
- Subjects
0301 basic medicine ,Colitis ,IBD ,Intestinal Epithelium ,Unfolded Protein Response ,Settore MED/09 - Medicina Interna ,p38 mitogen-activated protein kinases ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,medicine ,Inositol ,lcsh:RC799-869 ,Settore MED/12 - Gastroenterologia ,Hepatology ,Settore BIO/12 ,Endoplasmic reticulum ,Gastroenterology ,Tauroursodeoxycholic acid ,medicine.disease ,Mucus ,Intestinal epithelium ,Cell biology ,030104 developmental biology ,chemistry ,Unfolded protein response ,030211 gastroenterology & hepatology ,lcsh:Diseases of the digestive system. Gastroenterology - Abstract
Background & Aims: Food additives, such as emulsifiers, stabilizers, or bulking agents, are present in the Western diet and their consumption is increasing. However, little is known about their potential effects on intestinal homeostasis. In this study we examined the effect of some of these food additives on gut inflammation. Methods: Mice were given drinking water containing maltodextrin (MDX), propylene glycol, or animal gelatin, and then challenged with dextran sulfate sodium or indomethacin. In parallel, mice fed a MDX-enriched diet were given the endoplasmic reticulum (ER) stress inhibitor tauroursodeoxycholic acid (TUDCA). Transcriptomic analysis, real-time polymerase chain reaction, mucin-2 expression, phosphorylated p38 mitogen-activated protein (MAP) kinase quantification, and H&E staining was performed on colonic tissues. Mucosa-associated microbiota composition was characterized by 16S ribosomal RNA sequencing. For the in vitro experiments, murine intestinal crypts and the human mucus-secreting HT29-methotrexate treated cell line were stimulated with MDX in the presence or absence of TUDCA or a p38 MAP kinase inhibitor. Results: Diets enriched in MDX, but not propylene glycol or animal gelatin, exacerbated intestinal inflammation in both models. Analysis of the mechanisms underlying the detrimental effect of MDX showed up-regulation of inositol requiring protein 1β, a sensor of ER stress, in goblet cells, and a reduction of mucin-2 expression with no significant change in mucosa-associated microbiota. Stimulation of murine intestinal crypts and HT29-methotrexate treated cell line cells with MDX induced inositol requiring protein 1β via a p38 MAP kinase–dependent mechanism. Treatment of mice with TUDCA prevented mucin-2 depletion and attenuated colitis in MDX-fed mice. Conclusions: MDX increases ER stress in gut epithelial cells with the downstream effect of reducing mucus production and enhancing colitis susceptibility. Keywords: Colitis, IBD, Unfolded Protein Response, Intestinal Epithelium
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- 2019
16. 795: AN INTERLEUKIN 22/CHEMOKINE AXIS DRIVES COLONIC NEUTROPHIL RECRUITMENT AND TREATMENT RESISTANCE IN ULCERATIVE COLITIS
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Polychronis Pavlidis, Anastasia Tsakmaki, Eirini Pantazi, Katherine Li, Domenico Cozzetto, Feifei Yang, Jonathan W. Lo, Ellie Alberts, Ana Caroline Costa Sa, Umar Niazi, Joshua Friedman, Anna Long, Yuchun Ding, Christopher Carey, Christopher A. Lamb, Mansoor Saqi, Matthew Madgwick, Lejla Gul, Agatha Treveil, Tamas Korcsmaros, Thomas T. Macdonald, Graham Lord, Gavin Bewick, and Nick Powell
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Hepatology ,Gastroenterology - Published
- 2022
17. T lymphocyte populations within the lamina propria
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Antonio Di Sabatino and Thomas T. MacDonald
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Pathology ,medicine.medical_specialty ,Lamina propria ,medicine.anatomical_structure ,medicine ,T lymphocyte ,Biology - Published
- 2020
18. Interleukin-22 orchestrates a pathological endoplasmic reticulum stress response transcriptional programme in colonic epithelial cells
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Behdad Afzali, Joshua R. Friedman, Domenico Cozzetto, Svetlana Saveljeva, Feifei Yang, Anastasia Tsakmaki, Rami Mohamed, Tsui Tjir-Li, Katherine Li, Emilie Stolarczyk, Thomas T. MacDonald, Aimee Parker, Polychronis Pavlidis, Carmen Pin, Gavin A. Bewick, Arthur Kaser, Eirini Pantazi, Danielle Minns, Graham M. Lord, Nick Powell, Jonathan Digby-Bell, Paul Lavender, Powell, Nick [0000-0003-3231-6950], Pavlidis, Polychronis [0000-0003-4864-2208], Friedman, Joshua [0000-0001-9382-8429], Bewick, Gavin A [0000-0002-4335-8403], Apollo - University of Cambridge Repository, and Wellcome Trust
- Subjects
0301 basic medicine ,Transcription, Genetic ,Apoptosis ,THERAPY ,Inflammatory bowel disease ,Interleukin-23 ,DISEASE ,Interleukin 22 ,Mice ,0302 clinical medicine ,Crohn Disease ,Medicine ,Intestinal Mucosa ,INDUCED APOPTOSIS ,Gastrointestinal agent ,INDUCTION ,Tunicamycin ,Interleukin-17 ,Gastroenterology ,Colitis ,Endoplasmic Reticulum Stress ,Phenylbutyrates ,Recombinant Proteins ,3. Good health ,Anti-Bacterial Agents ,Organoids ,030220 oncology & carcinogenesis ,Ustekinumab ,medicine.symptom ,ER stress ,Life Sciences & Biomedicine ,INTESTINAL INFLAMMATION ,Cell Survival ,Colon ,INNATE LYMPHOID-CELLS ,Inflammation ,Phenylbutyrate ,MECHANISMS ,03 medical and health sciences ,Gastrointestinal Agents ,inflammatory bowel disease ,Animals ,Humans ,Science & Technology ,Gastroenterology & Hepatology ,business.industry ,Endoplasmic reticulum ,Interleukins ,Recent Advances in Basic Science ,Patient Acuity ,1103 Clinical Sciences ,Epithelial Cells ,medicine.disease ,digestive system diseases ,Disease Models, Animal ,030104 developmental biology ,PROTECT ,Chronic Disease ,Unfolded protein response ,Cancer research ,Unfolded Protein Response ,1114 Paediatrics and Reproductive Medicine ,business - Abstract
ObjectiveThe functional role of interleukin-22 (IL22) in chronic inflammation is controversial, and mechanistic insights into how it regulates target tissue are lacking. In this study, we evaluated the functional role of IL22 in chronic colitis and probed mechanisms of IL22-mediated regulation of colonic epithelial cells.DesignTo investigate the functional role of IL22 in chronic colitis and how it regulates colonic epithelial cells, we employed a three-dimentional mini-gut epithelial organoid system, in vivo disease models and transcriptomic datasets in human IBD.ResultsAs well as inducing transcriptional modules implicated in antimicrobial responses, IL22 also coordinated an endoplasmic reticulum (ER) stress response transcriptional programme in colonic epithelial cells. In the colon of patients with active colonic Crohn’s disease (CD), there was enrichment of IL22-responsive transcriptional modules and ER stress response modules. Strikingly, in an IL22-dependent model of chronic colitis, targeting IL22 alleviated colonic epithelial ER stress and attenuated colitis. Pharmacological modulation of the ER stress response similarly impacted the severity of colitis. In patients with colonic CD, antibody blockade of IL12p40, which simultaneously blocks IL12 and IL23, the key upstream regulator of IL22 production, alleviated the colonic epithelial ER stress response.ConclusionsOur data challenge perceptions of IL22 as a predominantly beneficial cytokine in IBD and provide novel insights into the molecular mechanisms of IL22-mediated pathogenicity in chronic colitis. Targeting IL22-regulated pathways and alleviating colonic epithelial ER stress may represent promising therapeutic strategies in patients with colitis.Trial registration numberNCT02749630.
- Published
- 2020
19. A functional IL1RL1 variant regulates corticosteroid-induced sST2 expression in ulcerative colitis
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Karen Dubois-Camacho, Jonás Chnaiderman, Lucía Núñez, Thomas T. MacDonald, Alejandro Torres-Riquelme, Daniela Simian, María-Julieta González, Marjorie De la Fuente, Martin Montecino, John A. Cidlowski, Paulina A García-González, Rodrigo Quera, Anna Vossenkämper, Hugo Sepulveda, Marcela A. Hermoso, and David Díaz-Jiménez
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0301 basic medicine ,Male ,IL1RL1 ,lcsh:Medicine ,Dexamethasone ,Intestinal mucosa ,Transcription (biology) ,Adrenal Cortex Hormones ,Intestinal Mucosa ,Receptor ,Promoter Regions, Genetic ,lcsh:Science ,IN-VIVO ,Cells, Cultured ,Multidisciplinary ,Middle Aged ,CROHNS-DISEASE ,Up-Regulation ,MAST-CELLS ,Female ,medicine.drug ,Adult ,SUSCEPTIBILITY LOCI ,DNA-BINDING ,Biology ,Polymorphism, Single Nucleotide ,Article ,03 medical and health sciences ,Downregulation and upregulation ,medicine ,Humans ,Genetic Predisposition to Disease ,Colitis ,SOLUBLE ST2 ,lcsh:R ,HUMAN GLUCOCORTICOID RECEPTOR ,Promoter ,Sequence Analysis, DNA ,medicine.disease ,Molecular biology ,Interleukin-1 Receptor-Like 1 Protein ,PROMOTER USAGE ,030104 developmental biology ,Gene Expression Regulation ,BARRIER FUNCTION ,Cancer research ,Colitis, Ulcerative ,lcsh:Q ,INFLAMMATORY-BOWEL-DISEASE - Abstract
The ST2/IL33 signalling pathway has been associated with ulcerative colitis (UC). ST2, encoded by the IL1RL1 gene, is expressed as both a membrane-anchored receptor (ST2L) activated by IL33 and as a soluble receptor (sST2) with anti-inflammatory properties. In UC patients, sST2 is further increased by corticosteroid treatment; however, the glucocorticoid-mediated molecular regulation remains unknown. We therefore tested whether genetic variants in the IL1RL1 distal promoter are involved in UC and affect glucocorticoid-mediated ST2 expression. Serum ST2 levels and genetic variants in the IL1RL1 distal promoter were examined by ELISA and PCR sequencing in UC patients receiving corticosteroids. Glucocorticoid-mediated ST2 production was evaluated in intestinal mucosa cultures. Molecular regulation of glucocorticoid-mediated ST2 was assessed by RT-qPCR, ChIP assay and luciferase reporter assay. Dexamethasone effect on ST2 transcript expression was analyzed in leukocytes and related to IL1RL1 variants. Sequencing of a distal IL1RL1 promoter region demonstrated that SNPs rs6543115(C) and rs6543116(A) are associated with increased sST2 in UC patients on corticosteroids. Dexamethasone up-regulated sST2 transcription through interaction with the glucocorticoid-response element (GRE) carrying rs6543115(C) variant. Our data indicate that IL1RL1 SNPs rs6543115(C) confer susceptibility to UC and is contained in the GRE, which may modulate glucocorticoid-induced sST2 expression.
- Published
- 2017
20. Homeostasis of the gut barrier and potential biomarkers
- Author
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Agnes Meheust, Muriel Derrien, Patrice D. Cani, Freddy J. Troost, Thomas T. MacDonald, Vassilia Theodorou, Jerry M. Wells, Robert-Jan M. Brummer, Annick Mercenier, Willem M. de Vos, Jan Dekker, Clara Lucia Garcia-Rodenas, Arjen Nauta, Animal Sciences, Host-Microbe Interactomics, Wageningen University and Research Center (WUR), School of Medicine and Health, Nutrition-Gut-Brain Interactions Research Centre, Örebro University, Centre Daniel Carasso, Danone Nutricia Research, Barts and The London School of Medicine and Dentistry, Blizard Institute, Queen Mary University of London (QMUL), University Hospital Maastricht, Department of Internal Medicine, Division of Gastroenterology-Hepatology, Maastricht University Medical Center (MUMC), Maastricht University [Maastricht]-Maastricht University [Maastricht], Metabolism and Nutrition Research Group, Walloon Excellence in Life Sciences and BIOtechnology (WELBIO), Louvain Drug Research Institute, Université Catholique de Louvain, ToxAlim (ToxAlim), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Ecole Nationale Vétérinaire de Toulouse (ENVT), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Ecole d'Ingénieurs de Purpan (INPT - EI Purpan), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Recherche Agronomique (INRA), Groupe Danone, Laboratory of Microbiology, CHU Rouen, Normandie Université (NU)-Normandie Université (NU), Institute of Nutritional Science, Nestlé Research Center, FrieslandCampina, Wageningen University and Research [Wageningen] (WUR), Maastricht University Medical Centre (MUMC), Université Catholique de Louvain = Catholic University of Louvain (UCL), Neuro-Gastroentérologie & Nutrition (ToxAlim-NGN), Institut National de la Recherche Agronomique (INRA)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Ecole Nationale Vétérinaire de Toulouse (ENVT), Université Fédérale Toulouse Midi-Pyrénées-Ecole d'Ingénieurs de Purpan (INPT - EI Purpan), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Recherche Agronomique (INRA)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées, UCL - SSS/LDRI - Louvain Drug Research Institute, and Wells, Jerry M.
- Subjects
0301 basic medicine ,IRRITABLE-BOWEL-SYNDROME ,Gastrointestinal Diseases ,Physiology ,epithelial permeability ,Segmented filamentous bacteria ,[SDV]Life Sciences [q-bio] ,BACTERICIDAL/PERMEABILITY-INCREASING PROTEIN ,toxicologie alimentaire ,microbiote digestif ,POLYMERIC IMMUNOGLOBULIN RECEPTOR ,Review ,human health ,peptide antimicrobien ,antimicrobial peptides ,Epithelial permeability ,Microbiologie ,Gut barrier ,TIGHT JUNCTION PERMEABILITY ,Homeostasis ,IGA MONOCLONAL-ANTIBODIES ,biology ,Microbiota ,Gastroenterology ,Pattern recognition receptor ,santé humaine ,3. Good health ,INNATE IMMUNE-RESPONSE ,trouble gastrointestinal ,perméabilité intestinale ,Antimicrobial peptides ,moyen de prévention ,Microbiology ,03 medical and health sciences ,Immune system ,Physiology (medical) ,medicine ,microbiota ,Animals ,Humans ,Host-Microbe Interactomics ,REGULATORY T-CELLS ,gut barrier ,VLAG ,TOLL-LIKE RECEPTORS ,Innate immune system ,Hepatology ,barrière intestinale ,medicine.disease ,Bactericidal/permeability-increasing protein ,Mucus ,Gastrointestinal Tract ,SEGMENTED FILAMENTOUS BACTERIA ,030104 developmental biology ,Immunology ,biology.protein ,WIAS ,INTESTINAL EPITHELIAL-CELLS ,Dysbiosis - Abstract
The gut barrier plays a crucial role by spatially compartmentalizing bacteria to the lumen through the production of secreted mucus and is fortified by the production of secretory IgA (sIgA) and antimicrobial peptides and proteins. With the exception of sIgA, expression of these protective barrier factors is largely controlled by innate immune recognition of microbial molecular ligands. Several specialized adaptations and checkpoints are operating in the mucosa to scale the immune response according to the threat and prevent overreaction to the trillions of symbionts inhabiting the human intestine. A healthy microbiota plays a key role influencing epithelial barrier functions through the production of short-chain fatty acids (SCFAs) and interactions with innate pattern recognition receptors in the mucosa, driving the steady-state expression of mucus and antimicrobial factors. However, perturbation of gut barrier homeostasis can lead to increased inflammatory signaling, increased epithelial permeability, and dysbiosis of the microbiota, which are recognized to play a role in the pathophysiology of a variety of gastrointestinal disorders. Additionally, gut-brain signaling may be affected by prolonged mucosal immune activation, leading to increased afferent sensory signaling and abdominal symptoms. In turn, neuronal mechanisms can affect the intestinal barrier partly by activation of the hypothalamus-pituitary-adrenal axis and both mast cell-dependent and mast cell-independent mechanisms. The modulation of gut barrier function through nutritional interventions, including strategies to manipulate the microbiota, is considered a relevant target for novel therapeutic and preventive treatments against a range of diseases. Several biomarkers have been used to measure gut permeability and loss of barrier integrity in intestinal diseases, but there remains a need to explore their use in assessing the effect of nutritional factors on gut barrier function. Future studies should aim to establish normal ranges of available biomarkers and their predictive value for gut health in human cohorts.
- Published
- 2017
21. Discovery of a First-in-Class Receptor Interacting Protein 1 (RIP1) Kinase Specific Clinical Candidate (GSK2982772) for the Treatment of Inflammatory Diseases
- Author
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Xiaoyang Dong, Anna Vossenkämper, Attiq Rahman, Viera Kasparcova, Robert Singhaus, Joshua N. Finger, Bryan W. King, Patrick M. Eidam, Michael Reilly, David D. Wisnoski, Scott B. Berger, John D. Lich, James Kang, Michael T. Ouellette, Lauren Dare, Philip A. Harris, Julie A. Cox, Nathan A. Miller, John Bertin, Michelle C. Schaeffer, Deepak Bandyopadhyay, Daohua Zhang, Alan R. Rendina, Clark A. Sehon, Sandra J. Hoffman, Peter J. Gough, Rachel D. Totoritis, Thomas T. MacDonald, Yunfeng Lan, Paris Ward, Barbara A. Swift, Robert W. Marquis, Lara Kathryn Leister, Rakesh Nagilla, Elizabeth J. Rivera, Helen H. Sun, Carol A. Capriotti, Nino Campobasso, Ruth Lehr, Christina S. Pao, and Jae U. Jeong
- Subjects
0301 basic medicine ,Swine ,Necroptosis ,Anti-Inflammatory Agents ,Inflammation ,Pharmacology ,Mice ,03 medical and health sciences ,Dogs ,0302 clinical medicine ,Psoriasis ,Drug Discovery ,medicine ,Animals ,Humans ,Potency ,Protein Kinase Inhibitors ,Tumor Necrosis Factor-alpha ,Chemistry ,Kinase ,Haplorhini ,Benzazepines ,medicine.disease ,Ulcerative colitis ,Rats ,Molecular Docking Simulation ,030104 developmental biology ,Receptor-Interacting Protein Serine-Threonine Kinases ,030220 oncology & carcinogenesis ,Rheumatoid arthritis ,Cytokines ,Swine, Miniature ,Molecular Medicine ,Colitis, Ulcerative ,Rabbits ,medicine.symptom ,RECEPTOR-INTERACTING PROTEIN - Abstract
RIP1 regulates necroptosis and inflammation and may play an important role in contributing to a variety of human pathologies, including immune-mediated inflammatory diseases. Small-molecule inhibitors of RIP1 kinase that are suitable for advancement into the clinic have yet to be described. Herein, we report our lead optimization of a benzoxazepinone hit from a DNA-encoded library and the discovery and profile of clinical candidate GSK2982772 (compound 5), currently in phase 2a clinical studies for psoriasis, rheumatoid arthritis, and ulcerative colitis. Compound 5 potently binds to RIP1 with exquisite kinase specificity and has excellent activity in blocking many TNF-dependent cellular responses. Highlighting its potential as a novel anti-inflammatory agent, the inhibitor was also able to reduce spontaneous production of cytokines from human ulcerative colitis explants. The highly favorable physicochemical and ADMET properties of 5, combined with high potency, led to a predicted low oral dose in humans.
- Published
- 2017
22. Regulation of human intestinal T-cell responses by type 1 interferon-STAT1 signaling is disrupted in inflammatory bowel disease
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J. Lung, M. Pathak, Andrew J. Stagg, Thomas T. MacDonald, Edward M. Giles, Theodore J. Sanders, James O. Lindsay, and Neil E. McCarthy
- Subjects
Male ,0301 basic medicine ,Adolescent ,Colon ,medicine.medical_treatment ,Immunology ,Inflammation ,Lymphocyte Activation ,T-Lymphocytes, Regulatory ,Inflammatory bowel disease ,Proinflammatory cytokine ,Immunomodulation ,Interferon-gamma ,Mice ,03 medical and health sciences ,Interferon ,medicine ,Animals ,Humans ,Immunology and Allergy ,Interferon gamma ,STAT1 ,Phosphorylation ,Antibodies, Blocking ,Child ,Cells, Cultured ,biology ,business.industry ,Cell Differentiation ,Interferon-beta ,Inflammatory Bowel Diseases ,medicine.disease ,Interleukin-10 ,Interleukin 10 ,STAT1 Transcription Factor ,030104 developmental biology ,Cytokine ,biology.protein ,Female ,medicine.symptom ,business ,Signal Transduction ,medicine.drug - Abstract
Type 1 interferon (IFN-1) promotes regulatory T-cell function to suppress inflammation in the mouse intestine, but little is known about IFN-1 in the human gut. We therefore assessed the influence of IFN-1 on CD4+ T-cells isolated from human colon tissue obtained from healthy controls or patients with inflammatory bowel disease (IBD). Immunofluorescent imaging revealed constitutive expression of IFNβ in human intestinal tissue, and colonic T-cells were responsive to exogenous IFN-1 as assessed by phosphorylation of signal transduction and activator of transcription 1 (pSTAT1) and induction of interferon stimulated genes (ISGs). Unlike their blood counterparts, intestinal T-cells from non-inflamed regions of IBD colon displayed enhanced responsiveness to IFN-1, increased frequency of pSTAT1+ cells, and greater induction of ISGs upon IFN-1 exposure in vitro. In healthy tissue, antibody neutralization of IFNβ selectively reduced T-cell production of the pro-regulatory cytokine interleukin-10 (IL-10) and increased IFNγ synthesis. In contrast, neutralization of IFNβ in IBD tissue cultures increased the frequency of T-cells producing inflammatory cytokines but did not alter IL-10 expression. These data support a role for endogenous IFN-1 as a context-dependent modulator of T-cell function that promotes regulatory activity in healthy human intestine, but indicate that the IFN-1/STAT1 pathway is dysregulated in inflammatory bowel disease.
- Published
- 2017
23. Host defences to Citrobacter rodentium
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Thomas T. MacDonald, Nathalie S. Gonçalves, Gad Frankel, Cameron P. Simmons, and Gordon Dougan
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Microbiology (medical) ,Colon ,Inflammation ,Biology ,medicine.disease_cause ,Microbiology ,Mice ,Immunity ,medicine ,Citrobacter rodentium ,Animals ,Humans ,Secretion ,Adhesins, Bacterial ,Escherichia coli ,Pathogen ,Immunity, Mucosal ,Intimin ,Hyperplasia ,Escherichia coli Proteins ,Enterobacteriaceae Infections ,General Medicine ,Bacterial adhesin ,Citrobacter freundii ,Mice, Inbred C57BL ,Disease Models, Animal ,Infectious Diseases ,Immunology ,medicine.symptom ,Carrier Proteins - Abstract
Citrobacter rodentium is a natural non-invasive bacterial pathogen which infects the distal colon of mice. It uses the same molecular mechanisms of type III secretion as human enteropathogenic and enterohemorrhagic Escherichia coli to colonise the epithelial cells of the gut and is therefore an ideal model to study host-bacterial pathogen interactions in vivo. Infection elicits mucosal inflammation with similarities to inflammatory bowel disease, and so it is a readily accessible model to investigate the relationship between inflammation and anti-bacterial immunity in the gut.
- Published
- 2019
24. SAT-LB056 Is AIP a Tumor Suppressor or an Oncogene? AIP as a Novel Regulator of the Oncogene BCL6 in Diffuse Large B Cell Lymphoma
- Author
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Dijue Sun, Christopher D. Buckley, Xinyu Wan, Urszula Stopka-Farooqui, Oliver Haworth, Melania Capasso, Anna Vossenkämper, Sayka Barry, Márta Korbonits, Sherine Norris, Andrew Clear, Gregory Parsonage, John G. Gribben, Ezra Aksoy, Thomas T. MacDonald, and Jennifer L. Marshall
- Subjects
Oncogene ,Endocrinology, Diabetes and Metabolism ,Regulator ,Tumor Biology of Breast and Prostate Cancers ,Biology ,BCL6 ,medicine.disease ,law.invention ,immune system diseases ,law ,hemic and lymphatic diseases ,medicine ,Cancer research ,Tumor Biology ,Suppressor ,Diffuse large B-cell lymphoma - Abstract
Background: Heterozygous germline loss-of-function mutations in the aryl hydrocarbon receptor interacting protein (AIP) gene are usually associated with acromegaly or gigantism due to a young-onset somatotropinoma. AIP functions as a tumor suppressor gene in the pituitary, while its role in other tumours is unknown. AIP is highly expressed in patients with Diffuse Large B Cell Lymphoma (DLBCL) compared to any other tumor type. DLBCL arises from germinal centre B cells which characteristically express the transcriptional repressor B cell lymphoma-6 (BCL6), key for germinal centre formation. Increased expression of BCL6 can lead to the development of DLBCL. Despite the obvious importance of BCL6 expression in GC B cells, the mechanisms by which it is regulated are still poorly understood. Aim: The aim of this study was to investigate the relationship of the high AIP expression, BCL6 and the pathobiology of DLBCL. Methods: We generated mice carrying a conditional homozygous deletion of Aip in T and B cells (Aipfl/fl;Rag1Cre/+). Co-localization of AIP and members of the BCL6 ubiquitination/proteasome pathway by immunofluorescence analysis. Ubiquitin-mediated proteasomal degradation study was done by immuno-precipitation (IP). Results: Our study found that AIP is highly expressed in DLBCL. Genetic deletion of Aip revealed that AIP supported BCL6 expression in mice. The ubiquitin E3 ligase FBXO11 has been previously shown to add ubiquitin to BCL6. We found that AIP could bind to the deubiquitinase UCHL1 and that UCHL1 could remove ubiquitin from BCL6 thereby supporting BCL6 expression by preventing FBXO11 mediated degradation of BCL6 recapitulating what we observed in Aip deficient B cells. Conclusions: The data presented here reveal AIP to be a novel positive regulator of BCL6 protein expression which is commonly up-regulated in DLBCL and prevents FBXO11 degradation of BCL6 by enabling the UCHL1 to remove ubiquitin from BCL6. Therefore, AIP is a potential novel therapeutic target to treat DLBCL. It appears that AIP function as a tumor suppressor in the pituitary and as an oncogene to the pathobiology of DLBCL. Unless otherwise noted, all abstracts presented at ENDO are embargoed until the date and time of presentation. For oral presentations, the abstracts are embargoed until the session begins. Abstracts presented at a news conference are embargoed until the date and time of the news conference. The Endocrine Society reserves the right to lift the embargo on specific abstracts that are selected for promotion prior to or during ENDO.
- Published
- 2019
25. Preclinical evaluation of EPHX2 inhibition as a novel treatment for inflammatory bowel disease
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Vinod Kumar, Qing Xie, Anna Vossenkämper, Bao Hoang, Mark E. Burgert, Mike Reilly, Lin Wang, Wensheng Xie, Mark R. Hurle, Bart Votta, Neetu Rajpal, Pankaj K. Agarwal, Xin Zeng, Thomas T. MacDonald, Deepak K. Rajpal, Sarah O’Donnell, William C. Reisdorf, and Yolanda Sanchez
- Subjects
0301 basic medicine ,Epoxide hydrolase 2 ,Science ,Drug Evaluation, Preclinical ,Inflammation ,Pharmacology ,Inflammatory bowel disease ,Proinflammatory cytokine ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Immune system ,medicine ,Animals ,Humans ,Epoxide Hydrolases ,Cyclohexylamines ,Multidisciplinary ,Triazines ,business.industry ,Dextran Sulfate ,Colitis ,Inflammatory Bowel Diseases ,medicine.disease ,Ulcerative colitis ,Mice, Inbred C57BL ,Disease Models, Animal ,Interleukin 10 ,030104 developmental biology ,030220 oncology & carcinogenesis ,Cytokines ,Medicine ,Female ,Tumor necrosis factor alpha ,medicine.symptom ,business - Abstract
Epoxyeicosatrienoic acids (EETs) are signaling lipids produced by cytochrome P450 epoxygenation of arachidonic acid, which are metabolized by EPHX2 (epoxide hydrolase 2, alias soluble epoxide hydrolase or sEH). EETs have pleiotropic effects, including anti-inflammatory activity. Using a Connectivity Map (CMAP) approach, we identified an inverse-correlation between an exemplar EPHX2 inhibitor (EPHX2i) compound response and an inflammatory bowel disease patient-derived signature. To validate the gene-disease link, we tested a pre-clinical tool EPHX2i (GSK1910364) in a mouse disease model, where it showed improved outcomes comparable to or better than the positive control Cyclosporin A. Up-regulation of cytoprotective genes and down-regulation of proinflammatory cytokine production were observed in colon samples obtained from EPHX2i-treated mice. Follow-up immunohistochemistry analysis verified the presence of EPHX2 protein in infiltrated immune cells from Crohn's patient tissue biopsies. We further demonstrated that GSK2256294, a clinical EPHX2i, reduced the production of IL2, IL12p70, IL10 and TNFα in both ulcerative colitis and Crohn's disease patient-derived explant cultures. Interestingly, GSK2256294 reduced IL4 and IFNγ in ulcerative colitis, and IL1β in Crohn's disease specifically, suggesting potential differential effects of GSK2256294 in these two diseases. Taken together, these findings suggest a novel therapeutic use of EPHX2 inhibition for IBD.
- Published
- 2019
26. The IL-33/ST2 axis: Role in health and disease
- Author
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Thomas T. MacDonald, Marjorie De la Fuente, and Marcela A. Hermoso
- Subjects
Cell type ,Endocrinology, Diabetes and Metabolism ,Immunology ,Interleukin-1 Receptor-Like 1 Protein ,IL1RL1 ,Autoimmunity ,Receptors, Cell Surface ,Inflammation ,medicine.disease_cause ,Skin Diseases ,General Biochemistry, Genetics and Molecular Biology ,Pathogenesis ,Neoplasms ,medicine ,Animals ,Humans ,Immunology and Allergy ,Receptor ,business.industry ,Inflammatory Bowel Diseases ,Interleukin-33 ,Cell biology ,medicine.symptom ,Signal transduction ,business ,Signal Transduction - Abstract
IL-33, an IL-1 family member, is expressed by many cell types and can regulate gene transcription. IL-33 is released upon cell necrosis and the precursor form is enzymatically processed, and then drives inflammation as a damage-associated molecular pattern. The IL-33 receptor ST2, encoded by IL1RL1, is expressed as both a membrane-anchored receptor (ST2L) activated by IL-33, and as a soluble variant (sST2) that exhibits anti-inflammatory properties. The IL-33/ST2 axis is involved in the pathogenesis of atopic and autoimmune diseases, cancer, and central nervous system disorders. Here, we review recent findings on the role of the IL-33/ST2 axis in health and disease.
- Published
- 2015
27. ADTU-02 V565, a novel oral anti-TNF domain antibody, reduces colonic mucosal inflammation in patients with UC
- Author
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Thomas T. MacDonald, Suhail Nurbhai, Michael R. West, Anna Vossenkaemper, Gareth Parkes, Kausik K. Ray, and Scott Crowe
- Subjects
medicine.medical_specialty ,Lamina propria ,medicine.diagnostic_test ,business.industry ,02 engineering and technology ,021001 nanoscience & nanotechnology ,medicine.disease ,030226 pharmacology & pharmacy ,Gastroenterology ,Ulcerative colitis ,Inflammatory bowel disease ,Infliximab ,Proinflammatory cytokine ,03 medical and health sciences ,0302 clinical medicine ,medicine.anatomical_structure ,Mucosal immunology ,Internal medicine ,Biopsy ,Medicine ,Tumor necrosis factor alpha ,0210 nano-technology ,business ,medicine.drug - Abstract
Introduction Monoclonal antibodies to TNF transformed treatment options for patients with Inflammatory Bowel Disease (IBD). V565 is a novel oral anti-TNF domain antibody (Vorabody) engineered to be resistant to intestinal proteases. It is in development as a potential oral treatment for IBD. In vitro it suppressed phosphorylation of tyrosine kinases and signalling proteins and inhibited the release of inflammatory cytokines following culture with biopsies taken from patients with CD (Crowe et al. 18th International Congress of Mucosal Immunology, July 19–22 2017, Washington DC, USA). It was safe and well tolerated after high single and multiple doses in healthy volunteers and patient volunteers with CD and resulted in high concentrations of active drug in ileal fluid and faeces. Aims & Methods This open label study was designed to demonstrate that V565 enters GI mucosa and exerts a beneficial effect on inflammatory processes following oral dosing for 7 days to patients with Ulcerative Colitis. Patients with a Mayo score of 3–10 including an endoscopy score of ≥1 had up to 7 days of oral dosing with 555 mg tid V565. Sigmoidoscopy with biopsies was performed before and after the dosing period. The primary outcomes of interest were presence of V565 in the mucosa and reduction from baseline in phosphorylation of tyrosine kinases and signalling proteins. Detection of V565 was determined by immunohistochemistry. Phosphorylation was determined using PathScan RTK signalling arrays (Vossenkaemper et al 2014. Gastroenterology 147:172–83). Results Five patient volunteers were treated Due to visit scheduling, most received 6 days treatment. Presence of V565 was confirmed in the inflamed lamina propria and co-localised with CD14 +macrophages in post-treatment biopsies. Overall phosphorylation of the panel of kinases and signalling proteins was reduced by approximately 50% in four of the five patients. There were no treatment induced ADAs. Conclusion V565, an oral anti-TNF domain antibody engineered to be resistant to intestinal proteases, was demonstrated bound to CD14 +macrophages in the lamina propria of UC patients and resulted in inhibition of mucosal inflammatory processes after 6–7 days oral dosing. The reduction of 50% in overall phosphorylation is similar to that seen in an earlier study of UC biopsy cultures with infliximab at a concentration of 67 nM (10 µg/ml), a serum concentration associated with mucosal healing (Ungar et al, Clin Gastroenterol Hepatol. 2016 Apr;14(4):550–557). These results provide encouragement that oral dosing with V565 will be a beneficial oral treatment option for patients with IBD.
- Published
- 2018
28. Aryl Hydrocarbon Receptor Interacting Protein Maintains Germinal Center B Cells through Suppression of BCL6 Degradation
- Author
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John G. Gribben, Jennifer L. Marshall, Sherine Norris, Urszula Stopka-Farooqui, Anna Vossenkämper, Thomas T. MacDonald, Dijue Sun, Gregory Parsonage, Andrew Clear, Melania Capasso, Christopher D. Buckley, Xinyu Wan, Sayka Barry, Oliver Haworth, Márta Korbonits, and Ezra Aksoy
- Subjects
0301 basic medicine ,Adult ,Male ,Proteasome Endopeptidase Complex ,Protein-Arginine N-Methyltransferases ,B-cell receptor ,General Biochemistry, Genetics and Molecular Biology ,Article ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Immune system ,immune system diseases ,Heat shock protein ,hemic and lymphatic diseases ,medicine ,Animals ,Humans ,ddc:610 ,lcsh:QH301-705.5 ,Protein kinase B ,B cell ,Aged ,Aged, 80 and over ,B-Lymphocytes ,Chemistry ,F-Box Proteins ,Intracellular Signaling Peptides and Proteins ,Ubiquitination ,Germinal center ,Middle Aged ,BCL6 ,medicine.disease ,Germinal Center ,Cell biology ,030104 developmental biology ,medicine.anatomical_structure ,HEK293 Cells ,lcsh:Biology (General) ,Proteolysis ,Proto-Oncogene Proteins c-bcl-6 ,Female ,Lymphoma, Large B-Cell, Diffuse ,Diffuse large B-cell lymphoma ,Ubiquitin Thiolesterase ,030217 neurology & neurosurgery ,Protein Binding - Abstract
Summary: B cell lymphoma-6 (BCL6) is highly expressed in germinal center B cells, but how its expression is maintained is still not completely clear. Aryl hydrocarbon receptor interacting protein (AIP) is a co-chaperone of heat shock protein 90. Deletion of Aip in B cells decreased BCL6 expression, reducing germinal center B cells and diminishing adaptive immune responses. AIP was required for optimal AKT signaling in response to B cell receptor stimulation, and AIP protected BCL6 from ubiquitin-mediated proteasomal degradation by the E3-ubiquitin ligase FBXO11 by binding to the deubiquitinase UCHL1, thus helping to maintain the expression of BCL6. AIP was highly expressed in primary diffuse large B cell lymphomas compared to healthy tissue and other tumors. Our findings describe AIP as a positive regulator of BCL6 expression with implications for the pathobiology of diffuse large B cell lymphoma. : BCL6 overexpression contributes to the pathobiology of diffuse large B cell lymphoma (DLBCL). Sun et al. find that the co-chaperone aryl hydrocarbon receptor interacting protein (AIP), whose high expression is associated with reduced survival of DLBCL patients, helps maintain BCL6 expression by facilitating the removal of ubiquitin from BCL6. Keywords: AIP, BCL6, FBXO11, UCHL1, ubiquitination, lymphoma
- Published
- 2018
29. Preclinical Development of a Novel, Orally-Administered Anti-Tumour Necrosis Factor Domain Antibody for the Treatment of Inflammatory Bowel Disease
- Author
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Marion F. Cubitt, Michael R. West, Simon Clare, Luana Maggiore, Thomas T. MacDonald, Keith P. Ray, Timothy M. Carlton, Kevin J. Roberts, Katherine Harcourt, J. Scott Crowe, Anna Vossenkämper, and Jill Reckless
- Subjects
0301 basic medicine ,Male ,Colon ,medicine.medical_treatment ,Drug Evaluation, Preclinical ,lcsh:Medicine ,Administration, Oral ,Pharmacology ,Inflammatory bowel disease ,Article ,03 medical and health sciences ,Mice ,Oral administration ,Ileum ,Medicine ,Animals ,Humans ,Colitis ,Intestinal Mucosa ,lcsh:Science ,Multidisciplinary ,biology ,business.industry ,Tumor Necrosis Factor-alpha ,lcsh:R ,medicine.disease ,Inflammatory Bowel Diseases ,Infliximab ,3. Good health ,Disease Models, Animal ,030104 developmental biology ,Cytokine ,biology.protein ,Cytokines ,lcsh:Q ,Tumor necrosis factor alpha ,Antibody ,business ,Immunostaining ,Ex vivo ,Biomarkers - Abstract
TNFα is an important cytokine in inflammatory bowel disease. V565 is a novel anti-TNFα domain antibody developed for oral administration in IBD patients, derived from a llama domain antibody and engineered to enhance intestinal protease resistance. V565 activity was evaluated in TNFα-TNFα receptor-binding ELISAs as well as TNFα responsive cellular assays and demonstrated neutralisation of both soluble and membrane TNFα with potencies similar to those of adalimumab. Although sensitive to pepsin, V565 retained activity after lengthy incubations with trypsin, chymotrypsin, and pancreatin, as well as mouse small intestinal and human ileal and faecal supernatants. In orally dosed naïve and DSS colitis mice, high V565 concentrations were observed in intestinal contents and faeces and immunostaining revealed V565 localisation in mouse colon tissue. V565 was detected by ELISA in post-dose serum of colitis mice, but not naïve mice, demonstrating penetration of disrupted epithelium. In an ex vivo human IBD tissue culture model, V565 inhibition of tissue phosphoprotein levels and production of inflammatory cytokine biomarkers was similar to infliximab, demonstrating efficacy when present at the disease site. Taken together, results of these studies provide confidence that oral V565 dosing will be therapeutic in IBD patients where the mucosal epithelial barrier is compromised.
- Published
- 2018
30. Abnormal thymic stromal lymphopoietin expression in the duodenal mucosa of patients with coeliac disease
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Katerina Tsilingiri, Maria Rescigno, Alastair Forbes, A. Pasini, Paolo Giuffrida, Cinzia Papadia, Alessandro Vanoli, Giulia Fornasa, Sylvia L.F. Pender, Eleanor Wood, C. Ubezio, Paolo Biancheri, Thomas T. MacDonald, Antonio Di Sabatino, and Gino Roberto Corazza
- Subjects
Adult ,Male ,MATRIX METALLOPROTEINASE ,0301 basic medicine ,Thymic stromal lymphopoietin ,Duodenum ,Biopsy ,T-Lymphocytes ,GUT IMMUNOLOGY ,medicine.medical_treatment ,Statistics as Topic ,SMALL INTESTINE ,Fluorescent Antibody Technique ,Coeliac Disease ,Biology ,Immune tolerance ,03 medical and health sciences ,Thymic Stromal Lymphopoietin ,Intestinal mucosa ,Immune Tolerance ,medicine ,Humans ,Protein Isoforms ,Enteropathy ,Interleukin 8 ,Intestinal Mucosa ,Aged ,Furin ,Receptors, Interleukin-7 ,Interleukin-8 ,Gastroenterology ,Interleukin ,Middle Aged ,medicine.disease ,3. Good health ,Celiac Disease ,030104 developmental biology ,Cytokine ,MUCOSAL IMMUNITY ,Immunology ,Cytokines ,Female ,Ex vivo - Abstract
Objective The short isoform of thymic stromal lymphopoietin (TSLP), a cytokine constitutively expressed by epithelial cells, is crucial in preserving immune tolerance in the gut. TSLP deficiency has been implicated in sustaining intestinal damage in Crohn9s disease. We explored mucosal TSLP expression and function in refractory and uncomplicated coeliac disease (CD), a T-cell-mediated enteropathy induced by gluten in genetically susceptible individuals. Design TSLP isoforms—long and short—and receptors—TSLPR and interleukin (IL)-7Rα—were assessed by immunofluorescence, immunoblotting and qRT-PCR in the duodenum of untreated, treated, potential and refractory patients with CD. The ability of the serine protease furin or CD biopsy supernatants to cleave TSLP was evaluated by immunoblotting. The production of interferon (IFN)-γ and IL-8 by untreated CD biopsies cultured ex vivo with TSLP isoforms was also assessed. Results Mucosal TSLP, but not TSLPR and IL-7Rα, was reduced in untreated CD and refractory CD in comparison to treated CD, potential CD and controls. Transcripts of both TSLP isoforms were decreased in active CD mucosa. Furin, which was overexpressed in active CD biopsies, was able to cleave TSLP in vitro. Accordingly, refractory and untreated CD supernatants showed higher TSLP-degrading capacity in comparison to treated CD and control supernatants. In our ex vivo model, both TSLP isoforms significantly downregulated IFN-γ and IL-8 production by untreated CD biopsies. Conclusions Reduced mucosal TSLP expression may contribute to intestinal damage in refractory and untreated CD. Further studies are needed to verify whether restoring TSLP might be therapeutically useful especially in refractory patients with CD.
- Published
- 2015
31. Can probiotics modulate human disease by impacting intestinal barrier function?
- Author
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Annick Mercenier, Peter A. Bron, Michiel Kleerebezem, Clara Lucia Garcia-Rodenas, Jerry M. Wells, Robert-Jan M. Brummer, Thomas T. MacDonald, Patrice D. Cani, and UCL - SSS/LDRI - Louvain Drug Research Institute
- Subjects
0301 basic medicine ,Medicine (miscellaneous) ,Gut microbiota ,Gut flora ,Inflammatory bowel disease ,Microbiology ,law.invention ,Gastrointestinal disorders ,03 medical and health sciences ,Probiotic ,law ,Gut barrier ,medicine ,Animals ,Humans ,Host-Microbe Interactomics ,Barrier function ,Irritable bowel syndrome ,VLAG ,Enterocolitis ,Crohn's disease ,Nutrition and Dietetics ,biology ,Probiotics ,Immunity ,Full Papers ,medicine.disease ,biology.organism_classification ,Ulcerative colitis ,3. Good health ,Intestines ,Intestinal Diseases ,030104 developmental biology ,Immunology ,WIAS ,medicine.symptom - Abstract
Intestinal barrier integrity is a prerequisite for homeostasis of mucosal function, which is balanced to maximise absorptive capacity, while maintaining efficient defensive reactions against chemical and microbial challenges. Evidence is mounting that disruption of epithelial barrier integrity is one of the major aetiological factors associated with several gastrointestinal diseases, including infection by pathogens, obesity and diabetes, necrotising enterocolitis, irritable bowel syndrome and inflammatory bowel disease. The notion that specific probiotic bacterial strains can affect barrier integrity fuelled research in whichin vitrocell lines, animal models and clinical trials are used to assess whether probiotics can revert the diseased state back to homeostasis and health. This review catalogues and categorises the lines of evidence available in literature for the role of probiotics in epithelial integrity and, consequently, their beneficial effect for the reduction of gastrointestinal disease symptoms.
- Published
- 2017
32. Matrix Metalloproteinases
- Author
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Paolo Biancheri and Thomas T. MacDonald
- Published
- 2017
33. Th17-type cytokines, IL-6 and TNF-α synergistically activate STAT3 and NF-kB to promote colorectal cancer cell growth
- Author
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Giovanni Monteleone, Massimo Fantini, V. De Simone, Carmine Stolfi, Francesco Pallone, Alfredo Colantoni, Thomas T. MacDonald, Giuseppe S. Sica, Pierpaolo Sileri, D. Di Fusco, G. Ronchetti, Eleonora Franzè, De Simone, V, Franze, E, Ronchetti, G, Colantoni, A, Fantini, Mc, Di Fusco, D, Sica, G, Sileri, P, Macdonald, Tt, Pallone, F, Monteleone, G, and Stolfi, C
- Subjects
STAT3 Transcription Factor ,Cancer Research ,medicine.medical_treatment ,Mice, Transgenic ,Biology ,Settore MED/12 ,Mice ,HT29 Cells ,Genetics ,Molecular Biology ,Immune system ,medicine ,Animals ,Humans ,STAT3 ,Cells, Cultured ,Cell Proliferation ,Interleukin-6 ,Tumor Necrosis Factor-alpha ,Cell growth ,Settore BIO/12 ,Interleukins ,Interleukin-17 ,NF-kappa B ,Natural killer T cell ,Gene Expression Regulation, Neoplastic ,Settore MED/18 - Chirurgia Generale ,Cytokine ,Immunology ,Cancer research ,biology.protein ,Cytokines ,Th17 Cells ,Female ,Original Article ,Tumor necrosis factor alpha ,Interleukin 17 ,Colorectal Neoplasms - Abstract
Colorectal cancers (CRCs) often show a dense infiltrate of cytokine-producing immune/inflammatory cells. The exact contribution of each immune cell subset and cytokine in the activation of the intracellular pathways sustaining CRC cell growth is not understood. Herein, we isolate tumor-infiltrating leukocytes (TILs) and lamina propria mononuclear cells (LPMCs) from the tumor area and the macroscopically unaffected, adjacent, colonic mucosa of patients who underwent resection for sporadic CRC and show that the culture supernatants of TILs, but not of LPMCs, potently enhance the growth of human CRC cell lines through the activation of the oncogenic transcription factors signal transducer and activator of transcription 3 (STAT3) and nuclear factor-kappa B (NF-kB). Characterization of immune cell complexity of TILs and LPMCs reveals no differences in the percentages of T cells, natural killer T cells, natural killer (NK) cells, macrophages and B cells. However, T cells from TILs show a functional switch compared with those from LPMCs to produce large amounts of T helper type 17 (Th17)-related cytokines (that is, interleukin-17A (IL-17A), IL-17F, IL-21 and IL-22), tumor necrosis factor-alpha (TNF-alpha) and IL-6. Individual neutralization of IL-17A, IL-17F, IL-21, IL-22, TNF-alpha or IL-6 does not change TIL-derived supernatant-driven STAT3 and NF-kB activation, as well as their proproliferative effect in CRC cells. In contrast, simultaneous neutralization of both IL-17A and TNF-alpha, which abrogates NF-kB signaling, and IL-22 and IL-6, which abrogates STAT3 signaling, reduces the mitogenic effect of supernatants in CRC cells. IL-17A, IL-21, IL-22, TNF-alpha and IL-6 are also produced in excess in the early colonic lesions in a mouse model of sporadic CRC, associated with enhanced STAT3/NF-kB activation. Mice therapeutically given BP-1-102, an orally bioavailable compound targeting STAT3/NF-kB activation and cross-talk, exhibit reduced colon tumorigenesis and diminished expression of STAT3/NF-kB-activating cytokines in the neoplastic areas. These data suggest that strategies aimed at the cotargeting of STAT3/NF-kB activation and interaction between them might represent an attractive and novel approach to combat CRC.
- Published
- 2014
34. A CD3-Specific Antibody Reduces Cytokine Production and Alters Phosphoprotein Profiles in Intestinal Tissues From Patients With Inflammatory Bowel Disease
- Author
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Kevin R. Page, Lisa Das, Thomas T. MacDonald, Anna Vossenkämper, Christian Hundsrucker, André van Maurik, Andrew J. Stagg, and Theodore J. Sanders
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Adult ,Male ,Chemokine ,Adolescent ,CD3 Complex ,Colon ,Biopsy ,medicine.medical_treatment ,Inflammation ,Biology ,Antibodies, Monoclonal, Humanized ,Proinflammatory cytokine ,Interferon-gamma ,Young Adult ,Intestinal mucosa ,Immune Tolerance ,medicine ,Humans ,Intestinal Mucosa ,Hepatology ,Interleukin-17 ,Gastroenterology ,Antibodies, Monoclonal ,Middle Aged ,Otelixizumab ,Inflammatory Bowel Diseases ,Phosphoproteins ,Interleukin-10 ,Interleukin 10 ,Cytokine ,Case-Control Studies ,Immunology ,biology.protein ,Cytokines ,Female ,Interleukin 17 ,medicine.symptom ,medicine.drug - Abstract
Background & Aims T cells mediate the development of inflammation in inflammatory bowel disease (IBD). We investigated the effects of an antibody against CD3 called otelixizumab, which induces immune tolerance, in intestinal mucosa samples from patients. Methods Intestinal tissues were isolated from patients undergoing routine endoscopy or from patients undergoing intestinal surgery for colon cancer or IBD; healthy surrounding tissues were collected as controls. Isolated lamina propria mononuclear cells (LPMCs) and mucosal tissue explants were incubated with otelixizumab for 24 or 48 hours. Production of inflammatory cytokines was determined by enzyme-linked immunosorbent assay. Levels of 36 cytokines and chemokines and phosphorylation of 39 receptor tyrosine kinases and signaling molecules were measured using protein arrays. Immunoblot analysis was used to analyze T-cell transcription factors. Results Incubation of intestinal tissues or LPMCs with otelixizumab reduced production of interferon gamma, interleukin (IL)-17A, and other inflammatory cytokines and chemokines, simultaneously increasing production of IL-10. Mucosal biopsy specimens from patients with IBD retained inflammation-associated tyrosine phosphoprotein profiles ex vivo. Incubation of the inflamed tissue with otelixizumab reduced phosphorylation of these proteins to levels observed in control tissues. Otelixizumab also markedly reduced phosphorylation of proteins associated with T-cell receptor activation. Neutralization of IL-10 blocked the anti-inflammatory effects of otelixizumab. Conclusions We observed anti-inflammatory effects of anti-CD3 in inflamed intestinal tissues from patients with IBD. The antibody appears to down-regulate T-cell activation via IL-10.
- Published
- 2014
35. Recent advances in gut immunology
- Author
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Thomas T. MacDonald and Nick Powell
- Subjects
0301 basic medicine ,Immunology ,Biology ,03 medical and health sciences ,0302 clinical medicine ,Immune system ,Animals ,Homeostasis ,Humans ,Lymphocytes ,Intestinal Mucosa ,Innate immune system ,Gastrointestinal Microbiome ,Innate lymphoid cell ,Mononuclear phagocyte system ,Acquired immune system ,Intestinal epithelium ,Immunity, Innate ,Gastrointestinal Tract ,Intestines ,Crosstalk (biology) ,030104 developmental biology ,030220 oncology & carcinogenesis ,Parasitology - Abstract
In recent years, there have been significant advances in our understanding of the mucosal immune system. In addition to unravelling some of the complexities of this system, including the discovery of completely new cells types, further insights into the three-way interactions between mucosal immune cells, the intestinal epithelium and the microbial communities colonizing the GI tract promise to redefine our understanding of how intestinal homeostasis is maintained, but also how dysregulation of these highly integrated interactions conspires to cause disease. In this review, we will discuss major recent advances in the role of key immune players in the gut, including innate lymphoid cells (ILCs), mucosa-associated invariant T cells (MAIT cells) and cells of the mononuclear phagocyte system (MPS), including how these cells interact with the intestinal epithelial and their crosstalk with components of the intestinal microbiota, and how these interactions shape host health.
- Published
- 2016
36. P668 Measured and modelled data suggest that oral administration of V565, a novel domain antibody to TNF-alpha, could be beneficial in the treatment of IBD
- Author
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Thomas T. MacDonald, S Nurbhai, M West, S Crowe, Peter M. Irving, Anna Vossenkaemper, and Gareth Parkes
- Subjects
biology ,business.industry ,Gastroenterology ,General Medicine ,Pharmacology ,030226 pharmacology & pharmacy ,Domain (software engineering) ,03 medical and health sciences ,0302 clinical medicine ,Oral administration ,030220 oncology & carcinogenesis ,biology.protein ,Medicine ,Tumor necrosis factor alpha ,Antibody ,business - Published
- 2018
37. Proinflammatory Vδ2+ T Cells Populate the Human Intestinal Mucosa and Enhance IFN-γ Production by Colonic αβ T Cells
- Author
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Shaumick Bhattacharjee, Charlotte Hedin, Theodore J. Sanders, James O. Lindsay, Neil E. McCarthy, Sabrina G. Brown, Anna Vossenkämper, Kevin Whelan, Andrew J. Stagg, Zora Bashir, Thomas T. MacDonald, and Edward M. Giles
- Subjects
medicine.medical_treatment ,Immunology ,Flow cytometry ,Proinflammatory cytokine ,Interferon-gamma ,Interleukin 21 ,Intestinal mucosa ,T-Lymphocyte Subsets ,Addressin ,medicine ,Humans ,Immunology and Allergy ,IL-2 receptor ,Intestinal Mucosa ,Microscopy, Confocal ,biology ,medicine.diagnostic_test ,Receptors, Antigen, T-Cell, gamma-delta ,Flow Cytometry ,Molecular biology ,Phenotype ,Cytokine ,biology.protein ,Lymphocyte Culture Test, Mixed ,Cell activation - Abstract
In nonhuman primates, Vγ9Vδ2+ (Vδ2)T cells proliferate and accumulate in mucosal tissues following microbial activation. Human Vδ2T cells produce proinflammatory cytokines in response to bacterial species that colonize the gut, but the role played by Vδ2T cells in intestinal immunity is unknown. We hypothesized that circulating Vδ2T cells can populate the human intestine and contribute to mucosal inflammation. Cell suspensions prepared from peripheral blood and intestinal biopsies were stimulated with microbial phosphoantigen (1-hydroxy-2-methyl-2-buten-4-yl 4-diphosphate [HDMAPP]) and analyzed by flow cytometry to determine Vδ2T cell phenotype, cytokine production, and proliferative potential. Circulating Vδ2T cells expressed gut-homing integrin α4β7 (>70%), which was coexpressed with skin-associated cutaneous leukocyte Ag by up to 15% of the total population. However, Vδ2T cell activation with HDMAPP and exposure to retinoic acid (signaling via retinoic acid receptor α) increased α4β7 expression and enhanced binding to mucosal addressin cell adhesion molecule-1 in vitro while simultaneously suppressing cutaneous leukocyte Ag, thereby generating a committed gut-tropic phenotype. Confocal microscopy and flow cytometry identified frequent Vδ2T cells that migrated out of human intestinal biopsies and comprised both CD103+ and CD103− subsets that produced TNF-α and IFN-γ upon phosphoantigen exposure, with more frequent cytokine-producing cells in the CD103− population. Activated intestinal Vδ2T cells expressed CD70 and HLA-DR but were unable to drive the proliferation of allogeneic naive CD4+ T cells. Instead, phosphoantigen-activated CD103− Vδ2T cells increased T-bet expression and enhanced IFN-γ production by autologous colonic αβ T cells via an IFN-γ–dependent mechanism. These data demonstrate that circulating Vδ2T cells display enhanced gut-homing potential upon microbial activation and populate the human intestinal mucosa, generating functionally distinct CD103+ and CD103− subsets that can promote inflammation by colonic αβ T cells.
- Published
- 2013
38. IL-25 prevents and cures fulminant hepatitis in mice through a myeloid-derived suppressor cell-dependent mechanism
- Author
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Maria Laura Cupi, Alessandra Gambacurta, Flavio Caprioli, Thomas T. MacDonald, Giovanni Monteleone, Roberta Bernardini, Angelamaria Rizzo, Francesco Pallone, Maurizio Mattei, Eleonora Franzè, Marco Maggioni, Marco Ranalli, G. Ronchetti, Ivan Monteleone, Alfredo Colantoni, and Massimiliano Sarra
- Subjects
Lipopolysaccharides ,Male ,Lipopolysaccharide ,T-Lymphocytes ,Galactosamine ,Hepatitis ,Mice ,chemistry.chemical_compound ,Animals ,Coculture Techniques ,Hepatocytes ,Humans ,Interleukins ,Disease Models, Animal ,Mice, Inbred BALB C ,Cell Proliferation ,Receptors, Chemokine ,Drug-Induced Liver Injury ,Concanavalin A ,Down-Regulation ,Myeloid Cells ,Interleukin-17 ,Antigens, CD11b ,Receptors ,Inbred BALB C ,Settore MED/12 - Gastroenterologia ,CD11b Antigen ,biology ,medicine.diagnostic_test ,Interleukin ,Chemokine ,Chemical and Drug Induced Liver Injury ,Antibody ,medicine.medical_specialty ,Peripheral blood mononuclear cell ,Flow cytometry ,Internal medicine ,medicine ,Antigens ,Hepatology ,Animal ,CD11b ,Molecular biology ,Endocrinology ,chemistry ,Disease Models ,biology.protein ,Myeloid-derived Suppressor Cell ,Liver function - Abstract
Fulminant hepatitis (FH) is a disease characterized by massive destruction of hepatocytes with severe impairment of liver function. The pathogenesis of FH is not fully understood, but hyperactivity of T cells and macrophages with excessive production of cytokines are important hallmarks of the condition. In this study, we investigated the role of interleukin (IL)−25 in FH. IL-25 expression was evaluated in patients with FH and in livers of mice with FH induced by D-galactosamine (D-Gal) and lipopolysaccharide (LPS). Mice were treated with IL-25 before D-Gal/LPS-induced FH and before or after concanavalin A (ConA)-induced FH. Mononuclear cells were isolated from livers of mice treated with or without IL-25 and analyzed for GR1+CD11b+ cells. CFSE-labeled T cells were cocultured with GR1+CD11b+ cells and their proliferation was evaluated by flow cytometry. Mice were also treated with a depleting anti-GR1 antibody before IL-25 and D-Gal/LPS administration. IL-25 was constitutively expressed in mouse and human liver and down-regulated during FH. IL-25 prevented D-Gal/LPS-induced FH and this effect was associated with increased infiltration of the liver with cells coexpressing GR1 and CD11b. In vitro studies showed that GR1+CD11b+ cells isolated from mice given IL-25 inhibited T-cell proliferation. Consistently, in vivo depletion of GR1+ cells abrogated the protective effect of IL-25 in experimental D-Gal/LPS-induced FH. IL-25 was both preventive and therapeutic in ConA-induced FH. Conclusions: IL-25 expression is markedly reduced during human and experimental FH. IL-25 promotes liver accumulation of GR1+CD11b+cells with immunoregulatory properties. (Hepatology 2013;58:1436–1450)
- Published
- 2013
39. Cryptogenic multifocal ulcerating stenosing enteritis associated with homozygous deletion mutations in cytosolic phospholipase A2-α
- Author
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Nicholas S. Kirkby, Franz Rüschendorf, Matthew A. Brooke, Hilary Longhurst, David P. Kelsell, Vincent Plagnol, Thomas T. MacDonald, Jane A. Mitchell, and Timothy D. Warner
- Subjects
Adult ,Genetic Markers ,Male ,Peptic Ulcer ,Sequence analysis ,Blotting, Western ,Fluorescent Antibody Technique ,Gene mutation ,Polymorphism, Single Nucleotide ,Frameshift mutation ,symbols.namesake ,Exon ,Humans ,Frameshift Mutation ,Gene ,Sequence Deletion ,Sanger sequencing ,Genetics ,biology ,Group IV Phospholipases A2 ,Siblings ,Homozygote ,Gastroenterology ,Sequence Analysis, DNA ,Disease gene identification ,PLA2G4A ,Molecular biology ,Enteritis ,Codon, Nonsense ,Case-Control Studies ,biology.protein ,symbols ,Female ,Biomarkers ,Intestinal Obstruction - Abstract
Objective Cryptogenic multifocal ulcerating stenosing enteritis (CMUSE) is an extremely rare, but devastating, disease of unknown aetiology. We investigated the genetic basis of this autosomal recessive condition in a pair of affected siblings who have 40-year histories of catastrophic gastrointestinal and extraintestinal disease. Design Genome-wide single-nucleotide polymorphism homozygosity mapping in the two affected family members combined with whole-exome sequencing of one affected sibling. This was followed by confirmatory Sanger sequencing of the likely disease-causing sequence variant and functional studies in affected and unaffected family members. Results Insertion/deletion variation analysis revealed the presence of a homozygous 4 bp deletion (g.155574_77delGTAA) in the PLA2G4A gene, located in the splice donor site directly after exon 17 (the penultimate exon) of the gene in both affected siblings. This introduces a frameshift of 10 amino acids before a premature stop codon (p.V707fsX10), which is predicted to result in the loss of 43 amino acids (residues 707–749) at the C-terminus of cytosolic phospholipase A2-α (cPLA 2 α). cPLA 2 α protein expression was undetectable in the gut of both siblings, with platelet aggregation and thromboxane A 2 production, as functional assays for cPLA 2 α activity, grossly impaired. Conclusions We have identified mutations in PLA2G4A as a cause of CMUSE in two affected siblings. Further studies are needed to determine if mutations in this gene are also responsible for disease of a similar phenotype in other cases.
- Published
- 2012
40. Neutrophils in ulcerative colitis: a review of selected biomarkers and their potential therapeutic implications
- Author
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Mark Berner Hansen, Daniel Muthas, Clare A. Balendran, Thomas T. MacDonald, Mohib Uddin, Tomas Ottosson, Gerhard Böttcher, Ib Groth Clausen, Carina Kärrman Mårdh, Anna Reznichenko, Silvio Danese, Muthas, D, Reznichenko, A, Balendran, Ca, Bottcher, G, Clausen, Ig, Mardh, Ck, Ottosson, T, Uddin, M, Macdonald, Tt, Danese, S, and Hansen, Mb
- Subjects
0301 basic medicine ,Neutrophils ,Inflammation ,03 medical and health sciences ,Feces ,0302 clinical medicine ,medicine ,Humans ,Molecular Targeted Therapy ,Intestinal Mucosa ,biology ,business.industry ,Lactoferrin ,Gastroenterology ,medicine.disease ,Ulcerative colitis ,Review article ,030104 developmental biology ,C-Reactive Protein ,Immunology ,biology.protein ,030211 gastroenterology & hepatology ,Colitis, Ulcerative ,medicine.symptom ,Calprotectin ,business ,Crypt Abscess ,Leukocyte L1 Antigen Complex ,Elafin ,Biomarkers ,SLPI - Abstract
Objectives: This review article describes the role of neutrophils in mucosal injury and the resulting crypt abscesses characteristic of ulcerative colitis. We also review selected biomarkers for monitoring neutrophil presence and activity in the mucosa as well as their potential as therapeutic targets. Material: We have collated and selectively reviewed data on the most prominent well-established and emerging neutrophil-related biomarkers and potential therapeutic targets (calprotectin, lactoferrin, CXCR1, CXCR2, MMP-9, NGAL, elafin, HNE, pANCAs, MPO, CD16, CD177, CD64, HNPs, SLPI and PTX3) in ulcerative colitis. Results: Systemic and intestinal neutrophil activity increases substantially in active ulcerative colitis, driving tissue damage and extra-intestinal manifestations. Calprotectin is a robust neutrophil and disease biomarker, and a few neutrophil-related targets are being clinically explored as therapeutic targets. Conclusion: We propose that targeting neutrophils and their inflammatory mediators per se is an opportunity that should be explored to identify new effective medical therapies. The overall clinical goal for neutrophil-targeted therapy will be to modulate, but not completely silence, neutrophil activity, thereby abolishing the destructive inflammation with associated acute and chronic tissue damage without compromising host-defense.
- Published
- 2016
41. Critical role for tumor necrosis factor alpha in controlling the number of lumenal pathogenic bacteria and immunopathology in infectious colitis
- Author
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Gad Frankel, Thomas T. MacDonald, Cameron P. Simmons, David J. M. Lewis, Gordon Dougan, Marjan Ghaem-Maghami, G. Monteleone, and Nathalie S. Gonçalves
- Subjects
CD4-Positive T-Lymphocytes ,Male ,Colon ,Immunology ,Gene Expression ,Colonic Diseases, Functional ,CD8-Positive T-Lymphocytes ,Biology ,Infectious Colitis ,Microbiology ,Inflammatory bowel disease ,Receptors, Tumor Necrosis Factor ,Mice ,Immune system ,Antigens, CD ,Immunity ,Immunopathology ,medicine ,Citrobacter rodentium ,Animals ,Intestinal Mucosa ,Colitis ,Mice, Knockout ,Host Response and Inflammation ,Hyperplasia ,Tumor Necrosis Factor-alpha ,Enterobacteriaceae Infections ,STAT4 Transcription Factor ,medicine.disease ,Interleukin-12 ,Citrobacter freundii ,DNA-Binding Proteins ,Mice, Inbred C57BL ,Infectious Diseases ,Receptors, Tumor Necrosis Factor, Type I ,Trans-Activators ,Female ,Parasitology ,Tumor necrosis factor alpha ,Interleukin-4 - Abstract
Infection of mice with the intestinal bacterial pathogenCitrobacter rodentiumresults in colonic mucosal hyperplasia and a local Th1 inflammatory response similar to that seen in mouse models of inflammatory bowel disease. In these latter models, and in patients with Crohn's disease, neutralization of tumor necrosis factor alpha (TNF-α) is of therapeutic benefit. Since there is no information on the role of TNF-α in either immunity to noninvasive bacterial pathogens or on the role of TNF-α in the immunopathology of infectious colitis, we investigatedC. rodentiuminfection in TNFRp55−/−mice. In TNFRp55−/−mice, there were higher colonic bacterial burdens, but the organisms were cleared at the same rate as C57BL/6 mice, showing that TNF-α is not needed for protective antibacterial immunity. The most striking feature of infection in TNFRp55−/−mice, however, was the markedly enhanced pathology, with increased mucosal weight and thickness, increased T-cell infiltrate, and a markedly greater mucosal Th1 response. Interleukin-12 p40 transcripts were markedly elevated inC. rodentium-infected TNFRp55−/−mice, and this was associated with enhanced mucosal STAT4 phosphorylation. TNF-α is not obligatory for protective immunity toC. rodentiumin mice; however, it appears to play some role in downregulating mucosal pathology and Th1 immune responses.
- Published
- 2016
42. Impaired resistance and enhanced pathology during infection with a noninvasive, attaching-effacing enteric bacterial pathogen, Citrobacter rodentium, in mice lacking IL-12 or IFN-gamma
- Author
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Mona Bajaj-Elliott, Bianca C. Neves, Cameron P. Simmons, Marjan Ghaem-Maghami, Gad Frankel, Thomas T. MacDonald, Simon Clare, Gordon Dougan, and Nathalie S. Gonçalves
- Subjects
Male ,Pathology ,medicine.medical_specialty ,beta-Defensins ,Colon ,Immunology ,Colony Count, Microbial ,Administration, Oral ,Nitric Oxide Synthase Type II ,digestive system ,Inflammatory bowel disease ,Bacterial Adhesion ,Microbiology ,Nitric oxide ,Colonic Diseases ,Interferon-gamma ,Mice ,chemistry.chemical_compound ,Immune system ,medicine ,Citrobacter rodentium ,Animals ,Immunology and Allergy ,RNA, Messenger ,Intestinal Mucosa ,Pathogen ,Gene knockout ,Mice, Knockout ,Citrobacter ,Antigens, Bacterial ,biology ,Enterobacteriaceae Infections ,medicine.disease ,biology.organism_classification ,Antibodies, Bacterial ,Interleukin-12 ,Citrobacter freundii ,Mice, Inbred C57BL ,Kinetics ,chemistry ,Interleukin 12 ,Female ,Nitric Oxide Synthase - Abstract
Mice infected with Citrobacter rodentium represent an excellent model in which to examine immune defenses against an attaching-effacing enteric bacterial pathogen. Colonic tissue from mice infected with C. rodentium harbors increased transcripts for IL-12 and IFN-γ and displays mucosal pathology compared with uninfected controls. In this study, the role of IL-12 and IFN-γ in host defense and mucosal injury during C. rodentium infection was examined using gene knockout mice. IL-12p40−/− and IFN-γ−/− mice were significantly more susceptible to mucosal and gut-derived systemic C. rodentium infection. In particular, a proportion of IL-12p40−/− mice died during infection. Analysis of the gut mucosa of IL-12p40−/− mice revealed an influx of CD4+ T cells and a local IFN-γ response. Infected IL-12p40−/− and IFN-γ−/− mice also mounted anti-Citrobacter serum and gut-associated IgA responses and strongly expressed inducible NO synthase (iNOS) in mucosal tissue, despite diminished serum nitrite/nitrate levels. However, iNOS does not detectably contribute to host defense against C. rodentium, as iNOS−/− mice were not more susceptible to infection. However, C57BL/6 mice infected with C. rodentium up-regulated expression of the mouse β-defensin (mBD)-1 and mBD-3 in colonic tissue. In contrast, expression of mBD-3, but not mBD-1, was significantly attenuated during infection of IL-12- and IFN-γ-deficient mice, suggesting mBD-3 may contribute to host defense. These studies are among the first to examine mechanisms of host resistance to an attaching-effacing pathogen and show an important role for IL-12 and IFN-γ in limiting bacterial infection of the colonic epithelium.
- Published
- 2016
43. Effect of narrow spectrum versus selective kinase inhibitors on the intestinal proinflammatory immune tesponse in ulcerative colitis
- Author
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Thomas T. MacDonald, Claire A. Walshe, Paolo Biancheri, Yemisi Solanke, Eleanor Wood, Matthew C. Fyfe, Martyn R Foster, Adele Rowley, Steve Webber, and Sameer Sirohi
- Subjects
0301 basic medicine ,MAPK/ERK pathway ,Niacinamide ,medicine.drug_class ,Biopsy ,Primary Cell Culture ,Dasatinib ,Original Basic Science Articles ,Syk ,Pharmacology ,Naphthalenes ,Fostamatinib ,Mitogen-Activated Protein Kinase 14 ,03 medical and health sciences ,0302 clinical medicine ,medicine ,Immunology and Allergy ,Humans ,Syk Kinase ,Protein kinase A ,Myofibroblasts ,narrow spectrum kinase inhibitor ,Protein Kinase Inhibitors ,ulcerative colitis ,business.industry ,Kinase ,Macrophages ,Interleukin-8 ,TOP1210 ,Gastroenterology ,Protein kinase inhibitor ,030104 developmental biology ,Pyrimidines ,src-Family Kinases ,Benzamides ,Leukocytes, Mononuclear ,Cytokines ,Pyrazoles ,030211 gastroenterology & hepatology ,Colitis, Ulcerative ,Signal transduction ,Janus kinase ,business ,HT29 Cells ,medicine.drug - Abstract
Article first published online 22 April 2016., Background: Kinases are key mediators of inflammation, highlighting the potential of kinase inhibitors as treatments for inflammatory disorders. Selective kinase inhibitors, however, have proved disappointing, particularly in the treatment of rheumatoid arthritis and inflammatory bowel disease. Consequently, to improve efficacy, attention has turned to multikinase inhibition. Methods: The activity of a narrow spectrum kinase inhibitor, TOP1210, has been compared with selective kinase inhibitors (BIRB-796, dasatinib and BAY-61-3606) in a range of kinase assays, inflammatory cell assays, and in inflamed biopsies from patients with ulcerative colitis (UC). Effects on recombinant P38α, Src, and Syk kinase activities were assessed using Z-lyte assays (Invitrogen, Paisley, United Kingdom). Anti-inflammatory effects were assessed by measurement of proinflammatory cytokine release from peripheral blood mononuclear cells, primary macrophages, HT29 cells, inflamed colonic UC biopsies, and myofibroblasts isolated from inflamed colonic UC mucosa. Results: TOP1210 potently inhibits P38α, Src, and Syk kinase activities. Similarly, TOP1210 demonstrates potent inhibitory activity against proinflammatory cytokine release in each of the cellular assays and the inflamed colonic UC biopsies and myofibroblasts isolated from inflamed colonic UC mucosa. Generally, the selective kinase inhibitors showed limited and weaker activity in the cellular assays compared with the broad inhibitory profile of TOP1210. However, combination of the selective inhibitors led to improved efficacy and potency in both cellular and UC biopsy assays. Conclusions: Targeted, multikinase inhibition with TOP1210 leads to a broad efficacy profile in both the innate and adaptive immune responses, with significant advantages over existing selective kinase approaches, and potentially offers a much improved therapeutic benefit in inflammatory bowel disease.
- Published
- 2016
44. Human intestinal barrier function in health and disease
- Author
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Julia König, Annick Mercenier, Clara Lucia Garcia-Rodenas, Jerry M. Wells, Freddy J. Troost, Robert-Jan M. Brummer, Thomas T. MacDonald, Jacqueline Whyte, Patrice D. Cani, RS: FHML non-thematic output, RS: NUTRIM - R2 - Gut-liver homeostasis, Interne Geneeskunde, and UCL - SSS/LDRI - Louvain Drug Research Institute
- Subjects
0301 basic medicine ,Clinical Review ,medicine.medical_specialty ,Gastrointestinal tract ,Allergy ,Intestinal permeability ,business.industry ,Gastroenterology ,Gluten sensitivity ,Disease ,medicine.disease ,Inflammatory bowel disease ,03 medical and health sciences ,030104 developmental biology ,Internal medicine ,Immunology ,medicine ,WIAS ,Life Science ,Host-Microbe Interactomics ,business ,Irritable bowel syndrome ,Barrier function ,VLAG - Abstract
The gastrointestinal tract consists of an enormous surface area that is optimized to efficiently absorb nutrients, water, and electrolytes from food. At the same time, it needs to provide a tight barrier against the ingress of harmful substances, and protect against a reaction to omnipresent harmless compounds. A dysfunctional intestinal barrier is associated with various diseases and disorders. In this review, the role of intestinal permeability in common disorders such as infections with intestinal pathogens, inflammatory bowel disease, irritable bowel syndrome, obesity, celiac disease, non-celiac gluten sensitivity, and food allergies will be discussed. In addition, the effect of the frequently prescribed drugs proton pump inhibitors and non-steroidal anti-inflammatory drugs on intestinal permeability, as well as commonly used methods to assess barrier function will be reviewed.
- Published
- 2016
45. The Transcription Factor T-bet Regulates Intestinal Inflammation Mediated by Interleukin-7 Receptor+ Innate Lymphoid Cells
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Graham M. Lord, Michael A. Curtis, Julian Parkhill, Thomas T. MacDonald, M. Refik Gökmen, Ellen Marks, Richard G. Jenner, Jane K. Howard, Emilie Stolarczyk, Ian Jackson, Nick Powell, Ahmed Hashim, James B. Canavan, and Alan W. Walker
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Immunology ,Biology ,Article ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Immune system ,Immunity ,Helicobacter ,medicine ,Animals ,Immunology and Allergy ,Lymphocytes ,Colitis ,skin and connective tissue diseases ,Interleukin-7 receptor ,Transcription factor ,Cells, Cultured ,030304 developmental biology ,Mice, Knockout ,Mice, Inbred BALB C ,0303 health sciences ,Receptors, Interleukin-7 ,Innate immune system ,Innate lymphoid cell ,medicine.disease ,Immunity, Innate ,3. Good health ,Cell biology ,DNA-Binding Proteins ,body regions ,Infectious Diseases ,1107 Immunology ,Chronic Disease ,Colitis, Ulcerative ,Signal transduction ,T-Box Domain Proteins ,Signal Transduction ,030215 immunology - Abstract
Summary Mice lacking the transcription factor T-bet in the innate immune system develop microbiota-dependent colitis. Here, we show that interleukin-17A (IL-17A)-producing IL-7Rα+ innate lymphoid cells (ILCs) were potent promoters of disease in Tbx21−/−Rag2−/− ulcerative colitis (TRUC) mice. TNF-α produced by CD103−CD11b+ dendritic cells synergized with IL-23 to drive IL-17A production by ILCs, demonstrating a previously unrecognized layer of cellular crosstalk between dendritic cells and ILCs. We have identified Helicobacter typhlonius as a key disease trigger driving excess TNF-α production and promoting colitis in TRUC mice. Crucially, T-bet also suppressed the expression of IL-7R, a key molecule involved in controlling intestinal ILC homeostasis. The importance of IL-7R signaling in TRUC disease was highlighted by the dramatic reduction in intestinal ILCs and attenuated colitis following IL-7R blockade. Taken together, these data demonstrate the mechanism by which T-bet regulates the complex interplay between mucosal dendritic cells, ILCs, and the intestinal microbiota., Highlights ► Chronic colitis in TRUC mice was mediated by IL-17-producing innate lymphoid cells ► TNF-α synergized with IL-23 to induce innate IL-17 production ► Helicobacter typhlonius triggered intestinal pathology in TRUC mice ► T-bet regulated IL-7R transcription, a key checkpoint in intestinal ILC homeostasis
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- 2012
46. Proteases and the gut barrier
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Thomas T. MacDonald, Paolo Biancheri, Antonio Di Sabatino, and Gino Roberto Corazza
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Lamina propria ,Proteases ,Histology ,biology ,Proteolytic enzymes ,Cell Biology ,Matrix metalloproteinase ,Extracellular Matrix ,Pathology and Forensic Medicine ,Cell biology ,Intestines ,Fibronectin ,Extracellular matrix ,medicine.anatomical_structure ,Intestinal mucosa ,Biochemistry ,Laminin ,biology.protein ,medicine ,Animals ,Humans ,Intestinal Mucosa ,Peptide Hydrolases - Abstract
Serine proteases, cysteine proteases, aspartic proteases and matrix metalloproteinases play an essential role in extracellular matrix remodeling and turnover through their proteolytic action on collagens, proteoglycans, fibronectin, elastin and laminin. Proteases can also act on chemokines, receptors and anti-microbial peptides, often potentiating their activity. The intestinal mucosa is the largest interface between the external environment and the tissues of the human body and is constantly exposed to proteolytic enzymes from many sources, including bacteria in the intestinal lumen, fibroblasts and immune cells in the lamina propria and enterocytes. Controlled proteolytic activity is crucial for the maintenance of gut immune homeostasis, for normal tissue turnover and for the integrity of the gut barrier. However, in intestinal immune-mediated disorders, pro-inflammatory cytokines induce the up-regulation of proteases, which become the end-stage effectors of mucosal damage by destroying the epithelium and basement membrane integrity and degrading the extracellular matrix of the lamina propria to produce ulcers. Protease-mediated barrier disruption in turn results in increased amounts of antigen crossing into the lamina propria, driving further immune responses and sustaining the inflammatory process.
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- 2012
47. A rapid diagnostic test for human regulatory T-cell function to enable regulatory T-cell therapy
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Henrieta Fazekasova, Thomas T. MacDonald, Behdad Afzali, Graham M. Lord, Francis C. Edozie, Cristiano Scottà, Maria P. Hernandez-Fuentes, Giovanna Lombardi, and James B. Canavan
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Antigens, Differentiation, T-Lymphocyte ,CD4-Positive T-Lymphocytes ,Time Factors ,Regulatory T cell ,CD40 Ligand ,Immunology ,Population ,Succinimides ,chemical and pharmacologic phenomena ,Immunologic Tests ,Biology ,T-Lymphocytes, Regulatory ,Biochemistry ,Article ,Autoimmune Diseases ,Immune tolerance ,Cell therapy ,Antigens, CD ,Immune Tolerance ,medicine ,Humans ,Lectins, C-Type ,IL-2 receptor ,CD154 ,education ,Cells, Cultured ,Cell Proliferation ,education.field_of_study ,Reproducibility of Results ,FOXP3 ,hemic and immune systems ,Cell Biology ,Hematology ,Flow Cytometry ,Fluoresceins ,Coculture Techniques ,medicine.anatomical_structure ,Cytokines ,Ex vivo - Abstract
Regulatory T cells (CD4+CD25hiCD127loFOXP3+ T cells [Tregs]) are a population of lymphocytes involved in the maintenance of self-tolerance. Abnormalities in function or number of Tregs are a feature of autoimmune diseases in humans. The ability to expand functional Tregs ex vivo makes them ideal candidates for autologous cell therapy to treat human autoimmune diseases and to induce tolerance to transplants. Current tests of Treg function typically take up to 120 hours, a kinetic disadvantage as clinical trials of Tregs will be critically dependent on the availability of rapid diagnostic tests before infusion into humans. Here we evaluate a 7-hour flow cytometric assay for assessing Treg function, using suppression of the activation markers CD69 and CD154 on responder T cells (CD4+CD25− [Tresp]), compared with traditional assays involving inhibition of CFSE dilution and cytokine production. In both freshly isolated and ex vivo expanded Tregs, we describe excellent correlation with gold standard suppressor cell assays. We propose that the kinetic advantage of the new assay may place it as the preferred rapid diagnostic test for the evaluation of Treg function in forthcoming clinical trials of cell therapy, enabling the translation of the large body of preclinical data into potentially useful treatments for human diseases.
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- 2012
48. The aryl hydrocarbon receptor in inflammatory bowel disease: linking the environment to disease pathogenesis
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Ivan Monteleone, Francesco Pallone, Giovanni Monteleone, and Thomas T. MacDonald
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Inflammation ,Inflammatory bowel disease ,Proinflammatory cytokine ,03 medical and health sciences ,0302 clinical medicine ,Immune system ,medicine ,Animals ,Humans ,Intestinal Mucosa ,Immunity, Mucosal ,030304 developmental biology ,0303 health sciences ,Settore MED/12 - Gastroenterologia ,Innate immune system ,biology ,Gastroenterology ,Environmental exposure ,Environmental Exposure ,respiratory system ,Aryl hydrocarbon receptor ,medicine.disease ,Inflammatory Bowel Diseases ,3. Good health ,Disease Models, Animal ,Receptors, Aryl Hydrocarbon ,Immunology ,biology.protein ,Intraepithelial lymphocyte ,030211 gastroenterology & hepatology ,medicine.symptom - Abstract
Purpose of review The aryl hydrocarbon receptor (AhR), a transcription factor activated by a large number of environmental agents, modulates the activity of immune and nonimmune cells in the gut, and may represent an important link between the environment and the immune perturbations which underlie the pathogenesis of inflammatory bowel disease. This review will summarize the current knowledge of the role of AhR in regulation of intestinal immune homeostasis and inflammation. Recent findings Activation of AhR by dietary ligands is necessary for the maintenance or expansion of innate immune cells in the gut, such as intraepithelial lymphocytes (IELs) and interleukin (IL)-22-producing lymphoid cells (ILC22). AhR-deficient mice lack IELs, have reduced number of ILC22 cells, and are more susceptible to bacterial infections and experimental colitis. In animal models, AhR activators inhibit proinflammatory cytokine synthesis and attenuate colitis by a pathway that involves IL-22. Analysis of AhR in the human gut reveals that intestinal T cells and natural killer cells isolated from Crohn's disease patients express low levels of AhR and respond to AhR ligands by downregulating inflammatory cytokines and upregulating IL-22. Summary These novel findings may help explain how environmental factors may regulate mucosal immune responses.
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- 2012
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49. Recent advances in understanding ulcerative colitis
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Antonio Di Sabatino, Thomas T. MacDonald, Laura Rovedatti, Paolo Biancheri, and Gino Roberto Corazza
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Male ,medicine.medical_specialty ,Toxic megacolon ,Colorectal cancer ,Rectum ,Risk Assessment ,Severity of Illness Index ,Gastroenterology ,Inflammatory bowel disease ,Megacolon, Toxic ,chemistry.chemical_compound ,Mesalazine ,Recurrence ,Internal medicine ,Internal Medicine ,medicine ,Humans ,Intestinal Mucosa ,Colitis ,Mesalamine ,Colectomy ,Megacolon ,business.industry ,Biopsy, Needle ,Prognosis ,medicine.disease ,Combined Modality Therapy ,Immunohistochemistry ,Ulcerative colitis ,digestive system diseases ,Survival Rate ,medicine.anatomical_structure ,chemistry ,Disease Progression ,Emergency Medicine ,Colitis, Ulcerative ,Female ,Colorectal Neoplasms ,business ,Precancerous Conditions - Abstract
Ulcerative colitis, one of the two main forms of inflammatory bowel disease, is characterized by inflammation of the large bowel with constant involvement of the rectum, and a possible continuous retrograde distribution up to the cecum. Typical macroscopic lesions are mucosal ulcerations, with immune cell infiltration and cryptic abscesses at histology. Ulcerative colitis usually manifests with bloody diarrhea, is associated with a number of extra-intestinal manifestations, and may be acutely complicated by toxic megacolon. Longstanding disease may predispose to the development of colorectal cancer. Therapeutic options include mesalazine, corticosteroids, immunomodulators and biologic agents; however, if these treatments fail, the only available therapeutic choice remaining is the surgical removal of the colon. This review emphasizes novel concepts in the basic aspects of ulcerative colitis, and, in addition to the current clinical and diagnostic knowledge, it also describes new treatment options for this condition.
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- 2011
50. The endogenous cannabinoid system in the gut of patients with inflammatory bowel disease
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Giuseppe Astarita, Paolo Biancheri, Alessandro Vanoli, Enrico Dainese, Laura Rovedatti, Thomas T. MacDonald, Mauro Maccarrone, Daniele Piomelli, Marco Guerci, Cinzia Rapino, Sylvia L.F. Pender, Gino Roberto Corazza, Natalia Battista, and A. Di Sabatino
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Cannabinoid receptor ,medicine.medical_treatment ,Immunology ,Arachidonic Acids ,Pharmacology ,Biology ,Inflammatory bowel disease ,Proinflammatory cytokine ,Receptor, Cannabinoid, CB2 ,Mice ,chemistry.chemical_compound ,Receptor, Cannabinoid, CB1 ,Fatty acid amide hydrolase ,Cannabinoid Receptor Modulators ,medicine ,Animals ,Humans ,Immunology and Allergy ,Intestinal Mucosa ,Myofibroblasts ,Methanandamide ,Anandamide ,STAT4 Transcription Factor ,Inflammatory Bowel Diseases ,medicine.disease ,Endocannabinoid system ,Intestines ,chemistry ,Cytokines ,lipids (amino acids, peptides, and proteins) ,Cannabinoid ,T-Box Domain Proteins - Abstract
Activation of cannabinoid receptors (CBs) by endocannabinoids impacts on a number of gastrointestinal functions. Recent data indicate that CB1 agonists improve 2,4-dinitrobenzene sulfonic acid-induced colitis in mice, thus suggesting a role for the endocannabinoid agonist anandamide (AEA) in protecting the gut against inflammation. We here examined the gut endocannabinoid system in inflammatory bowel disease (IBD) patients, and investigated the ex vivo and in vitro effects of the non-hydrolysable AEA analog methanandamide (MAEA) on the mucosal proinflammatory response. The content of AEA, but not of 2-arachidonoyl-glycerol and N-palmitoylethanolamine, was significantly lower in inflamed than uninflamed IBD mucosa, and this was paralleled by lower activity of the AEA-synthesizing enzyme N-acyl-phosphatidylethanolamine-specific phospholipase D and higher activity of the AEA-degrading enzyme fatty acid amide hydrolase. MAEA significantly downregulated interferon-γ and tumor necrosis factor-α secretion by both organ culture biopsies and lamina propria mononuclear cells. Although these results are promising, further studies are needed to determine the role of cannabinoid pathways in gut inflammation. © 2011 Society for Mucosal Immunology.
- Published
- 2011
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