Semi-Quantification of Ex-Vivo Sonoelastography in Mesenterial Lymph Nodes from Patients with Adenocarcinomas and Crohn's Disease

Abstract Background and Aims Creeping fat, the wrapping of mesenteric fat around the bowel wall, is a typical feature of Crohn’s disease, and is associated with stricture formation and bowel obstruction. How creeping fat forms is unknown, and we interrogated potential mechanisms using novel intestinal tissue and cell interaction systems. Methods Tissues from normal, ulcerative colitis, non-strictured and strictured Crohn’s disease intestinal specimens were obtained. Fresh and decellularized tissue, mesenteric fat explants, primary human adipocytes, pre-adipocytes, muscularis propria cells, and native extracellular matrix were used in multiple ex vivo and in vitro systems involving cell growth, differentiation and migration, proteomics, and integrin expression. Results Crohn’s disease muscularis propria cells produced an extracellular matrix scaffold which is in direct spatial and functional contact with the immediately overlaid creeping fat. The scaffold contained multiple proteins, but only fibronectin production was singularly upregulated by TGF-b1. The muscle cell-derived matrix triggered migration of pre-adipocytes out of mesenteric fat, fibronectin being the dominant factor responsible for their migration. Blockade of α5β1 on the pre-adipocyte surface inhibited their migration out of mesenteric fat and on 3D decellularized intestinal tissue extracellular matrix. Conclusion Crohn’s disease creeping fat appears to result from the migration of pre-adipocytes out of mesenteric fat and differentiation into adipocytes in response to an increased production of fibronectin by activated muscularis propria cells. These new mechanistic insights may lead to novel approaches for prevention of creeping fat-associated stricture formation.

Download Full-text

Novel strain-level resolution of Crohn’s disease mucosa-associated microbiota via an ex vivo combination of microbe culture and metagenomic sequencing

The ISME Journal ◽

10.1038/s41396-021-00991-1 ◽

2021 ◽

Author(s):

J. J. Teh ◽

E. M. Berendsen ◽

E. C. Hoedt ◽

S. Kang ◽

J. Zhang ◽

...

Keyword(s):

Crohn’S Disease ◽

Crohn's Disease ◽

Ex Vivo ◽

Taxonomic Composition ◽

Strain Level ◽

Patient Specific ◽

Rrna Gene ◽

Metagenomic Sequencing ◽

Functional Characterisation ◽

Bacterial Lineages

AbstractThe mucosa-associated microbiota is widely recognized as a potential trigger for Crohn’s disease pathophysiology but remains largely uncharacterised beyond its taxonomic composition. Unlike stool microbiota, the functional characterisation of these communities using current DNA/RNA sequencing approaches remains constrained by the relatively small microbial density on tissue, and the overwhelming amount of human DNA recovered during sample preparation. Here, we have used a novel ex vivo approach that combines microbe culture from anaerobically preserved tissue with metagenome sequencing (MC-MGS) to reveal patient-specific and strain-level differences among these communities in post-operative Crohn’s disease patients. The 16 S rRNA gene amplicon profiles showed these cultures provide a representative and holistic representation of the mucosa-associated microbiota, and MC-MGS produced both high quality metagenome-assembled genomes of recovered novel bacterial lineages. The MC-MGS approach also produced a strain-level resolution of key Enterobacteriacea and their associated virulence factors and revealed that urease activity underpins a key and diverse metabolic guild in these communities, which was confirmed by culture-based studies with axenic cultures. Collectively, these findings using MC-MGS show that the Crohn’s disease mucosa-associated microbiota possesses taxonomic and functional attributes that are highly individualistic, borne at least in part by novel bacterial lineages not readily isolated or characterised from stool samples using current sequencing approaches.

Download Full-text

Antagonism of Adherent Invasive E. coli LF82 With Human α-defensin 5 in the Follicle-associated Epithelium of Patients With Ileal Crohn’s Disease

Inflammatory Bowel Diseases ◽

10.1093/ibd/izaa315 ◽

2020 ◽

Author(s):

Lina Y Alkaissi ◽

Martin E Winberg ◽

Stéphanie DS Heil ◽

Staffan Haapaniemi ◽

Pär Myrelid ◽

...

Keyword(s):

Crohn’S Disease ◽

Crohn's Disease ◽

Ex Vivo ◽

Ussing Chambers ◽

E Coli ◽

Follicle Associated Epithelium ◽

Vitro Model ◽

Set Up

Abstract Background The first visible signs of Crohn’s disease (CD) are microscopic erosions over the follicle-associated epithelium (FAE). The aim of the study was to investigate the effects of human α-defensin 5 (HD5) on adherent-invasive Escherichia coli LF82 translocation and HD5 secretion after LF82 exposure in an in vitro model of human FAE and in human FAE ex vivo. Methods An in vitro FAE-model was set up by the coculture of Raji B cells and Caco-2-cl1 cells. Ileal FAE from patients with CD and controls were mounted in Ussing chambers. The effect of HD5 on LF82 translocation was studied by LF82 exposure to the cells or tissues with or without incubation with HD5. The HD5 secretion was measured in human FAE exposed to LF82 or Salmonella typhimurium. The HD5 levels were evaluated by immunofluorescence, immunoblotting, and ELISA. Results There was an increased LF82 translocation across the FAE-model compared with Caco-2-cl1 (P < 0.05). Incubation of cell/tissues with HD5 before LF82 exposure reduced bacterial passage in both models. Human FAE showed increased LF82 translocation in CD compared with controls and attenuated passage after incubation with sublethal HD5 in both CD and controls (P < 0.05). LF82 exposure resulted in a lower HD5 secretion in CD FAE compared with controls (P < 0.05), whereas Salmonella exposure caused equal secretion on CD and controls. There were significantly lower HD5 levels in CD tissues compared with controls. Conclusions Sublethal HD5 reduces the ability of LF82 to translocate through FAE. The HD5 is secreted less in CD in response to LF82, despite a normal response to Salmonella. This further implicates the integrated role of antimicrobial factors and barrier function in CD pathogenesis.

Download Full-text

Pathogenic Yersinia DNA Is Detected in Bowel and Mesenteric Lymph Nodes From Patients With Crohn's Disease

The American Journal of Surgical Pathology ◽

10.1097/00000478-200302000-00011 ◽

2003 ◽

Vol 27 (2) ◽

pp. 220-227 ◽

Cited By ~ 73

Author(s):

Laura W. Lamps ◽

K. T. Madhusudhan ◽

Jennifer M. Havens ◽

Joel K. Greenson ◽

Mary P. Bronner ◽

...

Keyword(s):

Crohn’S Disease ◽

Crohn's Disease ◽

Lymph Nodes ◽

Mesenteric Lymph Nodes ◽

Mesenteric Lymph ◽

Pathogenic Yersinia

Download Full-text

2101 Non-Necrotizing Granulomas of the GI Tract and Abdominal Lymph Nodes. Crohn’s Disease, Sarcoidosis or Both?

The American Journal of Gastroenterology ◽

10.14309/01.ajg.0000597936.24257.a6 ◽

2019 ◽

Vol 114 (1) ◽

pp. S1171-S1171

Author(s):

Scott R. Douglas ◽

Nitisha Lotun ◽

Danielle Moore ◽

Amy Doran ◽

R. Ann Hays

Keyword(s):

Crohn’S Disease ◽

Crohn's Disease ◽

Lymph Nodes ◽

Gi Tract ◽

Necrotizing Granulomas

Download Full-text

Differential Pathogenic Th17 Profile in Mesenteric Lymph Nodes of Crohn's Disease and Ulcerative Colitis Patients

Frontiers in Immunology ◽

10.3389/fimmu.2019.01177 ◽

2019 ◽

Vol 10 ◽

Cited By ~ 8

Author(s):

Marwa Bsat ◽

Laurence Chapuy ◽

Manuel Rubio ◽

Ramses Wassef ◽

Carole Richard ◽

...

Keyword(s):

Ulcerative Colitis ◽

Crohn’S Disease ◽

Crohn's Disease ◽

Lymph Nodes ◽

Mesenteric Lymph Nodes ◽

Mesenteric Lymph

Download Full-text

CD4+ Tissue-resident Memory T Cells Expand and Are a Major Source of Mucosal Tumour Necrosis Factor α in Active Crohn’s Disease

Journal of Crohn s and Colitis ◽

10.1093/ecco-jcc/jjz010 ◽

2019 ◽

Vol 13 (7) ◽

pp. 905-915 ◽

Cited By ~ 9

Author(s):

Shrinivas Bishu ◽

Mohammed El Zaatari ◽

Atsushi Hayashi ◽

Guoqing Hou ◽

Nicole Bowers ◽

...

Keyword(s):

T Cells ◽

Crohn’S Disease ◽

T Cell ◽

Crohn's Disease ◽

Tumour Necrosis Factor ◽

Tumour Necrosis ◽

Ex Vivo ◽

Factor Α ◽

And Control ◽

Necrosis Factor

Abstract Background and Aims Tumour necrosis factor [TNF]α- and IL-17A-producing T cells are implicated in Crohn’s disease [CD]. Tissue-resident memory T [TRM] cells are tissue-restricted T cells that are regulated by PR zinc finger domain 1 [PRDM1], which has been implicated in pathogenic Th17 cell responses. TRM cells provide host defence but their role in CD is unknown. We thus examined CD4+ TRM cells in CD. Methods Colon samples were prospectively collected at endoscopy or surgery in CD and control subjects. Flow cytometry and ex vivo assays were performed to characterise CD4+ TRM cells. Results CD4+ TRM cells are the most abundant memory T cell population and are the major T cell source of mucosal TNFα in CD. CD4+ TRM cells are expanded in CD and more avidly produce IL-17A and TNFα relative to control cells. There was a unique population of TNFα+IL-17A+ CD4+ TRM cells in CD which are largely absent in controls. PRDM1 was highly expressed by CD4+ TRM cells but not by other effector T cells. Suppression of PRDM1 was associated with impaired induction of IL17A and TNFA by CD4+ TRM cells Conclusions CD4+ TRM cells are expanded in CD and are a major source of TNFα, suggesting that they are important in CD. PRDM1 is expressed by TRM cells and may regulate their function. Collectively, this argues for prospective studies tracking CD4+ TRM cells over the disease course.

Download Full-text