Functional and molecular analysis of the epithelial barrier of porcine Peyer's patches follicle-associated epithelium

• Applicant: Professor Dr. Salah Amasheh
• Subject Area: Veterinary Medical Science
• Term: from 2016 to 2024
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 325849034

• Final Report Year: 2025

Final Report Abstract

The majority of immune cells are located in the gastrointestinal tract, demonstrating that the gut-associated lymphoid tissue (GALT) plays a pivotal role in the immune defense of the body. The GALT is composed of single isolated lymphoid follicles and aggregated follicles called Peyer´s patches (PP) The epithelium in this specific location is named follicleassociated epithelium (FAE) and contains specialized microfold cells (M cells), which have the function of presenting antigens to the underlying lymph follicles by transcytosis to induce an immune response or tolerance against the antigens. Therefore, the paracellular barrier in this region is tighter to ensure the route through the M cells to prevent an uncontrolled uptake of antigens. The capability of the immune system to balance between potentially harmful and helpful immune response is crucial to prevent an overactive immune system that could lead to tissue damage. This complex regulation is disturbed in inflammatory bowel diseases (IBDs) such as Crohn´s disease and ulcerative colitis. The pro inflammatory cytokine Tumor necrosis factor alpha (TNF) and its receptors TNF receptor 1 (TNFR-1) and TNF receptor 2 (TNFR-2) play a crucial role in the disease progression in IBDs. This project aimed to characterize barrier determinants in porcine PP, neighboring tissue and cell models, namely tight junction proteins, and the effects of endogenous and exogenous effectors. Our project provided detailed insights in the effects of TNF, as well as the effects of beneficially, mainly secondary plant compounds including cannabidiol, berberine, quercetin, cinnamic acid, and caffeic acid on barrier integrity and regulation. During the funding period, 12 peer review original publications and a series of conference abstracts and presentations, and two promotions of doctoral students emerged.

Publications

• Analysis of the pH-dependent interaction of human gastric TJ proteins expressed in Xenopus laevis oocytes. 4th International Tight Junction Conference Berlin – 27.09.-29.09.2021
  Stein, L.; Brunner, N. & Amasheh S.
  
• Effects of 1,2-Dimethylhydrazine on Barrier Properties of Rat Large Intestine and IPEC-J2 Cells. International Journal of Molecular Sciences, 22(19), 10278.
  Bekusova, Viktoria; Droessler, Linda; Amasheh, Salah & Markov, Alexander G.
  
• Exploring the function of the gastric barrier protein Claudin-18 in a heterologuous expression model. 75. digitale Tagung der Gesellschaft für Ernährungsphysiologie Berlin – 16.03.-18.03.2021. DLG-Verlag GmbH. Proceedings of the Society of Nutrition Physiology : Berichte der Gesellschaft für Ernährungsphysiologie; volume 30, S. 110. ISBN: 978-3-7690-4114-9
  Stein, L.; Brunner, N. & Amasheh S.
  
• TNFa induces epithelial barrier perturbation by a self-enhancing effect on TNFR-1 in porcine jejunal epithelial cells. 100th annual meeting of the German Physiological Society: book of abstracts: 30.09.– 02.10.2021 Frankfurt am Main – DPG, Deutsche Physiologische Gesellschaft (Hrsg.), S. 450
  Droessler, L.; Cornelius, V.; Markov, A. & Amasheh S.
  
• TNFα induced changes in the composition of tight junction proteins in follicle-associated epithelium of porcine Peyer`s Patches. 75. digitale Tagung der Gesellschaft für Ernährungsphysiologie Berlin – 16.03.-18.03.2021. DLG-Verlag GmbH. Proceedings of the Society of Nutrition Physiology: Berichte der Gesellschaft für Ernährungsphysiologie; volume 30 (2021), S. 105 ISBN: 978-3-7690-4114-9
  Droessler, L.; Cornelius, V. & Amasheh S.
  
• TNFα-effects on tight junction proteins in IPEC-J2 cells are associated with changes in TNFR-1 and MLCK. 4th International Tight Junction Conference Berlin – 27.09.-29.09.2021
  Drößler, L.; Cornelius, V. & Amasheh S.
  
• Tumor Necrosis Factor Alpha Effects on the Porcine Intestinal Epithelial Barrier Include Enhanced Expression of TNF Receptor 1. International Journal of Molecular Sciences, 22(16), 8746.
  Droessler, Linda; Cornelius, Valeria; Markov, Alexander G. & Amasheh, Salah
  
• Analysis of cannabidiol effects on the epithelial barrier of por-cine intestinal IPEC-J2 cells. Tagung der Fachgruppe Physiologie und Biochemie der Deutschen Veterinärmedizinischen Gesellschaft (DVG) Gießen – 24.06.-26.06.2022
  Böhm, E.; Drößler, L. & Amasheh S.
  
• Analysis of ZO-1 in claudin-expressing Xenopus laevis oocytes. Tagung der Fachgruppe Physiologie und Biochemie der Deutschen Veterinärmedizinischen Gesellschaft (DVG) Gießen – 24.06.-26.06.2022
  Brunner, N.; Stein, L. & Amasheh S.
  
• Barrier Perturbation in Porcine Peyer’s Patches by Tumor Necrosis Factor is Associated With a Dysregulation of Claudins. Frontiers in Physiology, 13.
  Droessler, Linda; Cornelius, Valeria; Boehm, Elisa; Stein, Laura; Brunner, Nora & Amasheh, Salah
  
• Cellular Distribution Pattern of tjp1 (ZO-1) in Xenopus laevis Oocytes Heterologously Expressing Claudins. The Journal of Membrane Biology, 256(1), 51-61.
  Brunner, Nora; Stein, Laura & Amasheh, Salah
  
• Concerted action of berberine in the porcine intestinal epithelial model IPEC‐J2: Effects on tight junctions and apoptosis. Physiological Reports, 10(7).
  Cornelius, Valeria; Droessler, Linda; Boehm, Elisa & Amasheh, Salah
  
• Functional Analysis of Gastric Tight Junction Proteins in Xenopus laevis Oocytes. Membranes, 12(8), 731.
  Stein, Laura; Brunner, Nora & Amasheh, Salah
  
• Selective Role of TNFα and IL10 in Regulation of Barrier Properties of the Colon in DMH-Induced Tumor and Healthy Rats. International Journal of Molecular Sciences, 23(24), 15610.
  Bekusova, Viktoria; Zudova, Tatiana; Fatyykhov, Ilyas; Fedorova, Arina; Amasheh, Salah & Markov, Alexander G.
  
• Cannabidiol attenuates inflammatory impairment of intestinal cells expanding biomaterial-based therapeutic approaches. Materials Today Bio, 23, 100808.
  Boehm, Elisa; Droessler, Linda & Amasheh, Salah
  
• Cannabidiol effects on gastroin-testinal tight junction proteins. 102th annual meeting of the German Physiological Society Berlin – 21.09.-23.09.2023. 102th annual meeting of the German Physiological Society : book of abstracts – Deutsche Physiologische Gesellschaft (Hrsg.) S. 170–171
  Boehm, E.; Vollstaedt, M. L.; Stein, L.; Droessler, L. & Amasheh S.
  
• CBD prevents TNF-induced barrier disturbance in in-testinal epithelial cells. 2nd Microphysiological Systems World Summit Berlin – 26.06.-30.06.2023. In: ALTEX proceedings; 11(1), S. 51. (2023)
  Boehm E.; Drößler L.; Trappe S. & Amasheh S.
  
• Effects of TNF and Cannabidiol on porcine intestinal IPEC-J2 cells. 77. Tagung der Gesellschaft für Ernährungsphysiologie Göttingen – 07.03.-09.03.2023. Proceedings of the Society of Nutrition Physiology – GfE (Hrsg.) Frankfurt: DLG Verlag; 32, S. 85 ISBN: 978-3-7690-4116-3
  Boehm E.; Drößler L.; Trappe S. & Amasheh S.
  
• Prednisolone Targets Claudins in Mouse Brain Blood Vessels. International Journal of Molecular Sciences, 25(1), 276.
  Markov, Alexander G.; Bikmurzina, Anastasia E.; Fedorova, Arina A.; Vinogradova, Ekaterina P.; Kruglova, Natalia M.; Krivoi, Igor I. & Amasheh, Salah
  
• Cannabidiol Strengthening of Gastric Tight Junction Complexes Analyzed in an Improved Xenopus Oocyte Assay. Membranes, 14(1), 18.
  Stein, Laura; Vollstaedt, Marie-Luise & Amasheh, Salah
  
• Cinnamic Acid and Caffeic Acid Effects on Gastric Tight Junction Proteins Analyzed in Xenopus laevis Oocytes. Membranes, 14(2), 40.
  Vollstädt, Marie-Luise; Stein, Laura; Brunner, Nora & Amasheh, Salah
  
• Quercetin Improves Barrier Properties in Porcine Small Intestine but Not in Peyer’s Patches. International Journal of Molecular Sciences, 25(3), 1530.
  Cornelius, Valeria; Droessler, Linda & Amasheh, Salah