Differential Effects of TNF (TNFSF2) and IFN-γ on Intestinal Epithelial Cell Morphogenesis and Barrier Function in Three-dimensional Culture

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2011 Aug 20

PMID 21853060

Citations 15

Authors

Kati Juuti-Uusitalo

Leon J Klunder

Klaas A Sjollema

Katarina Mackovicova

Ryuichi Ohgaki

Dick Hoekstra

Jan Dekker

Sven C D van Ijzendoorn

Affiliations

Soon will be listed here.

Abstract

Background: The cytokines TNF (TNFSF2) and IFNγ are important mediators of inflammatory bowel diseases and contribute to enhanced intestinal epithelial permeability by stimulating apoptosis and/or disrupting tight junctions. Apoptosis and tight junctions are also important for epithelial tissue morphogenesis, but the effect of TNF and IFNγ on the process of intestinal epithelial morphogenesis is unknown.

Methods/principal Findings: We have employed a three-dimensional cell culture system, reproducing in vivo-like multicellular organization of intestinal epithelial cells, to study the effect of TNF on intestinal epithelial morphogenesis and permeability. We show that human intestinal epithelial cells in three-dimensional culture assembled into luminal spheres consisting of a single layer of cells with structural, internal, and planar cell polarity. Exposure of preformed luminal spheres to TNF or IFNγ enhanced paracellular permeability, but via distinctive mechanisms. Thus, while both TNF and IFNγ, albeit in a distinguishable manner, induced the displacement of selected tight junction proteins, only TNF increased paracellular permeability via caspase-driven apoptosis and cell shedding. Infliximab and adalumimab inhibited these effects of TNF. Moreover, we demonstrate that TNF via its stimulatory effect on apoptosis fundamentally alters the process of intestinal epithelial morphogenesis, which contributes to the de novo generation of intestinal epithelial monolayers with increased permeability. Also IFNγ contributes to the de novo formation of monolayers with increased permeability, but in a manner that does not involve apoptosis.

Conclusions: Our study provides an optimized 3D model system for the integrated analysis of (real-time) intestinal epithelial paracellular permeability and morphogenesis, and reveals apoptosis as a pivotal mechanism underlying the enhanced permeability and altered morphogenesis in response to TNF, but not IFNγ.

Citing Articles

Understanding the therapeutic toolkit for inflammatory bowel disease.

Vieujean S, Jairath V, Peyrin-Biroulet L, Dubinsky M, Iacucci M, Magro F Nat Rev Gastroenterol Hepatol. 2025; .

PMID: 39891014 DOI: 10.1038/s41575-024-01035-7.

Small nucleolar RNAs and SNHGs in the intestinal mucosal barrier: Emerging insights and current roles.

Yang T, Shen J J Adv Res. 2022; 46:75-85.

PMID: 35700920 PMC: 10105082. DOI: 10.1016/j.jare.2022.06.004.

Regulation of Intestinal Epithelial Barrier and Immune Function by Activated T Cells.

Le N, Mazahery C, Nguyen K, Levine A Cell Mol Gastroenterol Hepatol. 2020; 11(1):55-76.

PMID: 32659380 PMC: 7596298. DOI: 10.1016/j.jcmgh.2020.07.004.

Overexpression of BCL-2 in the Intestinal Epithelium Prevents Sepsis-Induced Gut Barrier Dysfunction via Altering Tight Junction Protein Expression.

Otani S, Oami T, Yoseph B, Klingensmith N, Chen C, Liang Z Shock. 2019; 54(3):330-336.

PMID: 31626040 PMC: 7156322. DOI: 10.1097/SHK.0000000000001463.

The Intestinal Barrier in Parkinson's Disease: Current State of Knowledge.

van Ijzendoorn S, Derkinderen P J Parkinsons Dis. 2019; 9(s2):S323-S329.

PMID: 31561386 PMC: 6839484. DOI: 10.3233/JPD-191707.

References

Capaldo C, Nusrat A . Cytokine regulation of tight junctions. Biochim Biophys Acta. 2008; 1788(4):864-71. PMC: 2699410. DOI: 10.1016/j.bbamem.2008.08.027. View

Debnath J, Muthuswamy S, Brugge J . Morphogenesis and oncogenesis of MCF-10A mammary epithelial acini grown in three-dimensional basement membrane cultures. Methods. 2003; 30(3):256-68. DOI: 10.1016/s1046-2023(03)00032-x. View

Toyli M, Rosberg-Kulha L, Capra J, Vuoristo J, Eskelinen S . Different responses in transformation of MDCK cells in 2D and 3D culture by v-Src as revealed by microarray techniques, RT-PCR and functional assays. Lab Invest. 2010; 90(6):915-28. DOI: 10.1038/labinvest.2010.63. View

Martin-Belmonte F, Yu W, Rodriguez-Fraticelli A, Ewald A, Ewald A, Werb Z . Cell-polarity dynamics controls the mechanism of lumen formation in epithelial morphogenesis. Curr Biol. 2008; 18(7):507-13. PMC: 2405957. DOI: 10.1016/j.cub.2008.02.076. View

Gitter A, Bendfeldt K, Schulzke J, Fromm M . Leaks in the epithelial barrier caused by spontaneous and TNF-alpha-induced single-cell apoptosis. FASEB J. 2000; 14(12):1749-53. DOI: 10.1096/fj.99-0898com. View

Weaver V, Howlett A, Petersen O, Bissell M . The development of a functionally relevant cell culture model of progressive human breast cancer. Semin Cancer Biol. 1995; 6(3):175-84. DOI: 10.1006/scbi.1995.0021. View

Schulzke J, Ploeger S, Amasheh M, Fromm A, Zeissig S, Troeger H . Epithelial tight junctions in intestinal inflammation. Ann N Y Acad Sci. 2009; 1165:294-300. DOI: 10.1111/j.1749-6632.2009.04062.x. View

Radtke F, Clevers H . Self-renewal and cancer of the gut: two sides of a coin. Science. 2005; 307(5717):1904-9. DOI: 10.1126/science.1104815. View

OBrien L, Jou T, Pollack A, Zhang Q, Hansen S, Yurchenco P . Rac1 orientates epithelial apical polarity through effects on basolateral laminin assembly. Nat Cell Biol. 2001; 3(9):831-8. DOI: 10.1038/ncb0901-831. View

10.

Sands B, Kaplan G . The role of TNFalpha in ulcerative colitis. J Clin Pharmacol. 2007; 47(8):930-41. DOI: 10.1177/0091270007301623. View

11.

Amasheh M, Fromm A, Krug S, Amasheh S, Andres S, Zeitz M . TNFalpha-induced and berberine-antagonized tight junction barrier impairment via tyrosine kinase, Akt and NFkappaB signaling. J Cell Sci. 2010; 123(Pt 23):4145-55. DOI: 10.1242/jcs.070896. View

12.

Bruewer M, Luegering A, Kucharzik T, Parkos C, Madara J, Hopkins A . Proinflammatory cytokines disrupt epithelial barrier function by apoptosis-independent mechanisms. J Immunol. 2003; 171(11):6164-72. DOI: 10.4049/jimmunol.171.11.6164. View

13.

Papadakis K, Targan S . Role of cytokines in the pathogenesis of inflammatory bowel disease. Annu Rev Med. 2000; 51:289-98. DOI: 10.1146/annurev.med.51.1.289. View

14.

Jaffe A, Kaji N, Durgan J, Hall A . Cdc42 controls spindle orientation to position the apical surface during epithelial morphogenesis. J Cell Biol. 2008; 183(4):625-33. PMC: 2582895. DOI: 10.1083/jcb.200807121. View

15.

Bruewer M, Samarin S, Nusrat A . Inflammatory bowel disease and the apical junctional complex. Ann N Y Acad Sci. 2006; 1072:242-52. DOI: 10.1196/annals.1326.017. View

16.

Bryant D, Mostov K . From cells to organs: building polarized tissue. Nat Rev Mol Cell Biol. 2008; 9(11):887-901. PMC: 2921794. DOI: 10.1038/nrm2523. View

17.

Fleming E, Zajac M, Moschenross D, Montrose D, Rosenberg D, Cowan A . Planar spindle orientation and asymmetric cytokinesis in the mouse small intestine. J Histochem Cytochem. 2007; 55(11):1173-80. PMC: 3957531. DOI: 10.1369/jhc.7A7234.2007. View

18.

Schock F, Perrimon N . Molecular mechanisms of epithelial morphogenesis. Annu Rev Cell Dev Biol. 2002; 18:463-93. DOI: 10.1146/annurev.cellbio.18.022602.131838. View

19.

Melnick M, Chen H, Zhou Y, Jaskoll T . Embryonic mouse submandibular salivary gland morphogenesis and the TNF/TNF-R1 signal transduction pathway. Anat Rec. 2001; 262(3):318-30. DOI: 10.1002/1097-0185(20010301)262:3<318::AID-AR1023>3.0.CO;2-3. View

20.

Yu W, Shewan A, Brakeman P, Eastburn D, Datta A, Bryant D . Involvement of RhoA, ROCK I and myosin II in inverted orientation of epithelial polarity. EMBO Rep. 2008; 9(9):923-9. PMC: 2529350. DOI: 10.1038/embor.2008.135. View