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Cited by 420Gut microbiota and healthEscherichia coli research studiesGastrointestinal motility and disorders

A primary human macrophage-enteroid co-culture model to investigate mucosal gut physiology and host-pathogen interactions

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Noel, Gaelle, Baetz, Nicholas W., Staab, Janet F., Donowitz, Mark, Kovbasnjuk, Olga, Pasetti, Marcela F., Zachos, Nicholas C.·2017·Scientific Reports
DOI 10.1038/srep45270PMC PMC5366908
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Abstract

Integration of the intestinal epithelium and the mucosal immune system is critical for gut homeostasis. The intestinal epithelium is a functional barrier that secludes luminal content, senses changes in the gut microenvironment, and releases immune regulators that signal underlying immune cells. However, interactions between epithelial and innate immune cells to maintain barrier integrity and prevent infection are complex and poorly understood. We developed and characterized a primary human macrophage-enteroid co-culture model for in-depth studies of epithelial and macrophage interactions. Human intestinal stem cell-derived enteroid monolayers co-cultured with human monocyte-derived macrophages were used to evaluate barrier function, cytokine secretion, and protein expression under basal conditions and following bacterial infection. Macrophages enhanced barrier function and maturity of enteroid monolayers as indicated by increased transepithelial electrical resistance and cell height. Communication between the epithelium and macrophages was demonstrated through morphological changes and cytokine production. Intraepithelial macrophage projections, efficient phagocytosis, and stabilized enteroid barrier function revealed a coordinated response to enterotoxigenic and enteropathogenic E. coli infections. In summary, we have established the first primary human macrophage-enteroid co-culture system, defined conditions that allow for a practical and reproducible culture model, and demonstrated its suitability to study gut physiology and host responses to enteric pathogens.

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  • Human intestinal organoid-derived monolayers have been successfully used to study host-pathogen interactions, epithelial barrier function, and various transport processes including secretion of hormones, growth factors, and ions, and absorption of glucose.

    “Human intestinal organoid-derived monolayers have been successfully used to study host–pathogen interactions [10,11,12], epithelial barrier function [13], and various transport processes, including secretion of hormones, growth factors, and ions [14,15,16], and absorption of glucose [17].”

    From Regulation of Enterocyte Brush Border Membrane Primary Na-Absorptive Transporters in Human Intestinal Organoid-Derived Monolayers (2024) · cites this paper as [12]

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Research works printed but not linked: 51

  1. Peterson L. W. & Artis D. Intestinal epithelial cells: regulators of barrier function and immune homeostasis. Nat Rev Immunol 14, 141– 153, doi: 10.1038/nri3608 (2014). 24566914 · DOI
  2. Duell B. L., Cripps A. W., Schembri M. A. & Ulett G. C. Epithelial Cell Coculture Models for Studying Infectious Diseases: Benefits and Limitations. Journal of Biomedicine and Biotechnology 2011, 852419, doi: 10.1155/2011/852419 (2011). 22007147 PMC3189631 · DOI
  3. Gibbons D. L. & Spencer J. Mouse and human intestinal immunity: same ballpark, different players; different rules, same score. Mucosal Immunol 4, 148– 157 (2011). 21228770
  4. Tsilingiri K.. Probiotic and postbiotic activity in health and disease: comparison on a novel polarised ex-vivo organ culture model. Gut 61, 1007– 1015, doi: 10.1136/gutjnl-2011-300971 (2012). 22301383 · DOI
  5. Bimczok D.. Human gastric epithelial cells contribute to gastric immune regulation by providing retinoic acid to dendritic cells. Mucosal Immunol 8, 533– 544, doi: 10.1038/mi.2014.86 (2015). 25249167 PMC4372513 · DOI
  6. Bisping G.. Patients with inflammatory bowel disease (IBD) reveal increased induction capacity of intracellular interferon-gamma (IFN-gamma) in peripheral CD8+ lymphocytes co-cultured with intestinal epithelial cells. Clin Exp Immunol 123, 15– 22 (2001). 11167992 PMC1905965
  7. Leonard F., Collnot E.-M. & Lehr C.-M. A Three-Dimensional Coculture of Enterocytes, Monocytes and Dendritic Cells To Model Inflamed Intestinal Mucosa in Vitro. Molecular Pharmaceutics 7, 2103– 2119, doi: 10.1021/mp1000795 (2010). 20809575 · DOI
  8. McKay D. M., Croitoru K. & Perdue M. H. T cell-monocyte interactions regulate epithelial physiology in a coculture model of inflammation. Am J Physiol 270, C418– 428 (1996). 8779903
  9. Parkos C. A., Delp C., Arnaout M. A. & Madara J. L. Neutrophil migration across a cultured intestinal epithelium. Dependence on a CD11b/CD18-mediated event and enhanced efficiency in physiological direction. Journal of Clinical Investigation 88, 1605– 1612 (1991). 1682344 PMC295682
  10. Nanthakumar N. N., Fusunyan R. D., Sanderson I. & Walker W. A. Inflammation in the developing human intestine: A possible pathophysiologic contribution to necrotizing enterocolitis. Proc Natl Acad Sci USA 97, 6043– 6048 (2000). 10823949 PMC18555
  11. Trapecar M., Goropevsek A., Gorenjak M., Gradisnik L. & Slak Rupnik M. A Co-Culture Model of the Developing Small Intestine Offers New Insight in the Early Immunomodulation of Enterocytes and Macrophages by italic Lactobacillus spp. italic through STAT1 and NF-kB p65 Translocation. PLoS ONE 9, e86297, doi: 10.1371/journal.pone.0086297 (2014). PMC3894201 24454965 · DOI
  12. Bermudez-Brito M., Plaza-Diaz J., Fontana L., Munoz-Quezada S. & Gil A. In vitro cell and tissue models for studying host-microbe interactions: a review. Br J Nutr 109 Suppl 2, S27– 34, doi: 10.1017/S0007114512004023 (2013). 23360878 · DOI
  13. Foulke-Abel J.. Human Enteroids as a Model of Upper Small Intestinal Ion Transport Physiology and Pathophysiology. Gastroenterology 150, 638– 649 e638, doi: 10.1053/j.gastro.2015.11.047 (2016). 26677983 PMC4766025 · DOI
  14. In J. G.. Human mini-guts: new insights into intestinal physiology and host-pathogen interactions. Nat Rev Gastroenterol Hepatol 13, 633– 642, doi: 10.1038/nrgastro.2016.142 (2016). 27677718 PMC5079760 · DOI · Paper page
  15. Sato T. & Clevers H. Growing self-organizing mini-guts from a single intestinal stem cell: mechanism and applications. Science 340, 1190– 1194, doi: 10.1126/science.1234852 (2013). 23744940 · DOI · Paper page
  16. Zachos N. C.. Human Enteroids/Colonoids and Intestinal Organoids Functionally Recapitulate Normal Intestinal Physiology and Pathophysiology. J Biol Chem 291, 3759– 3766, doi: 10.1074/jbc.R114.635995 (2016). 26677228 PMC4759158 · DOI
  17. Stelzner M.. A nomenclature for intestinal in vitro cultures. Am J Physiol Gastrointest Liver Physiol 302, G1359– 1363, doi: 10.1152/ajpgi.00493.2011 (2012). 22461030 PMC3378093 · DOI · Paper page
  18. Barker N. & Clevers H. Lineage tracing in the intestinal epithelium. Curr Protoc Stem Cell Biol Chapter 5, Unit5A 4, doi: 10.1002/9780470151808.sc05a04s13 (2010). 20443207 · DOI
  19. Mukherjee S. & Hooper L. V. Antimicrobial defense of the intestine. Immunity 42, 28– 39, doi: 10.1016/j.immuni.2014.12.028 (2015). 25607457 · DOI
  20. Durand A.. Functional intestinal stem cells after Paneth cell ablation induced by the loss of transcription factor Math1 (Atoh1). Proc Natl Acad Sci USA 109, 8965– 8970, doi: 10.1073/pnas.1201652109 (2012). 22586121 PMC3384132 · DOI
  21. Smythies L. E.. Mucosal IL-8 and TGF-beta recruit blood monocytes: evidence for cross-talk between the lamina propria stroma and myeloid cells. J Leukoc Biol 80, 492– 499, doi: 10.1189/jlb.1005566 (2006). 16793909 · DOI
  22. Tabbaa M. G., Axon A. T. R. & Dixon M. F. Enterocyte dimensions in patients with abnormal intestinal permeability. European Journal of Gastroenterology and Hepatology 6, 607– 610 (1994).
  23. Grant C. N., Nellis E. D. & Chahine A. A. A case of small intestinal cast causing SBO in complicated intestinal graft-versus-host disease. Pediatr Surg Int 30, 685– 688, doi: 10.1007/s00383-014-3515-9 (2014). 24805116 · DOI
  24. Mizutani T.. Real-time analysis of P-glycoprotein-mediated drug transport across primary intestinal epithelium three-dimensionally cultured in vitro. Biochem Biophys Res Commun 419, 238– 243, doi: 10.1016/j.bbrc.2012.01.155 (2012). 22342245 · DOI
  25. Walker N. M.. Cellular chloride and bicarbonate retention alters intracellular pH regulation in Cftr KO crypt epithelium. Am J Physiol Gastrointest Liver Physiol 310, G70– 80, doi: 10.1152/ajpgi.00236.2015 (2016). 26542396 PMC4719062 · DOI
  26. Spottl T.. A new organotypic model to study cell interactions in the intestinal mucosa. Eur J Gastroenterol Hepatol 18, 901– 909 (2006). 16825910
  27. Carvalho H. M., Teel L. D., Goping G. & O’Brien A. D. A three-dimensional tissue culture model for the study of attach and efface lesion formation by enteropathogenic and enterohaemorrhagic Escherichia coli. Cell Microbiol 7, 1771– 1781, doi: 10.1111/j.1462-5822.2004.00594.x (2005). 16309463 · DOI
  28. Nickerson C. A.. Three-dimensional tissue assemblies: novel models for the study of Salmonella enterica serovar Typhimurium pathogenesis. Infect Immun 69, 7106– 7120, doi: 10.1128/IAI.69.11.7106-7120.2001 (2001). 11598087 PMC100098 · DOI
  29. Barker N. Adult intestinal stem cells: critical drivers of epithelial homeostasis and regeneration. Nat Rev Mol Cell Biol 15, 19– 33, doi: 10.1038/nrm3721 (2014). 24326621 · DOI
  30. Middendorp S.. Adult stem cells in the small intestine are intrinsically programmed with their location-specific function. Stem Cells 32, 1083– 1091, doi: 10.1002/stem.1655 (2014). 24496776 · DOI
  31. VanDussen K. L.. Development of an enhanced human gastrointestinal epithelial culture system to facilitate patient-based assays. Gut 64, 911– 920, doi: 10.1136/gutjnl-2013-306651 (2015). 25007816 PMC4305344 · DOI · Paper page
  32. Smith P. D.. Intestinal macrophages and response to microbial encroachment. Mucosal Immunol 4, 31– 42, doi: 10.1038/mi.2010.66 (2011). 20962772 PMC3821935 · DOI
  33. Smythies L. E.. Inflammation anergy in human intestinal macrophages is due to Smad-induced IkappaBalpha expression and NF-kappaB inactivation. J Biol Chem 285, 19593– 19604, doi: 10.1074/jbc.M109.069955 (2010). 20388715 PMC2885238 · DOI
  34. Italiani P. & Boraschi D. From Monocytes to M1/M2 Macrophages: Phenotypical vs. Functional Differentiation. Front Immunol 5, 514, doi: 10.3389/fimmu.2014.00514 (2014). 25368618 PMC4201108 · DOI · Paper page
  35. Saxena K.. Human Intestinal Enteroids: a New Model To Study Human Rotavirus Infection, Host Restriction, and Pathophysiology. J Virol 90, 43– 56, doi: 10.1128/JVI.01930-15 (2015). 26446608 PMC4702582 · DOI · Paper page
  36. Zhang Y. G., Wu S., Xia Y. & Sun J. Salmonella-infected crypt-derived intestinal organoid culture system for host-bacterial interactions. Physiol Rep 2, doi: 10.14814/phy2.12147 (2014). PMC4270227 25214524 · DOI
  37. Gonzales-Siles L. & Sjoling A. The different ecological niches of enterotoxigenic Escherichia coli. Environ Microbiol 18, 741– 751, doi: 10.1111/1462-2920.13106 (2016). 26522129 PMC4982042 · DOI
  38. Humphries R. M. & Armstrong G. D. Sticky situation: localized adherence of enteropathogenic Escherichia coli to the small intestine epithelium. Future Microbiol 5, 1645– 1661, doi: 10.2217/fmb.10.124 (2010). 21133687 · DOI
  39. Rescigno M., Rotta G., Valzasina B. & Ricciardi-Castagnoli P. Dendritic cells shuttle microbes across gut epithelial monolayers. Immunobiology 204, 572– 581, doi: 10.1078/0171-2985-00094 (2001). 11846220 · DOI
  40. Strisciuglio C.. Impaired autophagy leads to abnormal dendritic cell-epithelial cell interactions. J Crohns Colitis 7, 534– 541, doi: 10.1016/j.crohns.2012.08.009 (2012). 22981596 · DOI
  41. Yin Y.. CpG DNA assists the whole inactivated H9N2 influenza virus in crossing the intestinal epithelial barriers via transepithelial uptake of dendritic cell dendrites. Mucosal Immunol 8, 799– 814, doi: 10.1038/mi.2014.110 (2014). 25492476 · DOI
  42. Chieppa M., Rescigno M., Huang A. Y. & Germain R. N. Dynamic imaging of dendritic cell extension into the small bowel lumen in response to epithelial cell TLR engagement. J Exp Med 203, 2841– 2852, doi: 10.1084/jem.20061884 (2006). 17145958 PMC2118178 · DOI
  43. Mazzini E., Massimiliano L., Penna G. & Rescigno M. Oral tolerance can be established via gap junction transfer of fed antigens from CX3CR1(+) macrophages to CD103(+) dendritic cells. Immunity 40, 248– 261, doi: 10.1016/j.immuni.2013.12.012 (2014). 24462723 · DOI
  44. Travis M. A.. Loss of integrin alpha(v)beta8 on dendritic cells causes autoimmunity and colitis in mice. Nature 449, 361– 365, doi: 10.1038/nature06110 (2007). 17694047 PMC2670239 · DOI · Paper page
  45. Wang Y. P.. Effects of Wnt3a on proliferation, activation and the expression of TGFb/Smad in rat hepatic stellate cells. Zhonghua Gan Zang Bing Za Zhi 21, 111– 115 (2013). 23663882
  46. Shi M.. Latent TGF-beta structure and activation. Nature 474, 343– 349, doi: 10.1038/nature10152 (2011). 21677751 PMC4717672 · DOI · Paper page
  47. Mills M. & Estes M. K. Physiologically relevant human tissue models for infectious diseases. Drug Discov Today 21, 1540– 1552, doi: 10.1016/j.drudis.2016.06.020 (2016). 27352632 PMC5365153 · DOI
  48. Evans D. G., Silver R. P., Evans D. J. Jr., Chase D. G. & Gorbach S. L. Plasmid-controlled colonization factor associated with virulence in Esherichia coli enterotoxigenic for humans. Infect Immun 12, 656– 667 (1975). 1100526 PMC415337
  49. Evans D. J. Jr. & Evans D. G. Three characteristics associated with enterotoxigenic Escherichia coli isolated from man. Infect Immun 8, 322– 328 (1973). 4581006 PMC422851
  50. Levine M. M.. Escherichia coli strains that cause diarrhoea but do not produce heat-labile or heat-stable enterotoxins and are non-invasive. Lancet 1, 1119– 1122 (1978). 77415
  51. Cleary J.. Enteropathogenic Escherichia coli (EPEC) adhesion to intestinal epithelial cells: role of bundle-forming pili (BFP), EspA filaments and intimin. Microbiology 150, 527– 538, doi: 10.1099/mic.0.26740-0 (2004). 14993302 · DOI

Cited by

Sorted by citations

Authors
Noel, G., Baetz, N. W., Staab, J. F., Donowitz, M., Kovbasnjuk, O., Pasetti, M. F., & Zachos, N. C.
Affiliations
  • University of Maryland, Baltimore (US)
  • Johns Hopkins University (US)
Corresponding author
  • Pasetti, Marcela F.
  • Zachos, Nicholas C.
ORCID
Venue
Scientific Reports
Publisher
Springer Science and Business Media LLC
Published
2017-03-27
Volume
7
Issue
1
Pages
45270-45270
ISSN
2045-2322 (ISSN-L)
DOI
10.1038/srep45270
PMC
PMC5366908
OpenAlex
W2603154743
Cited by
420
References
7
Citations by year
2026: 25 · 2025: 44 · 2024: 41 · 2023: 48 · 2022: 56 · 2021: 68 · 2020: 56 · 2019: 41 · 2018: 26 · 2017: 14
FWCI
15.31
Type
journal article
Language
en
Update notices
Open access
Licence
Locations
  • Scientific Reports (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Funders
  • Johns Hopkins University
  • National Institutes of Health (DK089502)
  • National Institute of Allergy and Infectious Diseases (P01AI125181)
  • National Institute of Diabetes and Digestive and Kidney Diseases (P30DK089502)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Gut microbiota and health
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Endocrinology › Escherichia coli research studies
  • Health Sciences › Medicine › Gastroenterology › Gastrointestinal motility and disorders
Keywords
intestinal enteroids, human monocyte-derived macrophages, epithelial-macrophage interaction, intestinal barrier function, transepithelial electrical resistance, enteropathogenic Escherichia coli, enterotoxigenic Escherichia coli, host-pathogen interaction, cytokine production, phagocytosis, intestinal homeostasis, mucosal immunity
MeSH terms
Bacteria, Cell Communication, Cells, Cultured, Epithelial Cells, Humans, Intestinal Mucosa, Macrophages, Models, Biological, Cytokines, Coculture Techniques, Host-Pathogen Interactions

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