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

Gaelle Noel; Nicholas W. Baetz; Janet F. Staab; Mark Donowitz; Olga Kovbasnjuk; Marcela F. Pasetti; Nicholas C. Zachos

doi:10.1038/srep46790

A Primary Human Macrophage-Enteroid Co-Culture Model to Investigate Mucosal Gut Physiology and Host-Pathogen Interactions

Gastroenterology ◽

10.1016/s0016-5085(17)30545-0 ◽

2017 ◽

Vol 152 (5) ◽

pp. S56-S57

Author(s):

Nicholas W. Baetz ◽

Gaelle Noel ◽

Janet Staab ◽

Mark Donowitz ◽

Olga Kovbasnjuk ◽

...

Keyword(s):

Human Macrophage ◽

Primary Human Macrophage ◽

Host Pathogen Interactions ◽

Culture Model ◽

Host Pathogen ◽

Gut Physiology

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

Scientific Reports ◽

10.1038/srep45270 ◽

2017 ◽

Vol 7 (1) ◽

Cited By ~ 114

Author(s):

Gaelle Noel ◽

Nicholas W. Baetz ◽

Janet F. Staab ◽

Mark Donowitz ◽

Olga Kovbasnjuk ◽

...

Keyword(s):

Intestinal Epithelium ◽

Immune Cells ◽

Barrier Function ◽

Morphological Changes ◽

Human Monocyte ◽

Human Macrophage ◽

Primary Human Macrophage ◽

Cell Height ◽

Culture Model ◽

Gut Physiology

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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Lipidomic Profiling of Host‐Pathogen Interactions Using Untargeted and Targeted Lipidomics Approaches in a Sepsis Cell Culture Model

The FASEB Journal ◽

10.1096/fasebj.2020.34.s1.04970 ◽

2020 ◽

Vol 34 (S1) ◽

pp. 1-1

Author(s):

Kaushalya Amunugama ◽

David A. Ford

Keyword(s):

Cell Culture ◽

Cell Culture Model ◽

Host Pathogen Interactions ◽

Culture Model ◽

Host Pathogen

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Application of a gut-immune co-culture system for the study of N-glycan-dependent host-pathogen interactions of Campylobacter jejuni

10.1101/702472 ◽

2019 ◽

Author(s):

Cristina Y. Zamora ◽

Elizabeth M. Ward ◽

Jemila C. Kester ◽

Wen Li Kelly Chen ◽

Jason G. Velazquez ◽

...

Keyword(s):

Immune System ◽

Campylobacter Jejuni ◽

Outer Membrane Vesicles ◽

Protein Glycosylation ◽

Bacterial Invasion ◽

Host Pathogen Interactions ◽

Barrier Functions ◽

Immune System Activation ◽

Culture Model ◽

Host Pathogen

AbstractAn in vitro gut-immune co-culture model with apical and basal accessibility, designed to more closely resemble a human intestinal microenvironment, was employed to study the role of the Nlinked protein glycosylation (Pgl) pathway in Campylobacter jejuni pathogenicity. The gutimmune co-culture (GIC) was developed to model important aspects of the human small intestine by the inclusion of mucin producing goblet cells, human enterocytes, and dendritic cells, bringing together a mucus-containing epithelial monolayer with elements of the innate immune system. The utility of the system was demonstrated by characterizing host-pathogen interactions facilitated by N-linked glycosylation, such as host epithelial barrier functions, bacterial invasion and immunogenicity. Changes in human intestinal barrier functions in the presence of 11168 C. jejuni (wildtype) strains were quantified using GICs. The glycosylationdeficient strain 11168 ΔpglE was 100-fold less capable of adhering to and invading this intestinal model in cell infectivity assays. Quantification of inflammatory signaling revealed that 11168ΔpglE differentially modulated inflammatory responses in different intestinal microenvironments, suppressive in some but activating in others. Virulence-associated outer membrane vesicles produced by wildtype and 11168ΔpglE C. jejuni were shown to have differential composition and function, with both leading to immune system activation when provided to the gut-immune co-culture model. This analysis of aspects of C. jejuni infectivity in the presence and absence of its N-linked glycome, is enabled by application of the gut-immune model and we anticipate that this system will be applicable to further studies of C. jejuni and other enteropathogens of interest.

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A novel 3D culture model of fungal keratitis to explore host-pathogen interactions within the stromal environment

Experimental Eye Research ◽

10.1016/j.exer.2021.108581 ◽

2021 ◽

Vol 207 ◽

pp. 108581

Author(s):

Marina E. Brown ◽

Micaela L. Montgomery ◽

Manali M. Kamath ◽

Sarah Nicholas ◽

Yutao Liu ◽

...

Keyword(s):

3D Culture ◽

Fungal Keratitis ◽

Host Pathogen Interactions ◽

Culture Model ◽

Host Pathogen ◽

3D Culture Model

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Application of a gut-immune co-culture system for the study of N-glycan-dependent host–pathogen interactions of Campylobacter jejuni

Glycobiology ◽

10.1093/glycob/cwz105 ◽

2020 ◽

Vol 30 (6) ◽

pp. 374-381 ◽

Cited By ~ 1

Author(s):

Cristina Y Zamora ◽

Elizabeth M Ward ◽

Jemila C Kester ◽

Wen Li Kelly Chen ◽

Jason G Velazquez ◽

...

Keyword(s):

Immune System ◽

Campylobacter Jejuni ◽

Outer Membrane Vesicles ◽

Protein Glycosylation ◽

Bacterial Invasion ◽

Host Pathogen Interactions ◽

Barrier Functions ◽

Immune System Activation ◽

Culture Model ◽

Host Pathogen

Abstract An in vitro gut-immune co-culture model with apical and basal accessibility, designed to more closely resemble a human intestinal microenvironment, was employed to study the role of the N-linked protein glycosylation pathway in Campylobacter jejuni pathogenicity. The gut-immune co-culture (GIC) was developed to model important aspects of the human small intestine by the inclusion of mucin-producing goblet cells, human enterocytes and dendritic cells, bringing together a mucus-containing epithelial monolayer with elements of the innate immune system. The utility of the system was demonstrated by characterizing host–pathogen interactions facilitated by N-linked glycosylation, such as host epithelial barrier functions, bacterial invasion and immunogenicity. Changes in human intestinal barrier functions in the presence of 11168 C. jejuni (wildtype) strains were quantified using GICs. The glycosylation-impaired strain 11168 ΔpglE was 100-fold less capable of adhering to and invading this intestinal model in cell infectivity assays. Quantification of inflammatory signaling revealed that 11168ΔpglE differentially modulated inflammatory responses in different intestinal microenvironments, suppressive in some but activating in others. Virulence-associated outer membrane vesicles produced by wildtype and 11168ΔpglE C. jejuni were shown to have differential composition and function, with both leading to immune system activation when provided to the gut-immune co-culture model. This analysis of aspects of C. jejuni infectivity in the presence and absence of its N-linked glycome is enabled by application of the gut-immune model, and we anticipate that this system will be applicable to further studies of C. jejuni and other enteropathogens of interest.

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A co-culture model of the bovine alveolus

F1000Research ◽

10.12688/f1000research.18696.2 ◽

2019 ◽

Vol 8 ◽

pp. 357 ◽

Cited By ~ 2

Author(s):

Diane Lee ◽

Mark Chambers

Keyword(s):

Epithelial Cells ◽

Alveolar Type ◽

Type Ii Cells ◽

Type Ii ◽

Host Pathogen Interactions ◽

Alveolar Type Ii Cells ◽

Culture Model ◽

Host Pathogen

The epithelial lining of the lung is often the first point of interaction between the host and inhaled pathogens, allergens and medications. Epithelial cells are therefore the main focus of studies which aim to shed light on host-pathogen interactions, to dissect the mechanisms of local host immunity and study toxicology. If these studies are not to be conducted exclusively in vivo, it is imperative that in vitro models are developed with a high in vitro- in vivo correlation. We describe here a co-culture model of the bovine alveolus, designed to overcome some of the limitations encountered with mono-culture and live animal models. Our system includes bovine pulmonary arterial endothelial cells (BPAECs) seeded onto a permeable membrane in 24 well Transwell format. The BPAECs are overlaid with immortalised bovine alveolar type II epithelial cells and cultured at air-liquid interface for 14 days before use; in our case to study host-mycobacterial interactions. Characterisation of novel cell lines and the co-culture model have provided compelling evidence that immortalised bovine alveolar type II cells are an authentic substitute for primary alveolar type II cells and their co-culture with BPAECs provides a physiologically relevant in vitro model of the bovine alveolus. The co-culture model may be used to study dynamic intracellular and extracellular host-pathogen interactions, using proteomics, genomics, live cell imaging, in-cell ELISA and confocal microscopy. The model presented in this article enables other researchers to establish an in vitro model of the bovine alveolus that is easy to set up, malleable and serves as a comparable alternative to in vivo models, whilst allowing study of early host-pathogen interactions, currently not feasible in vivo. The model therefore achieves one of the 3Rs objectives in that it replaces the use of animals in research of bovine respiratory diseases.

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