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Article ; Online: Epithelial Cell Infection Analyses with Shigella.

Poore, Kender / Lenneman, Bryan R / Faherty, Christina S

Journal of visualized experiments : JoVE

2024 , Issue 204

Abstract: The human-adapted enteric bacterial pathogen Shigella causes millions of infections each year, creates long-term growth effects among pediatric patients, and is a leading cause of diarrheal deaths worldwide. Infection induces watery or bloody diarrhea as ...

Abstract	The human-adapted enteric bacterial pathogen Shigella causes millions of infections each year, creates long-term growth effects among pediatric patients, and is a leading cause of diarrheal deaths worldwide. Infection induces watery or bloody diarrhea as a result of the pathogen transiting the gastrointestinal tract and infecting the epithelial cells lining the colon. With staggering increases in antibiotic resistance and the current lack of approved vaccines, standardized research protocols are critical to studying this formidable pathogen. Here, methodologies are presented to examine the molecular pathogenesis of Shigella using in vitro analyses of bacterial adherence, invasion, and intracellular replication in colonic epithelial cells. Prior to infection analyses, the virulence phenotype of Shigella colonies was verified by the uptake of the Congo red dye on agar plates. Supplemented laboratory media can also be considered during bacterial culturing to mimic in vivo conditions. Bacterial cells are then used in a standardized protocol to infect colonic epithelial cells in tissue culture plates at an established multiplicity of infection with adaptations to analyze each stage of infection. For adherence assays, Shigella cells are incubated with reduced media levels to promote bacterial contact with epithelial cells. For both invasion and intracellular replication assays, gentamicin is applied for various time intervals to eliminate extracellular bacteria and enable assessment of invasion and/or the quantification of intracellular replication rates. All infection protocols enumerate adherent, invaded, and/or intracellular bacteria by serially diluting infected epithelial cell lysates and plating bacterial colony forming units relative to infecting titers on Congo red agar plates. Together, these protocols enable independent characterization and comparisons for each stage of Shigella infection of epithelial cells to study this pathogen successfully.
MeSH term(s)	Humans ; Child ; Agar ; Congo Red ; Shigella ; Epithelial Cells ; Dysentery, Bacillary ; Diarrhea
Chemical Substances	Agar (9002-18-0) ; Congo Red (3U05FHG59S)
Language	English
Publishing date	2024-02-09
Publishing country	United States
Document type	Journal Article ; Video-Audio Media
ZDB-ID	2259946-0
ISSN	1940-087X ; 1940-087X
ISSN (online)	1940-087X
ISSN	1940-087X
DOI	10.3791/66426
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Gastrointestinal signals in supplemented media reveal a role in adherence for the

León, Yrvin / Honigsberg, Raphael / Rasko, David A / Faherty, Christina S

Gut microbes

2024 Volume 16, Issue 1, Page(s) 2331985

Abstract: ... Shigella ... ...

Abstract	Shigella flexneri
MeSH term(s)	Type V Secretion Systems/genetics ; Shigella flexneri/genetics ; Gastrointestinal Microbiome ; Epithelial Cells/microbiology ; Mutation ; Escherichia coli ; Bacterial Proteins/genetics
Chemical Substances	Type V Secretion Systems ; Bacterial Proteins
Language	English
Publishing date	2024-03-28
Publishing country	United States
Document type	Journal Article
ZDB-ID	2575755-6
ISSN	1949-0984 ; 1949-0984
ISSN (online)	1949-0984
ISSN	1949-0984
DOI	10.1080/19490976.2024.2331985
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Bacteriophages against enteropathogens: rediscovery and refinement of novel antimicrobial therapeutics.

León, Yrvin / Faherty, Christina S

Current opinion in infectious diseases

2021 Volume 34, Issue 5, Page(s) 491–499

Abstract: Purpose of review: Alarming rates of antibiotic resistance in bacteria and gastrointestinal dysbiosis associated with traditional antimicrobial therapy have led to renewed interests in developing bacteriophages as novel therapeutics. In this review, we ... ...

Abstract	Purpose of review: Alarming rates of antibiotic resistance in bacteria and gastrointestinal dysbiosis associated with traditional antimicrobial therapy have led to renewed interests in developing bacteriophages as novel therapeutics. In this review, we highlight some of the recent advances in bacteriophage therapeutic development targeting important enteropathogens of the gastrointestinal tract. Recent findings: Bacteriophages are viruses that infect bacteria, either to utilize the bacterial machinery to produce new progeny or stably integrate into the bacterial chromosome to ensure maintenance of the viral genome. With recent advances in synthetic biology and the discovery of CRISPR-Cas systems used by bacteria to protect against bacteriophages, novel molecular applications are taking us beyond the discovery of bacteriophages and toward innovative applications, including the targeting of bacterial virulence factors, the use of temperate bacteriophages, and the production of bacteriophage proteins as antimicrobial agents. These technologies offer promise to target enteropathogens without disrupting the healthy microbiota of the gastrointestinal tract. Moreover, the use of nanoparticle technology and other modifications are helping researchers circumvent the harsh gastrointestinal conditions that could limit the efficacy of bacteriophages against enteric pathogens. Summary: This era of discovery and development offers significant potential to modify bacteriophages and overcome the global impact of enteropathogens.
MeSH term(s)	Anti-Bacterial Agents/therapeutic use ; Bacteria ; Bacteriophages ; CRISPR-Cas Systems ; Dysbiosis/drug therapy ; Humans
Chemical Substances	Anti-Bacterial Agents
Language	English
Publishing date	2021-09-11
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Review
ZDB-ID	645085-4
ISSN	1473-6527 ; 1535-3877 ; 0951-7375 ; 1355-834X
ISSN (online)	1473-6527 ; 1535-3877
ISSN	0951-7375 ; 1355-834X
DOI	10.1097/QCO.0000000000000772
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Zs.A 2462: Show issues

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Je nach Verfügbarkeit (siehe Angabe bei Bestand)
bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular
Jg. 1995 - 2021: Lesesall (2.OG)
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Article ; Online: A foundational approach to culture and analyze malnourished organoids.

Perlman, Meryl / Senger, Stefania / Verma, Smriti / Carey, James / Faherty, Christina S

Gut microbes

2023 Volume 15, Issue 2, Page(s) 2248713

Abstract: The gastrointestinal (GI) epithelium plays a major role in nutrient absorption, barrier formation, and innate immunity. The development of organoid-based methodology has significantly impacted the study of the GI epithelium, particularly in the fields of ...

Abstract	The gastrointestinal (GI) epithelium plays a major role in nutrient absorption, barrier formation, and innate immunity. The development of organoid-based methodology has significantly impacted the study of the GI epithelium, particularly in the fields of mucosal biology, immunity, and host-microbe interactions. Various effects on the GI epithelium, such as genetics and nutrition, impact patients and alter disease states. Thus, incorporating these effects into organoid-based models will facilitate a better understanding of disease progression and offer opportunities to evaluate therapeutic candidates. One condition that has a significant effect on the GI epithelium is malnutrition, and studying the mechanistic impacts of malnutrition would enhance our understanding of several pathologies. Therefore, the goal of this study was to begin to develop methodology to generate viable malnourished organoids with accessible techniques and resources that can be used for a wide array of mechanistic studies. By selectively limiting distinct macronutrient components of organoid media, we were able to successfully culture and evaluate malnourished organoids. Genetic and protein-based analyses were used to validate the approach and confirm the presence of known biomarkers of malnutrition. Additionally, as proof-of-concept, we utilized malnourished organoid-derived monolayers to evaluate the effect of malnourishment on barrier formation and the ability of the bacterial pathogen
MeSH term(s)	Humans ; Gastrointestinal Microbiome ; Malnutrition ; Nutritional Status ; Epithelium ; Organoids
Language	English
Publishing date	2023-09-19
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2575755-6
ISSN	1949-0984 ; 1949-0984
ISSN (online)	1949-0984
ISSN	1949-0984
DOI	10.1080/19490976.2023.2248713
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Bile Salt-induced Biofilm Formation in Enteric Pathogens: Techniques for Identification and Quantification.

Nickerson, Kourtney P / Faherty, Christina S

Journal of visualized experiments : JoVE

2018 , Issue 135

Abstract: Biofilm formation is a dynamic, multistage process that occurs in bacteria under harsh environmental conditions or times of stress. For enteric pathogens, a significant stress response is induced during gastrointestinal transit and upon bile exposure, a ... ...

Abstract	Biofilm formation is a dynamic, multistage process that occurs in bacteria under harsh environmental conditions or times of stress. For enteric pathogens, a significant stress response is induced during gastrointestinal transit and upon bile exposure, a normal component of human digestion. To overcome the bactericidal effects of bile, many enteric pathogens form a biofilm hypothesized to permit survival when transiting through the small intestine. Here we present methodologies to define biofilm formation through solid-phase adherence assays as well as extracellular polymeric substance (EPS) matrix detection and visualization. Furthermore, biofilm dispersion assessment is presented to mimic the analysis of events triggering release of bacteria during the infection process. Crystal violet staining is used to detect adherent bacteria in a high-throughput 96-well plate adherence assay. EPS production assessment is determined by two assays, namely microscopy staining of the EPS matrix and semi-quantitative analysis with a fluorescently-conjugated polysaccharide binding lectin. Finally, biofilm dispersion is measured through colony counts and plating. Positive data from multiple assays support the characterization of biofilms and can be utilized to identify bile salt-induced biofilm formation in other bacterial strains.
MeSH term(s)	Anti-Bacterial Agents/pharmacology ; Anti-Bacterial Agents/therapeutic use ; Bacteria/pathogenicity ; Bile Acids and Salts/metabolism ; Biofilms/growth & development ; Humans
Chemical Substances	Anti-Bacterial Agents ; Bile Acids and Salts
Language	English
Publishing date	2018-05-06
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Video-Audio Media
ISSN	1940-087X
ISSN (online)	1940-087X
DOI	10.3791/57322
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: Spheres of Influence: Insights into

Verma, Smriti / Senger, Stefania / Cherayil, Bobby J / Faherty, Christina S

Microorganisms

2020 Volume 8, Issue 4

Abstract: The molecular complexity of host-pathogen interactions remains poorly understood in many infectious diseases, particularly in humans due to the limited availability of reliable and specific experimental models. To bridge the gap between classical two- ... ...

Abstract	The molecular complexity of host-pathogen interactions remains poorly understood in many infectious diseases, particularly in humans due to the limited availability of reliable and specific experimental models. To bridge the gap between classical two-dimensional culture systems, which often involve transformed cell lines that may not have all the physiologic properties of primary cells, and in vivo animal studies, researchers have developed the organoid model system. Organoids are complex three-dimensional structures that are generated in vitro from primary cells and can recapitulate key in vivo properties of an organ such as structural organization, multicellularity, and function. In this review, we discuss how organoids have been deployed in exploring
Language	English
Publishing date	2020-04-01
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	2720891-6
ISSN	2076-2607
ISSN	2076-2607
DOI	10.3390/microorganisms8040504
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Article ; Online: Spheres of Influence

Smriti Verma / Stefania Senger / Bobby J. Cherayil / Christina S. Faherty

Microorganisms, Vol 8, Iss 504, p

Insights into Salmonella Pathogenesis from Intestinal Organoids

2020 Volume 504

Abstract	The molecular complexity of host-pathogen interactions remains poorly understood in many infectious diseases, particularly in humans due to the limited availability of reliable and specific experimental models. To bridge the gap between classical two-dimensional culture systems, which often involve transformed cell lines that may not have all the physiologic properties of primary cells, and in vivo animal studies, researchers have developed the organoid model system. Organoids are complex three-dimensional structures that are generated in vitro from primary cells and can recapitulate key in vivo properties of an organ such as structural organization, multicellularity, and function. In this review, we discuss how organoids have been deployed in exploring Salmonella infection in mice and humans. In addition, we summarize the recent advancements that hold promise to elevate our understanding of the interactions and crosstalk between multiple cell types and the microbiota with Salmonella . These models have the potential for improving clinical outcomes and future prophylactic and therapeutic intervention strategies.
Keywords	organoids ; enteroids ; Salmonella ; host-pathogen interactions ; model systems ; infectious diseases ; Biology (General) ; QH301-705.5
Subject code	612
Language	English
Publishing date	2020-04-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Novel Bacteroides Vulgatus strain protects against gluten-induced break of human celiac gut epithelial homeostasis: a pre-clinical proof-of-concept study.

Tran, Tina / Senger, Stefania / Baldassarre, Mariella / Brosnan, Rachel A / Cristofori, Fernanda / Crocco, Marco / De Santis, Stefania / Elli, Luca / Faherty, Christina S / Francavilla, Ruggero / Goodchild-Michelman, Isabella / Kenyon, Victoria A / Leonard, Maureen M / Lima, Rosiane S / Malerba, Federica / Montuori, Monica / Morelli, Annalisa / Norsa, Lorenzo / Passaro, Tiziana /

Piemontese, Pasqua / Reed, James C / Sansotta, Naire / Valitutti, Francesco / Zomorrodi, Ali R / Fasano, Alessio

Pediatric research

2024 Volume 95, Issue 5, Page(s) 1254–1264

Abstract: Background and aims: We have identified a decreased abundance of microbial species known to have a potential anti-inflammatory, protective effect in subjects that developed Celiac Disease (CeD) compared to those who did not. We aim to confirm the ... ...

Abstract	Background and aims: We have identified a decreased abundance of microbial species known to have a potential anti-inflammatory, protective effect in subjects that developed Celiac Disease (CeD) compared to those who did not. We aim to confirm the potential protective role of one of these species, namely Bacteroides vulgatus, and to mechanistically establish the effect of bacterial bioproducts on gluten-dependent changes on human gut epithelial functions. Methods: We identified, isolated, cultivated, and sequenced a unique novel strain (20220303-A2) of B. vulgatus found only in control subjects. Using a human gut organoid system developed from pre-celiac patients, we monitored epithelial phenotype and innate immune cytokines at baseline, after exposure to gliadin, or gliadin plus B. vulgatus cell free supernatant (CFS). Results: Following gliadin exposure, we observed increases in epithelial cell death, epithelial monolayer permeability, and secretion of pro-inflammatory cytokines. These effects were mitigated upon exposure to B. vulgatus 20220303-A2 CFS, which had matched phenotype gene product mutations. These protective effects were mediated by epigenetic reprogramming of the organoids treated with B. vulgatus CFS. Conclusions: We identified a unique strain of B. vulgatus that may exert a beneficial role by protecting CeD epithelium against a gluten-induced break of epithelial tolerance through miRNA reprogramming. Impact: Gut dysbiosis precedes the onset of celiac disease in genetically at-risk infants. This dysbiosis is characterized by the loss of protective bacterial strains in those children who will go on to develop celiac disease. The paper reports the mechanism by which one of these protective strains, B. vulgatus, ameliorates the gluten-induced break of gut epithelial homeostasis by epigenetically re-programming the target intestinal epithelium involving pathways controlling permeability, immune response, and cell turnover.
Language	English
Publishing date	2024-01-04
Publishing country	United States
Document type	Journal Article
ZDB-ID	4411-8
ISSN	1530-0447 ; 0031-3998
ISSN (online)	1530-0447
ISSN	0031-3998
DOI	10.1038/s41390-023-02960-0
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Zs.A 564: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (1.OG) ab Jg. 2022: Lesesaal (EG)
Zs.MO 373: Show issues

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Article: Spheres of Influence: Insights into Salmonella Pathogenesis from Intestinal Organoids

Verma, Smriti / Senger, Stefania / Cherayil, Bobby J. / Faherty, Christina S.

Microorganisms. 2020 Apr. 01, v. 8, no. 4

2020

Abstract	The molecular complexity of host-pathogen interactions remains poorly understood in many infectious diseases, particularly in humans due to the limited availability of reliable and specific experimental models. To bridge the gap between classical two-dimensional culture systems, which often involve transformed cell lines that may not have all the physiologic properties of primary cells, and in vivo animal studies, researchers have developed the organoid model system. Organoids are complex three-dimensional structures that are generated in vitro from primary cells and can recapitulate key in vivo properties of an organ such as structural organization, multicellularity, and function. In this review, we discuss how organoids have been deployed in exploring Salmonella infection in mice and humans. In addition, we summarize the recent advancements that hold promise to elevate our understanding of the interactions and crosstalk between multiple cell types and the microbiota with Salmonella. These models have the potential for improving clinical outcomes and future prophylactic and therapeutic intervention strategies.
Keywords	Salmonella ; intestines ; microorganisms ; organoids ; pathogenesis ; salmonellosis ; therapeutics
Language	English
Dates of publication	2020-0401
Publishing place	Multidisciplinary Digital Publishing Institute
Document type	Article
ZDB-ID	2720891-6
ISSN	2076-2607
ISSN	2076-2607
DOI	10.3390/microorganisms8040504
Database	NAL-Catalogue (AGRICOLA)

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Article: Bile salt-induced biofilm formation in enteric pathogens: techniques for identification and quantification

Nickerson, Kourtney P / Faherty, Christina S

Journal of visualized experiments. 2018 May 06, , no. 135

2018

Abstract	Biofilm formation is a dynamic, multistage process that occurs in bacteria under harsh environmental conditions or times of stress. For enteric pathogens, a significant stress response is induced during gastrointestinal transit and upon bile exposure, a normal component of human digestion. To overcome the bactericidal effects of bile, many enteric pathogens form a biofilm hypothesized to permit survival when transiting through the small intestine. Here we present methodologies to define biofilm formation through solid-phase adherence assays as well as extracellular polymeric substance (EPS) matrix detection and visualization. Furthermore, biofilm dispersion assessment is presented to mimic the analysis of events triggering release of bacteria during the infection process. Crystal violet staining is used to detect adherent bacteria in a high-throughput 96-well plate adherence assay. EPS production assessment is determined by two assays, namely microscopy staining of the EPS matrix and semi-quantitative analysis with a fluorescently-conjugated polysaccharide binding lectin. Finally, biofilm dispersion is measured through colony counts and plating. Positive data from multiple assays support the characterization of biofilms and can be utilized to identify bile salt-induced biofilm formation in other bacterial strains.
Keywords	antibacterial properties ; bacteria ; bile ; biofilm ; digestion ; enteropathogens ; environmental factors ; gastrointestinal transit ; gentian violet ; humans ; lectins ; microscopy ; polymers ; polysaccharides ; small intestine ; staining ; stress response
Language	English
Dates of publication	2018-0506
Size	p. e57322.
Publishing place	Journal of Visualized Experiments
Document type	Article
ZDB-ID	2259946-0
ISSN	1940-087X
ISSN	1940-087X
DOI	10.3791/57322
Database	NAL-Catalogue (AGRICOLA)

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