Analysis of Epithelial Cell Responses to Microbial Pathogens

Spyridoula Nikou; Jemima Ho; Olivia Hepworth; Nicole Ponde; Ruth Dickenson; Jonathan P. Richardson; Julian R. Naglik

doi:10.1007/978-1-0716-1182-1_5

Analysis of Epithelial Cell Responses to Microbial Pathogens

Spyridoula Nikou, Jemima Ho, Olivia Hepworth, Nicole Ponde, Ruth Dickenson, Jonathan P. Richardson, Julian R. Naglik

King's College London

Research output: Contribution to journal › Article › peer-review

Abstract

The epithelial cell is usually the first host cell that interacts with the microbiota present at mucosal surfaces. Although initially thought of as "bystander" cells with barrier function, the epithelial cell is now known to be a sentinel cell in the recognition and discrimination of commensal and pathogenic microorganisms and a key cell in initiating subsequent innate and adaptive immune responses. Here, we describe the main assays utilized in analyzing the activation of epithelial cell signaling (western blotting), transcription factors (TransAm), gene expression (quantitative reverse transcription PCR (qRT-PCR)), cytokine responses (ELISA, Luminex), and damage induction (lactate dehydrogenase (LDH) release). While our laboratory focuses on the epithelial response to Candida pathogens, these assays can be applied universally to analyze the activation of epithelial cells in response to any microbial pathogen.

Original language	English
Pages (from-to)	49-82
Number of pages	34
Journal	Methods in molecular biology (Clifton, N.J.)
Volume	2260
DOIs	https://doi.org/10.1007/978-1-0716-1182-1_5
Publication status	Published - 2021

Keywords

Cell damage
Cell signaling
Cytokine
ELISA
Gene expression
Lactate dehydrogenase
Luminex
qRT-PCR
TransAm
Transcription factor
Western blotting

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abstract = "The epithelial cell is usually the first host cell that interacts with the microbiota present at mucosal surfaces. Although initially thought of as {"}bystander{"} cells with barrier function, the epithelial cell is now known to be a sentinel cell in the recognition and discrimination of commensal and pathogenic microorganisms and a key cell in initiating subsequent innate and adaptive immune responses. Here, we describe the main assays utilized in analyzing the activation of epithelial cell signaling (western blotting), transcription factors (TransAm), gene expression (quantitative reverse transcription PCR (qRT-PCR)), cytokine responses (ELISA, Luminex), and damage induction (lactate dehydrogenase (LDH) release). While our laboratory focuses on the epithelial response to Candida pathogens, these assays can be applied universally to analyze the activation of epithelial cells in response to any microbial pathogen.",

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note = "Funding Information: This work was supported by grants from the Medical Research Council (MR/M011372/1), Biotechnology and Biological Sciences Research Council (BB/N014677/1), National Institutes of Health (R37-DE022550), King?s Health Partners Challenge Fund (R170501), the Rosetrees Trust (M680), and the NIH Research at Guys and St. Thomas?s NHS Foundation Trust and the King?s College London Biomedical Research Centre (IS-BRC-1215-20006). Publisher Copyright: {\textcopyright} 2021, Springer Science+Business Media, LLC, part of Springer Nature. Copyright: Copyright 2021 Elsevier B.V., All rights reserved.",

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doi = "10.1007/978-1-0716-1182-1_5",

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AU - Ponde, Nicole

AU - Dickenson, Ruth

AU - Richardson, Jonathan P.

AU - Naglik, Julian R.

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Y1 - 2021

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Analysis of Epithelial Cell Responses to Microbial Pathogens

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Keywords

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