Standardized In vitro Assays to Visualize and Quantify Interactions between Human Neutrophils and Staphylococcus aureus Biofilms

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dc.contributor.authorRana, Pranav S. J. B.en
dc.contributor.authorGloag, Erin S.en
dc.contributor.authorWozniak, Daniel J.en
dc.date.accessioned2022-12-14T20:15:47Zen
dc.date.available2022-12-14T20:15:47Zen
dc.date.issued2022-06-08en
dc.date.updated2022-12-14T19:06:52Zen
dc.description.abstractNeutrophils are the first line of defense deployed by the immune system during microbial infection. In vivo, neutrophils are recruited to the site of infection where they use processes such as phagocytosis, production of reactive oxygen and nitrogen species (ROS, RNS, respectively), NETosis (neutrophil extracellular trap), and degranulation to kill microbes and resolve the infection. Interactions between neutrophils and planktonic microbes have been extensively studied. There have been emerging interests in studying infections caused by biofilms in recent years. Biofilms exhibit properties, including tolerance to killing by neutrophils, distinct from their planktonic-grown counterparts. With the successful establishment of both in vitro and in vivo biofilm models, interactions between these microbial communities with different immune cells can now be investigated. Here, techniques that use a combination of traditional biofilm models and well-established neutrophil activity assays are tailored specifically to study neutrophil and biofilm interactions. Wide-field fluorescence microscopy is used to monitor the localization of neutrophils in biofilms. These biofilms are grown in static conditions, followed by the addition of neutrophils derived from human peripheral blood. The samples are stained with appropriate dyes prior to visualization under the microscope. Additionally, the production of ROS, which is one of the many neutrophil responses against pathogens, is quantified in the presence of a biofilm. The addition of immune cells to this established system will expand the understanding of host-pathogen interactions while ensuring the use of standardized and optimized conditions to measure these processes accurately.en
dc.description.versionPublished versionen
dc.format.mimetypeapplication/pdfen
dc.identifier.doihttps://doi.org/10.3791/63773en
dc.identifier.eissn1940-087Xen
dc.identifier.issn1940-087Xen
dc.identifier.issue184en
dc.identifier.orcidGloag, Erin [0000-0001-8895-3444]en
dc.identifier.pmid35758715en
dc.identifier.urihttp://hdl.handle.net/10919/112888en
dc.language.isoenen
dc.publisherMyJove Corporationen
dc.relation.urihttps://www.ncbi.nlm.nih.gov/pubmed/35758715en
dc.rightsCreative Commons Attribution-NonCommercial-NoDerivatives 4.0 Internationalen
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/en
dc.subjectEmerging infectious diseasesen
dc.subjectBiodefenseen
dc.subjectVaccine relateden
dc.subjectPreventionen
dc.subjectInfectious diseasesen
dc.subject5 Development of treatments and therapeutic interventionsen
dc.subject5.1 Pharmaceuticalsen
dc.subject2.2 Factors relating to the physical environmenten
dc.subject2 Aetiologyen
dc.subjectInfectionen
dc.subject.meshBiofilmsen
dc.subject.meshExtracellular Trapsen
dc.subject.meshHumansen
dc.subject.meshNeutrophilsen
dc.subject.meshPlanktonen
dc.subject.meshReactive Oxygen Speciesen
dc.subject.meshStaphylococcal Infectionsen
dc.subject.meshStaphylococcus aureusen
dc.titleStandardized <i>In vitro</i> Assays to Visualize and Quantify Interactions between Human Neutrophils and <i>Staphylococcus aureus</i> Biofilmsen
dc.title.serialJournal of Visualized Experimentsen
dc.typeArticle - Refereeden
dc.type.dcmitypeTexten
dc.type.otherJournal Articleen
dc.type.otherResearch Support, N.I.H., Extramuralen
dc.type.otherResearch Support, Non-U.S. Gov'ten
pubs.organisational-group/Virginia Techen
pubs.organisational-group/Virginia Tech/Veterinary Medicineen
pubs.organisational-group/Virginia Tech/Veterinary Medicine/Biomedical Sciences and Pathobiologyen
pubs.organisational-group/Virginia Tech/All T&R Facultyen
pubs.organisational-group/Virginia Tech/Veterinary Medicine/CVM T&R Facultyen

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