Name: Ontwikkeling en beoordeling van intracellulaire infectiemodellen voor Staphylococcus aureus (Video) | JoVE Journal
Uploaded: 2025-01-17T14:50:00.000+00:00
Duration: 8 min 32 s
Description: Leer een intracellulair infectiemodel voor Staphylococcus aureus ontwikkelen met celkweek en gentamicine-behandeling. Deze methode helpt onderzoekers overleving van bacteriën in macrofagen bestuderen.

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<meta charset="utf8"></head><body>Geoptimaliseerde in-vivomodellen voor intracellulaire S. aureus-infectiestudies

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<li>Wittekind, M. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+novel+protein+scra+acts+through+the+saers+two-component+system+to+regulate+virulence+gene+expression+in+Staphylococcus+aureus%5BTitle%5D)+AND+%22Mol+Microbiol%22%5BJournal%5D)">The novel protein scra acts through the saers two-component system to regulate virulence gene expression in Staphylococcus aureus.</a> Mol Microbiol. 117 (5), 1196-1212 (2022).</li>
<li>Pidwill, G. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+role+of+macrophages+in+Staphylococcus+aureus+infection%5BTitle%5D)+AND+%22Front+Immunol%22%5BJournal%5D)">The role of macrophages in Staphylococcus aureus infection.</a> Front Immunol. 11, 620339(2020).</li>
<li>Lang, J. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+photoconvertible+reporter+system+for+bacterial+metabolic+activity+reveals+that+Staphylococcus+aureus+enters+a+dormant-like+state+to+persist+within+macrophages%5BTitle%5D)+AND+%22mBio%22%5BJournal%5D)">A photoconvertible reporter system for bacterial metabolic activity reveals that Staphylococcus aureus enters a dormant-like state to persist within macrophages.</a> mBio. 13 (5), e0231622(2022).</li>
<li>Li, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Interactions+between+macrophages+and+biofilm+during+Staphylococcus+aureus-associated+implant+infection%3A+Difficulties+and+solutions%5BTitle%5D)+AND+%22J+Innate+Immun%22%5BJournal%5D)">Interactions between macrophages and biofilm during Staphylococcus aureus-associated implant infection: Difficulties and solutions.</a> J Innate Immun. 15 (1), 499-515 (2023).</li>
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<li>Mulcahy, M. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Manipulation+of+autophagy+and+apoptosis+facilitates+intracellular+survival+of+Staphylococcus+aureus+in+human+neutrophils%5BTitle%5D)+AND+%22Front+Immunol%22%5BJournal%5D)">Manipulation of autophagy and apoptosis facilitates intracellular survival of Staphylococcus aureus in human neutrophils.</a> Front Immunol. 11, 565545(2020).</li>
<li>Cai, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Staphylococcus+aureus+facilitates+its+survival+in+bovine+macrophages+by+blocking+autophagic+flux%5BTitle%5D)+AND+%22J+Cell+Mol+Med%22%5BJournal%5D)">Staphylococcus aureus facilitates its survival in bovine macrophages by blocking autophagic flux.</a> J Cell Mol Med. 24 (6), 3460-3468 (2020).</li>
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<li>Kelly, J. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Measurement+of+accumulation+of+antibiotics+to+Staphylococcus+aureus+in+phagosomes+of+live+macrophages%5BTitle%5D)+AND+%22Angew+Chem+Int+Ed+Engl%22%5BJournal%5D)">Measurement of accumulation of antibiotics to Staphylococcus aureus in phagosomes of live macrophages.</a> Angew Chem Int Ed Engl. 63 (3), e202313870(2024).</li>
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<li>Lehar, S. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Novel+antibody-antibiotic+conjugate+eliminates+intracellular+S.+aureus%5BTitle%5D)+AND+%22Nature%22%5BJournal%5D)">Novel antibody-antibiotic conjugate eliminates intracellular S. aureus.</a> Nature. 527 (7578), 323-328 (2015).</li>
<li>Peyrusson, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Intracellular+Staphylococcus+aureus+persisters+upon+antibiotic+exposure%5BTitle%5D)+AND+%22Nat+Commun%22%5BJournal%5D)">Intracellular Staphylococcus aureus persisters upon antibiotic exposure.</a> Nat Commun. 11 (1), 2200(2020).</li>
<li>Pollitt, E. J. G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Staphylococcus+aureus+infection+dynamics%5BTitle%5D)+AND+%22PLoS+Pathog%22%5BJournal%5D)">Staphylococcus aureus infection dynamics.</a> PLoS Pathog. 14 (6), e1007112(2018).</li>
<li>Sanders, E. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Aseptic+laboratory+techniques%3A+plating+methods%5BTitle%5D)+AND+%22J+Vis+Exp%22%5BJournal%5D)">Aseptic laboratory techniques: plating methods.</a> J Vis Exp. 11 (63), e3064(2012).</li>
<li>Qin, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Antibody-antibiotic+conjugate+targeted+therapy+for+orthopedic+implant-associated+intracellular+S.+aureus+infections%5BTitle%5D)+AND+%22J+Adv+Res%22%5BJournal%5D)">Antibody-antibiotic conjugate targeted therapy for orthopedic implant-associated intracellular S. aureus infections.</a> J Adv Res. 65, 239-255 (2024).</li>
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</ol>

<table><tbody><tr><td>24-well plate</td><td>Corning Incorporated, USA</td><td>3524</td><td></td></tr><tr><td>4 % paraformaldehyde solutione</td><td>BBI, UK</td><td>E672002-0500</td><td></td></tr><tr><td>6-well plate</td><td>Corning Incorporated, USA</td><td>3516</td><td></td></tr><tr><td>Beef extract powder</td><td>BBI, UK</td><td>A600114-0500</td><td></td></tr><tr><td>Biohazard safety equipment</td><td>Heal force, China</td><td>VS-1300L-u</td><td></td></tr><tr><td>Cell incubator</td><td>ESCO, Singapore</td><td>CCL-170B-8</td><td></td></tr><tr><td>Cell scraper</td><td>Nest</td><td>710001</td><td></td></tr><tr><td>Centrifuge M1416R</td><td>RWD, China</td><td>M1416R</td><td></td></tr><tr><td>Centrifuge tube</td><td>Guanghou Labselect, China</td><td>CT-002-50A</td><td></td></tr><tr><td>Confocal laser scanning microscope (CLSM)</td><td>Zeiss, Germany</td><td>880</td><td></td></tr><tr><td>Confocal petri dish</td><td>Biosharp, China</td><td>BS-20-GJM</td><td></td></tr><tr><td>DAPI dye</td><td>Shanghai Beyotime, China&nbsp;</td><td>C1006</td><td></td></tr><tr><td>DIL working fluid</td><td>Shanghai Beyotime, China&nbsp;</td><td>C1991S</td><td></td></tr><tr><td>Dulbecco&rsquo;s Modified Eagle Medium</td><td>Thermo Gibco, USA</td><td>C11995500BT</td><td></td></tr><tr><td>Fetal Bovine Serum</td><td>Hyclone</td><td>SV30208.02</td><td></td></tr><tr><td>Gentamycin</td><td>Shanghai Sangon, China</td><td>B540724-0010</td><td></td></tr><tr><td>Incubator</td><td>Shanghai Hengzi, China</td><td>HDPF-150</td><td></td></tr><tr><td>Lysozyme</td><td>Beijing Solarbio, China</td><td>L9070</td><td></td></tr><tr><td>MRSA252</td><td>Third Military Medical University, China</td><td>null</td><td></td></tr><tr><td>MRSA252(GPF)</td><td>Third Military Medical University, China</td><td>null</td><td></td></tr><tr><td>Penicillin and Streptomycin</td><td>Shanghai Beyotime, China&nbsp;</td><td>C0222</td><td></td></tr><tr><td>Phosphate Buffer Solution</td><td>Shanghai Beyotime, China&nbsp;</td><td>ST476</td><td></td></tr><tr><td>Saline</td><td>Sichuan Kelun, China</td><td>null</td><td></td></tr><tr><td>Sodium chloride</td><td>Shanghai Macklin, China</td><td>S805275</td><td></td></tr><tr><td>Starch soluble</td><td>Shanghai Sangon, China</td><td>A500904-0500</td><td></td></tr><tr><td>Triton X-100</td><td>Shanghai Beyotime, China&nbsp;</td><td>P0096-100ml</td><td></td></tr><tr><td>Tryptic Soy Agar (TSA) plates</td><td>Beijing AOBOX Biotechnology Co., LTD,China</td><td>02-130</td><td></td></tr><tr><td>Tryptic Soy Broth (TSB) medium</td><td>Beijing AOBOX Biotechnology Co., LTD,China</td><td>02-102K</td><td></td></tr><tr><td>Tryptone</td><td>OXOID, UK</td><td>LP0042B</td><td></td></tr><tr><td>Vancomycin</td><td>Shanghai Beyotime, China&nbsp;</td><td>ST2807-250mg</td><td></td></tr><tr><td>RAW264.7 cell</td><td>USA, ATCC</td><td>null</td><td></td></tr></tbody></table>

ontwikkeling en beoordeling van intracellulaire infectiemodellen voor staphylococcus aureus

Development and Assessment of Intracellular Infection Models for Staphylococcus aureus

Ontwikkeling en beoordeling van intracellulaire infectiemodellen voor Staphylococcus aureus

S. aureus can invade and persist within host cells, including immune cells, which allows it to evade immune detection and clearance. This intracellular ...

development-assessment-intracellular-infection-models-for

Research

JoVE Journal

Immunology and Infection

Immunologie en Infectie

Een muismodel voor huid- en weke deleninfecties wordt gebruikt om de virulentiefunctie van methicilline-resistente Staphylococcus aureus (MRSA) en de immunologische respons van de gastheer te beoordelen. Hier presenteren wij een subcutaan infectiemodel voor huid- en weke deleninfecties.

Subcutane Infectie van Methicilline Resistente Staphylococcus Aureus (MRSA)

Experimentele rat endocarditis model veroorzaakt door...

Experimentele Endocarditis Model van Methicilline Resistente<em&gt; Staphylococcus aureus</em&gt; (MRSA) in Rat

Source: Zhang, C. et al. Development and Assessment of Intracellular Infection Models for Staphylococcus aureus. J. Vis. Exp. (2025)This video demonstrates the quantification of intracellular survival of a methicillin-resistant bacterial pathogen in mouse peritoneal macrophages. It outlines the steps involved in infection, extracellular bacterial clearance, host cell lysis, and colony enumeration to assess bacterial survival within host macrophages.

Assessing the Survival of a Bacterial Pathogen in Mouse Peritoneal Macrophages

Source: Zhang, C., et al., Development and Assessment of Intracellular Infection Models for Staphylococcus aureus. J. Vis. Exp. (2025)The video demonstrates the isolation of kidneys containing Staphylococcus aureus-infected macrophages, followed by tissue homogenization to release intracellular bacteria. It quantifies bacterial persistence within macrophages in control and antibiotic-treated groups by counting colony-forming units.

Assessing Intracellular Bacterial Persistence and Antibiotic Resistance in a Mouse Model

De huidige studie beschrijft een zebravis embryo model voor in vivo visualisatie en intravital analyse van biomaterial-geassocieerde infectie na verloop van tijd op basis van fluorescentie microscopie. Dit model is een veelbelovende aanvulling op zoogdieren dierlijke modellen zoals Muismodellen voor het bestuderen van de biomaterial-geassocieerde infecties in vivo.

Een zebravis Embryo Model voor In Vivo visualisatie en Intravital analyse van Biomaterial-geassocieerde goudhoudende Stafylokok infectie

Een aanpak wordt beschreven voor real-time detectie van de ingeboren immune reactie op het verwonden van cutane en Staphylococcus aureus -infectie van muizen. Door vergelijking LysM-EGFP muizen (die bezitten fluorescerende neutrofielen) met een LysM-EGFP gekruist immunodeficiëntie muis stam, we vooraf ons begrip van de infectie en de ontwikkeling van benaderingen ter bestrijding van de infectie.

Een muismodel te beoordelen ingeboren immuunreactie op Staphylococcus aureus -infectie

Synthetisch cystic fibrosis sputum medium (SCFM2) kan worden gebruikt in combinatie met zowel confocale laserscanmicroscopie als fluorescentie-geactiveerde celsortering om bacteriële aggregaten met hoge resolutie te observeren. Dit artikel beschrijft methoden om geaggregeerde populaties te beoordelen tijdens antimicrobiële behandeling als een platform voor toekomstige studies.

Tools voor de real-time beoordeling van een Pseudomonas aeruginosa-infectiemodel

Dit protocol heeft tot doel te beschrijven hoe de mate van Staphylococcus aureus internalisatie en het vermogen om te overleven in de menselijke gastheercel kan worden bestudeerd, evenals de intracellulaire werkzaamheid van antimicrobiële verbindingen.

Verbeterde enzymbeschermingstest om staphylococcus aureus internalisatie en intracellulaire werkzaamheid van antimicrobiële verbindingen te bestuderen

Dit protocol beschrijft de studie van neutrofiel-biofilm interacties. Staphylococcus aureus biofilms worden in vitro vastgesteld en geïncubeerd met perifere bloed-afgeleide menselijke neutrofielen. De oxidatieve burst-respons van neutrofielen wordt gekwantificeerd en de neutrofiellokalisatie in de biofilm wordt bepaald door microscopie.

Gestandaardiseerde in vitro assays om interacties tussen menselijke neutrofielen en staphylococcus aureus biofilms te visualiseren en te kwantificeren

Het protocol beschrijft een stapsgewijze methode om een ex vivo schapen gewonde huidmodel op te zetten dat geïnfecteerd is met Staphylococcus aureus. Dit high-throughput model simuleert infecties beter in vivo in vergelijking met conventionele microbiologische technieken en biedt onderzoekers een fysiologisch relevant platform om de werkzaamheid van opkomende antimicrobiële stoffen te testen.

Een nieuw ex vivo schapenhuidwondmodel met hoge doorvoer voor het testen van opkomende antibiotica

Het huidige protocol beschrijft een muismodel dat gebruik maakt van subcutane dorsale implantatie om implantaat-geassocieerde infecties te onderzoeken, waardoor uitgebreid onderzoek naar pathofysiologische mechanismen mogelijk wordt en de ontwikkeling van diagnostische criteria met gerichte therapeutische strategieën wordt ondersteund.

Diermodel van implantaat-geassocieerde infecties bij muizen

Leer een intracellulair infectiemodel voor Staphylococcus aureus ontwikkelen met celkweek en gentamicine-behandeling. Deze methode helpt onderzoekers overleving van bacteriën in macrofagen bestuderen.

Ontwikkeling en beoordeling van intracellulaire infectiemodellen voor Staphylococcus aureus

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