Name: Desenvolvimento e Avaliação de Modelos de Infecção Intracelular para Staphylococcus aureus (Vídeo) | JoVE Journal
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Description: Aprenda a desenvolver modelos de infecção intracelular para Staphylococcus aureus usando cultura celular e sequenciamento de alta vazão. Este método ajuda pesquisadores a estudar mecanismos bacterianos e otimizar estratégias terapêuticas.

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<meta charset="utf8"></head><body>Modelos in vivo otimizados para estudos de infecção intracelular por S. aureus

<ol><li>Sutton, J. A. F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Staphylococcus+aureus+cell+wall+structure+and+dynamics+during+host-pathogen+interaction%5BTitle%5D)+AND+%22PLoS+Pathog%22%5BJournal%5D)">Staphylococcus aureus cell wall structure and dynamics during host-pathogen interaction.</a> PLoS Pathog. 17 (3), e1009468(2021).</li>
<li>Yang, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Lateral+flow+assay+of+methicillin-resistant+Staphylococcus+aureus+using+bacteriophage+cellular+wall-binding+domain+as+recognition+agent%5BTitle%5D)+AND+%22Biosens+Bioelectron%22%5BJournal%5D)">Lateral flow assay of methicillin-resistant Staphylococcus aureus using bacteriophage cellular wall-binding domain as recognition agent.</a> Biosens Bioelectron. 182, 113189(2021).</li>
<li>Howden, B. P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Staphylococcus+aureus+host+interactions+and+adaptation%5BTitle%5D)+AND+%22Nat+Rev+Microbiol%22%5BJournal%5D)">Staphylococcus aureus host interactions and adaptation.</a> Nat Rev Microbiol. 21 (6), 380-395 (2023).</li>
<li>Guo, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Biofilm+and+small+colony+variants-an+update+on+Staphylococcus+aureus+strategies+toward+drug+resistance%5BTitle%5D)+AND+%22Int+J+Mol+Sci%22%5BJournal%5D)">Biofilm and small colony variants-an update on Staphylococcus aureus strategies toward drug resistance.</a> Int J Mol Sci. 23 (3), 1241(2022).</li>
<li>Huitema, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Intracellular+escape+strategies+of+Staphylococcus+aureus+in+persistent+cutaneous+infections%5BTitle%5D)+AND+%22Exp+Dermatol%22%5BJournal%5D)">Intracellular escape strategies of Staphylococcus aureus in persistent cutaneous infections.</a> Exp Dermatol. 30 (10), 1428-1439 (2021).</li>
<li>Pidwill, G. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Clonal+population+expansion+of+Staphylococcus+aureus+occurs+due+to+escape+from+a+finite+number+of+intraphagocyte+niches%5BTitle%5D)+AND+%22Sci+Rep%22%5BJournal%5D)">Clonal population expansion of Staphylococcus aureus occurs due to escape from a finite number of intraphagocyte niches.</a> Sci Rep. 13 (1), 1188(2023).</li>
<li>Wang, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Autophagy+in+Staphylococcus+aureus+infection%5BTitle%5D)+AND+%22Front+Cell+Infect+Microbiol%22%5BJournal%5D)">Autophagy in Staphylococcus aureus infection.</a> Front Cell Infect Microbiol. 11, 750222(2021).</li>
<li>Arya, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Identification+of+an+antivirulence+agent+targeting+the+master+regulator+of+virulence+genes+in+Staphylococcus+aureus%5BTitle%5D)+AND+%22Front+Cell+Infect+Microbiol%22%5BJournal%5D)">Identification of an antivirulence agent targeting the master regulator of virulence genes in Staphylococcus aureus.</a> Front Cell Infect Microbiol. 13, 1268044(2023).</li>
<li>Münzenmayer, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Influence+of+sae-regulated+and+agr-regulated+factors+on+the+escape+of+Staphylococcus+aureus+from+human+macrophages%5BTitle%5D)+AND+%22Cell+Microbiol%22%5BJournal%5D)">Influence of sae-regulated and agr-regulated factors on the escape of Staphylococcus aureus from human macrophages.</a> Cell Microbiol. 18 (8), 1172-1183 (2016).</li>
<li>Chin, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Staphylococcus+lugdunensis+uses+the+agr+regulatory+system+to+resist+killing+by+host+innate+immune+effectors%5BTitle%5D)+AND+%22Infect+Immun%22%5BJournal%5D)">Staphylococcus lugdunensis uses the agr regulatory system to resist killing by host innate immune effectors.</a> Infect Immun. 90 (10), e0009922(2022).</li>
<li>Podkowik, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Quorum-sensing+agr+system+of+Staphylococcus+aureus+primes+gene+expression+for+protection+from+lethal+oxidative+stress%5BTitle%5D)+AND+%22Elife%22%5BJournal%5D)">Quorum-sensing agr system of Staphylococcus aureus primes gene expression for protection from lethal oxidative stress.</a> Elife. 12, RP89098(2024).</li>
<li>Purves, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Air+pollution+induces+Staphylococcus+aureus+usa300+respiratory+tract+colonization+mediated+by+specific+bacterial+genetic+responses+involving+the+global+virulence+gene+regulators+agr+and+sae%5BTitle%5D)+AND+%22Environ+Microbiol%22%5BJournal%5D)">Air pollution induces Staphylococcus aureus usa300 respiratory tract colonization mediated by specific bacterial genetic responses involving the global virulence gene regulators agr and sae.</a> Environ Microbiol. 24 (9), 4449-4465 (2022).</li>
<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>
<li>Sun, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Staphylococcal+virulence+factor+hlgb+targets+the+endoplasmic-reticulum-resident+e3+ubiquitin+ligase+amfr+to+promote+pneumonia%5BTitle%5D)+AND+%22Nat+Microbiol%22%5BJournal%5D)">Staphylococcal virulence factor hlgb targets the endoplasmic-reticulum-resident e3 ubiquitin ligase amfr to promote pneumonia.</a> Nat Microbiol. 8 (1), 107-120 (2023).</li>
<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>
<li>Ahmad-Mansour, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Staphylococcus+aureus+toxins%3A+An+update+on+their+pathogenic+properties+and+potential+treatments%5BTitle%5D)+AND+%22Toxins%22%5BJournal%5D)">Staphylococcus aureus toxins: An update on their pathogenic properties and potential treatments.</a> Toxins. 13 (10), 677(2021).</li>
<li>Davis, J. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((How+I+manage+a+patient+with+MRSA+bacteremia%5BTitle%5D)+AND+%22Clin+Microbiol+Infect%22%5BJournal%5D)">How I manage a patient with MRSA bacteremia.</a> Clin Microbiol Infect. 28 (2), 190-194 (2022).</li>
<li>Fait, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Staphylococcus+aureus+response+and+adaptation+to+vancomycin%5BTitle%5D)+AND+%22Adv+Microb+Physiol%22%5BJournal%5D)">Staphylococcus aureus response and adaptation to vancomycin.</a> Adv Microb Physiol. 85, 201-258 (2024).</li>
<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>
<li>Rowe, S. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Recalcitrant+Staphylococcus+aureus+infections%3A+Obstacles+and+solutions%5BTitle%5D)+AND+%22Infect+Immun%22%5BJournal%5D)">Recalcitrant Staphylococcus aureus infections: Obstacles and solutions.</a> Infect Immun. 89 (4), e00694-e00720 (2021).</li>
<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>
<li>Andie, S. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Methicillin-resistant+Staphylococcus+aureus%5BTitle%5D)+AND+%22Nat+Rev+Dis+Primers%22%5BJournal%5D)">Methicillin-resistant Staphylococcus aureus.</a> Nat Rev Dis Primers. 31 (4), 18034(2018).</li>
</ol>

<table><tbody><tr><td>Placa de 24 poços</td><td>Corning Incorporated, EUA</td><td>3524</td><td></td></tr><tr><td>Solução de paraformaldeído a 4</td>% <td>BBI, Reino Unido</td><td>E672002-0500</td><td></td></tr><tr><td>Placa de 6 poços</td><td>Corning Incorporated, EUA</td><td>3516</td><td></td></tr><tr><td>Extrato de carne em pó</td><td>BBI, Reino Unido</td><td>A600114-0500</td><td></td></tr><tr><td>Equipamento de segurança de risco biológico</td><td>Força de cura, China</td><td>VS-1300L-u</td><td></td></tr><tr><td>Incubadora de células</td><td>ESCO, Cingapura</td><td>CCL-170B-8</td><td></td></tr><tr><td>Raspador de células</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>Tubo de centrífuga</td><td>Guanghou Labselect, China</td><td>CT-002-50A</td><td></td></tr><tr><td>Microscópio confocal de varredura a laser (CLSM)</td><td>Zeiss, Alemanha</td><td>880</td><td></td></tr><tr><td>Placa</td> <td>de Petri confocalBiosharp, China</td><td>BS-20-GJM</td><td></td></tr><tr><td>DAPI corante</td><td>Shanghai Beyotime, China&nbsp;</td>Fluido de trabalho <td>DIL C1006</td><td></td></tr><tr><td></td><td>Shanghai Beyotime, China&nbsp;</td><td>C1991S</td><td></td></tr><tr><td>Dulbecco' s Modified Eagle Medium</td><td>Thermo Gibco, EUA</td><td>C11995500BT</td><td></td></tr><tr><td>Hyclone</td> de soro<td>bovino fetal</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>Incubadora</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>Terceiro Militar Universidade Médica, 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>Penicilina e Estreptomicina</td><td>Shanghai Beyotime, China&nbsp;</td><td>C0222</td><td></td></tr><tr><td>Solução Tampão de Fosfato</td><td>Shanghai Beyotime, China&nbsp;</td><td>ST476</td><td></td></tr><tr><td>Salino</td><td>Sichuan Kelun, China</td><td>nulo</td><td></td></tr><tr><td>Cloreto de sódio</td><td>Shanghai Macklin, China</td><td>S805275</td><td></td></tr><tr><td>Solúvel em amido</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>Placas de ágar tripta de soja (TSA)</td><td>Pequim AOBOX Biotechnology Co., LTD, China</td><td>02-130</td><td></td></tr><tr><td>Caldo de soja tríptico (TSB) médio</td><td>Pequim AOBOX Biotechnology Co., LTD, China</td><td>02-102K</td><td></td></tr><tr><td>Triptona</td><td>OXOID, Reino Unido</td><td>LP0042B</td><td></td></tr><tr><td>Vancomicina</td><td>Xangai Beyotime, China&nbsp;</td><td>ST2807-250mg</td><td></td></tr><tr><td>RAW264.7 célula</td><td>EUA, ATCC</td><td>nulo</td><td></td></tr></tbody></table>

desenvolvimento e avaliação de modelos de infecção intracelular para staphylococcus aureus

Development and Assessment of Intracellular Infection Models for Staphylococcus aureus

Modelos de infecção intracelular de S. aureus foram estabelecidos com sucesso in vitro e in vivo. Otimizando as condições experimentais para fagocitose e estendendo a concentração e a duração do tratamento com antibióticos, alguns S. aureus sobreviveram dentro dos macrófagos (Figura 1). Para avaliar melhor a resistência a antibióticos de S. aureus, os macrófagos infectados com o MRSA252 foram tratados com antibióti...

Desenvolvimento e Avaliação de Modelos de Infecção Intracelular para 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

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JoVE Journal

Immunology and Infection

Imunologia e Infecção

Pele murina e modelo de infecção dos tecidos moles é utilizado para avaliar a função de virulência de Staphylococcus aureus resistente (MRSA) e as respostas imunológico do hospedeiro. Aqui, apresentamos um modelo de infecção subcutânea por infecção da pele e tecidos moles.

Infecção subcutânea de Staphylococcus aureus resistente (MRSA)

Endocardite modelo experimental de ratos devido à...

Modelo Experimental de Endocardite resistente à meticilina<em&gt; Staphylococcus aureus</em&gt; (MRSA) em Rato

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

O presente estudo descreve um modelo de embrião de zebrafish para visualização in vivo e intravital análise de infecção biomaterial associada ao longo do tempo com base em microscopia de fluorescência. Este modelo é um sistema promissor complementando modelos animais mamíferos, tais como modelos de rato para o estudo in vivo de infecções associadas a biomaterial.

Um modelo de embrião de Zebrafish para na visualização de Vivo e Intravital análise de biomateriais-associado Staphylococcus aureus infecção

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Um modelo do rato para avaliar a resposta imune inata à infecção de Staphylococcus aureus

O meio de escarro de fibrose cística sintética (SCFM2) pode ser utilizado em combinação com microscopia de varredura a laser confocal e classificação celular ativada por fluorescência para observar agregados bacterianos em alta resolução. Este artigo detalha métodos para avaliar populações agregadas durante o tratamento antimicrobiano como uma plataforma para estudos futuros.

Ferramentas para a avaliação em tempo real de um modelo de infecção por pseudomonas aeruginosa

Este protocolo tem como objetivo descrever como estudar a extensão da internalização do Staphylococcus aureus e sua capacidade de sobreviver dentro da célula hospedeira humana, bem como a eficácia intracelular de compostos antimicrobianos.

Ensaio de proteção de enzimas melhorada para estudar a internalização staphylococcus aureus e eficácia intracelular de compostos antimicrobianos

O presente protocolo descreve o estudo das interações neutrófilo-biofilme. Os biofilmes de Staphylococcus aureus são estabelecidos in vitro e incubados com neutrófilos humanos derivados do sangue periférico. A resposta de explosão oxidativa dos neutrófilos é quantificada e a localização dos neutrófilos dentro do biofilme é determinada por microscopia.

Ensaios In vitro padronizados para visualizar e quantificar interações entre neutrófilos humanos e biofilmes de Staphylococcus aureus

O protocolo descreve um método passo-a-passo para configurar um modelo de pele ferida ovina ex vivo infectada com Staphylococcus aureus. Este modelo de alto rendimento simula melhor infecções in vivo em comparação com técnicas convencionais de microbiologia e apresenta aos pesquisadores uma plataforma fisiologicamente relevante para testar a eficácia de antimicrobianos emergentes.

Um novo modelo de ferida cutânea de ovino ex vivo de alto rendimento para testar antibióticos emergentes

O presente protocolo descreve um modelo de camundongo que emprega implante dorsal subcutâneo para investigar infecções associadas ao implante, permitindo uma investigação abrangente dos mecanismos fisiopatológicos e apoiando o desenvolvimento de critérios diagnósticos com estratégias terapêuticas direcionadas.

Modelo animal de infecções associadas a implantes em camundongos

Aprenda a desenvolver modelos de infecção intracelular para Staphylococcus aureus usando cultura celular e sequenciamento de alta vazão. Este método ajuda pesquisadores a estudar mecanismos bacterianos e otimizar estratégias terapêuticas.

Desenvolvimento e Avaliação de Modelos de Infecção Intracelular para Staphylococcus aureus

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