RESUMO
Skin/soft tissue infections (SSTIs) caused by methicillin-resistant Staphylococcus aureus (MRSA) pose a major healthcare burden. Distinct inflammatory and resolution phases comprise the host immune response to SSTIs. Resolution is a myeloid PPARγ-dependent anti-inflammatory phase that is essential for the clearance of MRSA. However, the signals activating PPARγ to induce resolution remain unknown. Here, we demonstrate that myeloid glucose transporter 1 (GLUT-1) is essential for the onset of resolution. MRSA-challenged macrophages are unsuccessful in generating an oxidative burst or immune radicals in the absence of GLUT-1 due to a reduction in the cellular NADPH pool. This translates in vivo as a significant reduction in lipid peroxidation products required for the activation of PPARγ in MRSA-infected mice lacking myeloid GLUT-1. Chemical induction of PPARγ during infection circumvents this GLUT-1 requirement and improves resolution. Thus, GLUT-1-dependent oxidative burst is essential for the activation of PPARγ and subsequent resolution of SSTIs.
Assuntos
Transportador de Glucose Tipo 1 , Staphylococcus aureus Resistente à Meticilina , Infecções dos Tecidos Moles , Animais , Transportador de Glucose Tipo 1/metabolismo , Transportador de Glucose Tipo 1/genética , Camundongos , Infecções dos Tecidos Moles/microbiologia , Infecções dos Tecidos Moles/metabolismo , Infecções dos Tecidos Moles/patologia , PPAR gama/metabolismo , Infecções Cutâneas Estafilocócicas/microbiologia , Infecções Cutâneas Estafilocócicas/metabolismo , Infecções Cutâneas Estafilocócicas/patologia , Infecções Cutâneas Estafilocócicas/tratamento farmacológico , Camundongos Endogâmicos C57BL , Macrófagos/metabolismo , Macrófagos/microbiologia , Infecções Estafilocócicas/metabolismo , Infecções Estafilocócicas/microbiologiaRESUMO
The skin needs to balance tolerance of colonizing microflora with rapid detection of potential pathogens. Flexible response mechanisms would seem most suitable to accommodate the dynamic challenges of effective antimicrobial defense and restoration of tissue homeostasis. Here, we dissected macrophage-intrinsic mechanisms and microenvironmental cues that tune macrophage signaling in localized skin infection with the colonizing and opportunistic pathogen Staphylococcus aureus. Early in skin infection, the cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF) produced by γδ T cells and hypoxic conditions within the dermal microenvironment diverted macrophages away from a homeostatic M-CSF- and hypoxia-inducible factor 1α (HIF-1α)-dependent program. This allowed macrophages to be metabolically rewired for maximal inflammatory activity, which requires expression of Irg1 and generation of itaconate, but not HIF-1α. This multifactorial macrophage rewiring program was required for both the timely clearance of bacteria and for the provision of local immune memory. These findings indicate that immunometabolic conditioning allows dermal macrophages to cycle between antimicrobial activity and protection against secondary infections.
Assuntos
Macrófagos , Infecções Cutâneas Estafilocócicas , Humanos , Citocinas/metabolismo , Transdução de Sinais , Infecções Cutâneas Estafilocócicas/metabolismoRESUMO
Langerhans cells (LCs) reside in the epidermis where they are poised to mount an antimicrobial response against microbial pathogens invading from the outside environment. To elucidate potential pathways by which LCs contribute to host defense, we mined published LC transcriptomes deposited in GEO and the scientific literature for genes that participate in antimicrobial responses. Overall, we identified 31 genes in LCs that encode proteins that contribute to antimicrobial activity, ten of which were cross-validated in at least two separate experiments. Seven of these ten antimicrobial genes encode chemokines, CCL1, CCL17, CCL19, CCL2, CCL22, CXCL14 and CXCL2, which mediate both antimicrobial and inflammatory responses. Of these, CCL22 was detected in seven of nine transcriptomes and by PCR in cultured LCs. Overall, the antimicrobial genes identified in LCs encode proteins with broad antibacterial activity, including against Staphylococcus aureus, which is the leading cause of skin infections. Thus, this study illustrates that LCs, consistent with their anatomical location, are programmed to mount an antimicrobial response against invading pathogens in skin.
Assuntos
Peptídeos Antimicrobianos/genética , Epiderme/metabolismo , Células de Langerhans/metabolismo , Infecções Cutâneas Estafilocócicas/genética , Staphylococcus aureus/patogenicidade , Transcriptoma , Células Cultivadas , Bases de Dados Genéticas , Epiderme/imunologia , Epiderme/microbiologia , Perfilação da Expressão Gênica , Interações Hospedeiro-Patógeno , Humanos , Células de Langerhans/imunologia , Células de Langerhans/microbiologia , Infecções Cutâneas Estafilocócicas/imunologia , Infecções Cutâneas Estafilocócicas/metabolismo , Infecções Cutâneas Estafilocócicas/microbiologia , Staphylococcus aureus/imunologiaRESUMO
The preparation of ointments from natural compounds is essential for accelerating infected wounds. This study investigated the effects of topical uses of gold nanoparticles (Au)/perlite (Au/Perl) nanocomposites (NCs) by the help of Urtica dioica extract and its chitosan-capped derivative (Chit) on methicillin-resistant Staphylococcus aureus (MRSA)-infected wound healing in a mouse model. Furthermore, Au/Perl/Chit nanocomposite was prepared using protonated chitosan solution. The physicochemical properties of the as-synthesized nanocomposites were also investigated. The effects of Au/Perl/Chit NC were assessed by antibacterial, histopathological parameters as well as molecular evaluations. Then, they were compared with synthetic agent of mupirocin. The results revealed that Au/Perl NC was mesoporous and spherical in a range of 13-15 nm. Topical administration of Au/Perl/Chit ointment accelerated wound healing by reducing bacteria colonization and wound rate enhancing collagen biosynthesis and re-epithelialization, the expressions of IL-10, PI3K, AKT, bFGF, and COL1A genes, which is in agreement with the obtained results for mupirocin. In conclusion, the results strongly demonstrated that administration of ointments prepared from Au/Perl and Au/Perl/Chit nanocomposites stimulates MRSA-infected wound healing by decreasing the length of healing time and regulating PI3K/AKT/bFGF signaling pathway and is a promising candidate in stimulating MRSA-infected wound regeneration.
Assuntos
Óxido de Alumínio/farmacologia , Antibacterianos/farmacologia , Antioxidantes/farmacologia , Quitosana/farmacologia , Compostos de Ouro/farmacologia , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Dióxido de Silício/farmacologia , Pele/efeitos dos fármacos , Infecções Cutâneas Estafilocócicas/tratamento farmacológico , Urtica dioica/metabolismo , Cicatrização/efeitos dos fármacos , Óxido de Alumínio/metabolismo , Animais , Antibacterianos/metabolismo , Antioxidantes/metabolismo , Proliferação de Células/efeitos dos fármacos , Quitosana/análogos & derivados , Quitosana/metabolismo , Modelos Animais de Doenças , Composição de Medicamentos , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Fibroblastos/microbiologia , Fibroblastos/patologia , Compostos de Ouro/metabolismo , Química Verde , Masculino , Staphylococcus aureus Resistente à Meticilina/patogenicidade , Camundongos , Camundongos Endogâmicos BALB C , Células NIH 3T3 , Nanopartículas , Nanotecnologia , Transdução de Sinais , Dióxido de Silício/metabolismo , Pele/metabolismo , Pele/microbiologia , Pele/patologia , Infecções Cutâneas Estafilocócicas/metabolismo , Infecções Cutâneas Estafilocócicas/microbiologia , Infecções Cutâneas Estafilocócicas/patologia , Fatores de TempoRESUMO
Pore-forming toxins (PFTs), the most common virulence proteins, are promising therapeutic keys in bacterial infections. CAL02, consisting of sphingomyelin (Sm) and containing a maximum ratio of cholesterol (Ch), has been applied to sequester PFTs. However, Sm, a saturated phospholipid, leads to structural rigidity of the liposome, which does not benefit PFT combination. Therefore, in order to decrease the membrane rigidity and improve the fluidity of liposomes, we have introduced an unsaturated phospholipid, phosphatidylcholine (Pc), to the saturated Sm. In this report, a soft nanoliposome (called CSPL), composed of Ch, Sm and Pc, was artificially prepared. In order to further improve its antibacterial effect, vancomycin (Van) was loaded into the hydrophilic core of CSPL, where Van can be released radically at the infectious site through transmembrane pores formed by the PFTs in CSPL. This soft Van@CSPL nanoliposome with detoxification/drug release was able to inhibit the possibility of antibiotic resistance and could play a better role in treating severe invasive infections in mice.
Assuntos
Antibacterianos/metabolismo , Colesterol/metabolismo , Substâncias Explosivas/metabolismo , Nanopartículas/metabolismo , Esfingomielinas/metabolismo , Infecções Cutâneas Estafilocócicas/metabolismo , Animais , Antibacterianos/administração & dosagem , Anti-Infecciosos/administração & dosagem , Anti-Infecciosos/metabolismo , Colesterol/administração & dosagem , Substâncias Explosivas/administração & dosagem , Feminino , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Lipossomos , Camundongos , Nanopartículas/administração & dosagem , Esfingomielinas/administração & dosagem , Infecções Cutâneas Estafilocócicas/tratamento farmacológico , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/metabolismo , Virulência/efeitos dos fármacos , Virulência/fisiologiaRESUMO
Perforin-2 (P-2) is an antimicrobial protein with unique properties to kill intracellular bacteria. Gamma delta (GD) T cells, as the major T cell population in epithelial tissues, play a central role in protective and pathogenic immune responses in the skin. However, the tissue-specific mechanisms that control the innate immune response and the effector functions of GD T cells, especially the cross-talk with commensal organisms, are not very well understood. We hypothesized that the most prevalent skin commensal microorganism, Staphylococcus epidermidis, may play a role in regulating GD T cell-mediated cutaneous responses. We analyzed antimicrobial protein P-2 expression in human skin at a single cell resolution using an amplified fluorescence in situ hybridization approach to detect P-2 mRNA in combination with immunophenotyping. We show that S. epidermidis activates GD T cells and upregulates P-2 in human skin ex vivo in a cell-specific manner. Furthermore, P-2 upregulation following S. epidermidis stimulation correlates with increased ability of skin cells to kill intracellular Staphylococcus aureus. Our findings are the first to reveal that skin commensal bacteria induce P-2 expression, which may be utilized beneficially to modulate host innate immune responses and protect from skin infections.
Assuntos
Imunidade Inata , Proteínas Citotóxicas Formadoras de Poros/metabolismo , Receptores de Antígenos de Linfócitos T gama-delta/metabolismo , Infecções Cutâneas Estafilocócicas/imunologia , Infecções Cutâneas Estafilocócicas/metabolismo , Staphylococcus epidermidis/imunologia , Linfócitos T/imunologia , Linfócitos T/metabolismo , Biomarcadores , Citocinas/metabolismo , Citotoxicidade Imunológica , Fibroblastos/metabolismo , Regulação da Expressão Gênica , Interações Hospedeiro-Patógeno/genética , Interações Hospedeiro-Patógeno/imunologia , Humanos , Imunomodulação , Mediadores da Inflamação/metabolismo , Queratinócitos/imunologia , Queratinócitos/metabolismo , Ativação Linfocitária/genética , Ativação Linfocitária/imunologia , Contagem de Linfócitos , Proteínas Citotóxicas Formadoras de Poros/genética , Infecções Cutâneas Estafilocócicas/microbiologiaRESUMO
OBJECTIVE: The skin acts as a mechanical and protective barrier against viral, fungal, and bacterial infections. Skin conditions such as atopic dermatitis and psoriasis are characterized by alterations of the skin barrier, often caused by injury and by bacterial infections. In the last years, non-pharmacological interventions have gained great importance in epidermis-related diseases. Xyloglucan (XG) is a polysaccharide that possesses a "mucin-like" molecular structure that confers mucoadhesive properties, allowing XG-containing formulations to act as a protective barrier for the management of different diseases. Moreover, there is also increasing interest in the use of proteins due to their film-forming features. This study aimed to evaluate the barrier-protective properties of a product containing XG and pea protein (PP) in an in vitro model, assessing its effects on the membrane permeability of keratinocytes infected by Staphylococcus aureus. METHODS: HaCaT keratinocytes were pretreated with XG and PP for 3 h and then infected with S. aureus cells (106 bacteria/well) at a multiplicity of infection of 10 for 1 h. The number of bacterial colonies and membrane integrity were measured, respectively. RESULTS: We observed that pretreatment with XG and PP in human HaCaT keratinocytes infected with S. aureus significantly increased trans-epithelial electrical resistance (a marker of skin barrier function) measurement, reduced lucifer yellow (a marker of membrane integrity) permeation across the monolayer, and released lactate dehydrogenase (a marker of tissue damage). Moreover, XG and PP pretreatment was able to reduce bacterial adherence, avoiding S. aureus infection. CONCLUSION: In summary, we demonstrated that the product containing XG and PP was able to maintain barrier permeability preserving its integrity, and therefore, it can be considered as an interesting approach for the management of epidermis-related diseases.
Assuntos
Glucanos/farmacologia , Queratinócitos/efeitos dos fármacos , Proteínas de Ervilha/farmacologia , Pele/efeitos dos fármacos , Infecções Cutâneas Estafilocócicas/tratamento farmacológico , Staphylococcus aureus/efeitos dos fármacos , Xilanos/farmacologia , Aderência Bacteriana/efeitos dos fármacos , Células HaCaT , Humanos , Queratinócitos/metabolismo , Queratinócitos/microbiologia , Permeabilidade , Pele/metabolismo , Pele/microbiologia , Infecções Cutâneas Estafilocócicas/metabolismo , Infecções Cutâneas Estafilocócicas/microbiologia , Staphylococcus aureus/patogenicidadeRESUMO
With the effectiveness of antimicrobials declining as antimicrobial resistance continues to threaten public health, we must look to alternative strategies for the treatment of infections. In this study, we investigated an innovative, drug-free, dual-wavelength irradiation approach that combines 2 wavelengths of light, 460 nm and 405 nm, against methicillin-resistant Staphylococcus aureus (MRSA). MRSA was initially irradiated with 460-nm light (90-360 J/cm2) and subsequently irradiated with aliquots of 405-nm light (54-324 J/cm2). For in vivo studies, mouse skin was abraded and infected with approximately 107 CFUs of MRSA and incubated for 3 hours before irradiating with 460 nm (360 J/cm2) and 405 nm (342 J/cm2). Naive mouse skin was also irradiated to investigate apoptosis. We found that staphyloxanthin, the carotenoid pigment in MRSA cells, promoted resistance to the antimicrobial effects of 405-nm light. In addition, we found that the photolytic effect of 460-nm light on staphyloxanthin attenuated resistance of MRSA to 405-nm light killing. Irradiation of 460 nm alone did not elicit any antimicrobial effect on MRSA. In a proof-of-principle mouse skin abrasion infection model, we observed significant killing of MRSA using the dual-wavelength irradiation approach. However, when either wavelength of light was administered alone, no significant decrease in bacterial viability was observed. Moreover, exposure of the dual-wavelength irradiation to naive mouse skin did not result in any visible apoptosis. In conclusion, a dual-wavelength irradiation strategy may offer an innovative, effective, and safe approach for the treatment of skin infections caused by MRSA.
Assuntos
Staphylococcus aureus Resistente à Meticilina/crescimento & desenvolvimento , Fototerapia , Infecções Cutâneas Estafilocócicas , Animais , Modelos Animais de Doenças , Infecções Cutâneas Estafilocócicas/metabolismo , Infecções Cutâneas Estafilocócicas/patologia , Infecções Cutâneas Estafilocócicas/terapiaRESUMO
Staphylococcus aureus small colony variants (SCVs) are frequently associated with chronic infection, yet they lack expression of many virulence determinants associated with the pathogenicity of wild-type strains. We found that both wild-type S. aureus and a ΔhemB SCV prototype potently activate glycolysis in host cells. Glycolysis and the generation of mitochondrial reactive oxygen species were sufficient to induce necroptosis, a caspase-independent mechanism of host cell death that failed to eradicate S. aureus and instead promoted ΔhemB SCV pathogenicity. To support ongoing glycolytic activity, the ΔhemB SCV induced over a 100-fold increase in the expression of fumC, which encodes an enzyme that catalyses the degradatin of fumarate, an inhibitor of glycolysis. Consistent with fumC-dependent depletion of local fumarate, the ΔhemB SCV failed to elicit trained immunity and protection from a secondary infectious challenge in the skin. The reliance of the S. aureus SCV population on glycolysis accounts for much of its role in the pathogenesis of S. aureus skin infection.
Assuntos
Imunomodulação , Infecções Cutâneas Estafilocócicas/metabolismo , Infecções Cutâneas Estafilocócicas/patologia , Staphylococcus aureus/patogenicidade , Animais , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Células Cultivadas , Fumaratos/metabolismo , Regulação Bacteriana da Expressão Gênica , Glicólise , Humanos , Evasão da Resposta Imune , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Mutação , Necroptose/genética , Espécies Reativas de Oxigênio/metabolismo , Infecções Cutâneas Estafilocócicas/imunologia , Infecções Cutâneas Estafilocócicas/microbiologia , Staphylococcus aureus/genética , Staphylococcus aureus/crescimento & desenvolvimento , Staphylococcus aureus/metabolismo , Células THP-1RESUMO
In vivo whole-animal optical (bioluminescence and fluorescence) imaging of Staphylococcus aureus infections has provided the opportunity to noninvasively and longitudinally monitor the dynamics of the bacterial burden and ensuing host immune responses in live anesthetized animals. Herein, we describe several different mouse models of S. aureus skin infection, skin inflammation, incisional/excisional wound infections, as well as mouse and rabbit models of orthopedic implant infection, which utilized this imaging technology. These animal models and imaging methodologies provide insights into the pathogenesis of these infections and innate and adaptive immune responses, as well as the preclinical evaluation of diagnostic and treatment modalities. Noninvasive approaches to investigate host-pathogen interactions are extremely important as virulent community-acquired methicillin-resistant S. aureus strains (CA-MRSA) are spreading through the normal human population, becoming more antibiotic resistant and creating a serious threat to public health.
Assuntos
Staphylococcus aureus Resistente à Meticilina/metabolismo , Imagem Óptica , Infecções Cutâneas Estafilocócicas , Animais , Modelos Animais de Doenças , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Coelhos , Infecções Cutâneas Estafilocócicas/diagnóstico , Infecções Cutâneas Estafilocócicas/metabolismo , Infecções Cutâneas Estafilocócicas/patologiaRESUMO
Effective antimicrobial preparations, other than antibiotics, are important for the treatment of potentially fatal drug-resistant infections. Methicillin-resistant Staphylococcus aureus (MRSA) is one of the leading causes of hospital-acquired and post- operative infections. Fortunately, the antimicrobial properties of platelet-rich plasma (PRP) against various microorganisms enable its potential use as an alternative to conventional antibiotics. The present work was designed to evaluate the hypothesized antimicrobial activity of PRP against MRSA infected skin wounds. Six adult male dogs were divided equally into control and PRP groups. Unilateral circular full-thickness skin wounds were created then a MRSA suspension was injected locally. Treatment started at 1st week post infection with subcutaneous infiltration of autologous activated PRP every week in the PRP group and with topical application of clindamycin cream twice daily in the control group. PRP decreased wound size and significantly increased wound contractility and re-epithelization, as confirmed by histopathological and immunohistochemical findings. Also PRP treated group showed significant decrease in ROS and redox imbalance with over expression of the TNF-α and VEGFA genes that indicate angiogenesis and maximum antibacterial activity after three weeks. In conclusion, CaCl2-activated PRP exhibited antimicrobial activity against MRSA infection, which improved the infected wound healing re-epithelization and granulation tissue formation.
Assuntos
Staphylococcus aureus Resistente à Meticilina/metabolismo , Plasma Rico em Plaquetas , Infecções Cutâneas Estafilocócicas , Infecção da Ferida Cirúrgica , Cicatrização , Animais , Cães , Masculino , Infecções Cutâneas Estafilocócicas/metabolismo , Infecções Cutâneas Estafilocócicas/terapia , Infecção da Ferida Cirúrgica/metabolismo , Infecção da Ferida Cirúrgica/microbiologia , Infecção da Ferida Cirúrgica/terapiaRESUMO
Staphylococcus aureus can cause persistent infections and is known to develop persister cells in vitro. However, the in vivo significance of in vitro persisters in general is largely unclear. Here, we evaluated S. aureus stationary phase cultures and biofilm bacteria enriched in persister bacteria in comparison with actively growing log phase bacteria in terms of their ability to cause disease in a mouse skin infection model. We found that mice infected with the stationary phase and biofilm bacteria, which were enriched with persisters, produced more pronounced skin lesions that took longer to heal, and had more severe skin pathology and higher bacterial load than mice infected with log phase bacteria. Using our persistent infection mouse model, we showed that the clinically recommended treatment for recurrent S. aureus skin infection, doxycycline + rifampin, was not effective in eradicating the bacteria in mice. Analogous findings were observed in a Caenorhabditis elegans model, where stationary phase S. aureus caused greater virulence or mortality than log phase bacteria as early as two days post-infection. Our findings associate in vitro persisters and biofilm bacteria with more persistent and more severe infections and emphasize the importance of quality or metabolic status of the inoculum bacteria (persister bacteria versus growing bacteria) not just the number of bacteria in causing disease. The persistent infection mouse model we developed with persister inocula should have implications for understanding the process of disease establishment and pathogenesis, for developing persistent infection animal models, and for developing more effective treatments for chronic persistent infections in general.
Assuntos
Biofilmes/efeitos dos fármacos , Doxiciclina/farmacologia , Rifampina/farmacologia , Infecções Cutâneas Estafilocócicas/tratamento farmacológico , Staphylococcus aureus/fisiologia , Animais , Caenorhabditis elegans/microbiologia , Modelos Animais de Doenças , Feminino , Camundongos , Infecções Cutâneas Estafilocócicas/metabolismo , Infecções Cutâneas Estafilocócicas/patologiaRESUMO
Staphylococcus aureus expresses a number of cell wall-anchored proteins that mediate adhesion and invasion of host cells and tissues and promote immune evasion, consequently contributing to the virulence of this organism. The cell wall-anchored protein clumping factor B (ClfB) has previously been shown to facilitate S. aureus nasal colonization through high affinity interactions with the cornified envelope in the anterior nares. However, the role of ClfB during skin and soft tissue infection (SSTI) has never been investigated. This study reveals a novel role for ClfB during SSTIs. ClfB is crucial in determining the abscess structure and bacterial burden early in infection and this is dependent upon a specific interaction with the ligand loricrin which is expressed within the abscess tissue. Targeting ClfB using a model vaccine that induced both protective humoral and cellular responses, leads to protection during S. aureus skin infection. This study therefore identifies ClfB as an important antigen for future SSTI vaccines.
Assuntos
Adesinas Bacterianas/metabolismo , Infecções Cutâneas Estafilocócicas/microbiologia , Staphylococcus aureus/imunologia , Vacinas/imunologia , Fatores de Virulência/metabolismo , Virulência , Adesinas Bacterianas/genética , Adesinas Bacterianas/imunologia , Animais , Aderência Bacteriana , Feminino , Proteínas de Membrana/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Infecções Cutâneas Estafilocócicas/imunologia , Infecções Cutâneas Estafilocócicas/metabolismo , Vacinas/administração & dosagem , Fatores de Virulência/genética , Fatores de Virulência/imunologiaRESUMO
Staphylococcus pseudintermedius is an opportunistic and emerging zoonotic pathogen that primarily colonises the skin of dogs. Many common variants are methicillin resistant (MRSP) or multidrug resistant (MDR), and drug resistance is increasingly reported across the globe. In New Zealand, MRSP isolation remains rare in clinics. To pre-emptively inform diagnostic and antimicrobial stewardship practices, we examine isolates of S. pseudintermedius, MRSP and MDR-MRSP from New Zealand dogs using a combination of methodologies. Genetic and genomic data combined with antimicrobial susceptibility screening identify common drug-resistance profiles and their genetic determinants. We demonstrate that sensitive and specific species-level identification of S. pseudintermedius can be achieved using Bruker MALDI-TOF MS and, further, that this technique can be used to identify some common subtype variants, providing a level of categorical precision that falls somewhere between single-locus and multi-locus sequence typing. Comparative genomics analysis of global S. pseudintermedius data shows that MRSP moves frequently across the globe, but that horizontal gene transfer events resulting in the acquisition of the SCCmec cassette (responsible for beta-lactam antibiotic resistance) are infrequent. This suggests that biosecurity and surveillance in addition to antibiotic stewardship should play important roles in mitigating the risk of MRSP, especially in countries such as New Zealand where MRSP is still rare.
Assuntos
Doenças do Cão , Farmacorresistência Bacteriana Múltipla , Genômica , Resistência a Meticilina , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Infecções Cutâneas Estafilocócicas , Staphylococcus , Animais , Doenças do Cão/genética , Doenças do Cão/metabolismo , Doenças do Cão/microbiologia , Cães , Nova Zelândia , Infecções Cutâneas Estafilocócicas/genética , Infecções Cutâneas Estafilocócicas/metabolismo , Infecções Cutâneas Estafilocócicas/veterinária , Staphylococcus/genética , Staphylococcus/metabolismoRESUMO
Garcinia mangostana (mangosteen) pericarp has antibacterial effects; however, information regarding its anti-inflammatory activity in vivo is limited. The anti-inflammatory effect of G. mangostana pericarp extract against methicillin-resistant Staphylococcus aureus (MRSA)-induced superficial skin infection was investigated in mice using a tape stripping model. G. mangostana pericarp ethanolic extract (GME) and its constituent, α-mangostin, were topically administered to mice with MRSA-induced superficial skin infection. MRSA-infected wounds treated with GME were completely healed on the 10th day of the study and the number of MRSA-colonies decreased from the first day of the study, whereas α-mangostin-treated wounds never completely healed with higher numbers of MRSA colonies. The epidermis of GME-treated wounds had nearly completely regenerated, with no inflammatory cell infiltration. In contrast, α-mangostin-treated wounds exhibited neutrophil infiltration and accumulation of mast cells. MRSA-infected wounds without treatment showed high expression of TNF-α, IL-6, IL-1ß, and TLR-2 genes. In contrast, GME decreased mRNA levels, restoring expression of those genes to normal levels. Notably, α-mangostin did not down-regulate the expression of pro-inflammatory cytokines to the same extent as GME. Hence, GME is a promising alternative MRSA treatment because of its antibacterial, anti-inflammatory, and wound healing effects.
Assuntos
Anti-Inflamatórios/uso terapêutico , Garcinia mangostana , Mediadores da Inflamação/antagonistas & inibidores , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Extratos Vegetais/uso terapêutico , Infecções Cutâneas Estafilocócicas/tratamento farmacológico , Animais , Anti-Inflamatórios/isolamento & purificação , Anti-Inflamatórios/farmacologia , Mediadores da Inflamação/metabolismo , Masculino , Staphylococcus aureus Resistente à Meticilina/fisiologia , Camundongos , Camundongos Endogâmicos ICR , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/farmacologia , Infecções Estafilocócicas/tratamento farmacológico , Infecções Estafilocócicas/metabolismo , Infecções Estafilocócicas/patologia , Infecções Cutâneas Estafilocócicas/metabolismo , Infecções Cutâneas Estafilocócicas/patologia , Resultado do TratamentoRESUMO
Mast cells (MCs) are strategically distributed at barrier sites and prestore various immunocyte-recruiting cytokines, making them ideal targets for selective activation to treat peripheral infections. Here, we report that topical treatment with mastoparan, a peptide MC activator (MCA), enhances clearance of Staphylococcus aureus from infected mouse skins and accelerates healing of dermonecrotic lesions. Mastoparan functions by activating connective tissue MCs (CTMCs) via the MRGPRX2 (Mas-related G protein-coupled receptor member X2) receptor. Peripheral CTMC activation, in turn, enhances recruitment of bacteria-clearing neutrophils and wound-healing CD301b+ dendritic cells. Consistent with MCs playing a master coordinating role, MC activation also augmented migration of various antigen-presenting dendritic cells to draining lymph nodes, leading to stronger protection against a second infection challenge. MCAs therefore orchestrate both the innate and adaptive immune arms, which could potentially be applied to combat peripheral infections by a broad range of pathogens.
Assuntos
Mastócitos/imunologia , Mastócitos/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Receptores Acoplados a Proteínas G/metabolismo , Receptores de Neuropeptídeos/metabolismo , Infecções Cutâneas Estafilocócicas/imunologia , Infecções Cutâneas Estafilocócicas/metabolismo , Imunidade Adaptativa/efeitos dos fármacos , Administração Tópica , Animais , Células Dendríticas/imunologia , Células Dendríticas/metabolismo , Feminino , Células HEK293 , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/administração & dosagem , Peptídeos e Proteínas de Sinalização Intercelular/uso terapêutico , Masculino , Mastócitos/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Proteínas do Tecido Nervoso/genética , Neutrófilos/imunologia , Neutrófilos/metabolismo , Receptores Acoplados a Proteínas G/genética , Receptores de Neuropeptídeos/genética , Infecções Cutâneas Estafilocócicas/tratamento farmacológico , Infecções Cutâneas Estafilocócicas/microbiologia , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/crescimento & desenvolvimento , Transfecção , Venenos de Vespas/administração & dosagem , Venenos de Vespas/uso terapêutico , Cicatrização/efeitos dos fármacos , Cicatrização/imunologiaRESUMO
Staphylococcus epidermidis is an abundant skin commensal capable of activating cutaneous defense responses, such as induction of cytokines and antimicrobial peptides. To permanently colonize human skin and prevent inflammation S. epidermidis needs to control the induction of host defense mediators. We report here that S. epidermidis induces expression of the host regulator protein A20 in human keratinocytes, thereby controlling expression and release of interleukin-1 beta. siRNA-mediated knockdown of A20 expression strongly enhanced the induction of interleukin-1 beta gene expression and protein release in keratinocytes stimulated with S. epidermidis. Furthermore, siRNA-mediated knockdown of A20 resulted in enhanced gene expression and secretion of the antimicrobial peptide human beta-defensin-2 in keratinocytes facing S. epidermidis. Mechanistically, A20 negatively controlled S. epidermidis-induced activation of the transcription factor NF-kappaB. Together, these data indicate that S. epidermidis exploits A20 to attenuate cutaneous defense responses, which may help S. epidermidis to persist on human skin.
Assuntos
Interleucina-1beta/metabolismo , Queratinócitos/microbiologia , Pele/microbiologia , Infecções Cutâneas Estafilocócicas/microbiologia , Staphylococcus epidermidis/patogenicidade , Proteína 3 Induzida por Fator de Necrose Tumoral alfa/metabolismo , beta-Defensinas/metabolismo , Células Cultivadas , Interações Hospedeiro-Patógeno , Humanos , Interleucina-1beta/genética , Queratinócitos/metabolismo , NF-kappa B/metabolismo , Transdução de Sinais , Pele/metabolismo , Infecções Cutâneas Estafilocócicas/genética , Infecções Cutâneas Estafilocócicas/metabolismo , Proteína 3 Induzida por Fator de Necrose Tumoral alfa/genética , Regulação para Cima , beta-Defensinas/genéticaRESUMO
Abuse of antibiotics and their residues in the environment results in the emergence and prevalence of drug-resistant bacteria and leads to serious health problems. Herein, a photon-controlled antibacterial platform that can efficiently kill drug-resistant bacteria and avoid the generation of new bacterial resistance was designed by encapsulating black phosphorus quantum dots (BPQDs) and pharmaceuticals inside a thermal-sensitive liposome. The antibacterial platform can release pharmaceuticals in a spatial-, temporal-, and dosage-controlled fashion because the BPQDs can delicately generate heat under near-infrared light stimulation to disrupt the liposome. This user-defined delivery of drug can greatly reduce the antibiotic dosage, thus avoiding the indiscriminate use of antibiotics and preventing the generation of superbugs. Moreover, by coupling the photothermal effect with antibiotics, this antibacterial platform achieved a synergistic photothermal-/pharmaco-therapy with significantly improved antibacterial efficiency toward drug-resistant bacteria. The antibacterial platform was further employed to treat antibiotic-resistant bacteria-caused skin abscess and it displayed excellent antibacterial activity in vivo, promising its potential clinical applications. Additionally, the antibacterial mechanism was further investigated. The developed photon-controlled antibacterial platform can open new possibilities for avoiding bacterial resistance and efficiently killing antibiotic-resistant bacteria, making it valuable in fields ranging from antiinfective therapy to precision medicine.
Assuntos
Antibacterianos , Hipertermia Induzida , Raios Infravermelhos , Staphylococcus aureus Resistente à Meticilina/crescimento & desenvolvimento , Fototerapia , Pontos Quânticos , Infecções Cutâneas Estafilocócicas , Animais , Antibacterianos/química , Antibacterianos/farmacocinética , Antibacterianos/farmacologia , Lipossomos , Camundongos , Pontos Quânticos/química , Pontos Quânticos/uso terapêutico , Infecções Cutâneas Estafilocócicas/metabolismo , Infecções Cutâneas Estafilocócicas/patologia , Infecções Cutâneas Estafilocócicas/terapiaRESUMO
Atopic dermatitis (AD) is a chronic inflammatory skin disease with a defective immunologic barrier, which is aggravated by Staphylococcus aureus (S. aureus). Epidermal growth factor (EGF) suppresses inflammation and EGF receptor inhibitors increased S. aureus colonization. Thus, we investigated the potential roles of EGF in AD, which is often aggravated by S. aureus. We determined how EGF affects the expression of inflammatory cytokines and antimicrobial peptides (AMPs) in human epidermal keratinocytes (HEKs) treated with heat-inactivated S. aureus (HKSA) in vitro and 2,4-dinitrochlorobenzene-induced AD-like skin lesions in Nc/Nga mice. HKSA increased IL-6 and NFκB expression; EGF treatment had the opposite effect. EGF increased human ß defensin-2 expression in HEKs and murine ß defensin-3 in mice. In mice, both EGF and pimecrolimus groups showed less erythema with significantly reduced inflammation and decreased expression of thymic stromal lymphopoietin. EGF relieved S. aureus-induced inflammation and AD-like skin lesions in Nc/Nga mice. Therefore, EGF could be a potential topical treatment for AD.
Assuntos
Dermatite Atópica , Transdução de Sinais/efeitos dos fármacos , Infecções Cutâneas Estafilocócicas , Staphylococcus aureus/metabolismo , Animais , Dermatite Atópica/tratamento farmacológico , Dermatite Atópica/metabolismo , Dermatite Atópica/microbiologia , Dermatite Atópica/patologia , Fator de Crescimento Epidérmico/metabolismo , Fator de Crescimento Epidérmico/farmacologia , Humanos , Inflamação/tratamento farmacológico , Inflamação/metabolismo , Inflamação/patologia , Masculino , Camundongos , Infecções Cutâneas Estafilocócicas/tratamento farmacológico , Infecções Cutâneas Estafilocócicas/metabolismo , Infecções Cutâneas Estafilocócicas/patologiaRESUMO
Inflammation at a wound site is essential for preventing infection. However, misregulated inflammation leads to pathologies of the healing process, including chronic non-healing wounds and scarring. MicroRNAs (miRNAs) are key regulators of the inflammatory response and tissue repair, acting by translational processing of target mRNAs. In the final step of miRNA processing, Argonaute 2 (Ago2)-bound mature miRNA complexes bind to target mRNAs and inhibit their translation. A variety of wound healing-related miRNAs have been identified and their misregulation likely contributes to wound pathologies, including scarring and chronic healing. Recently, we have developed an Ago2-bound mature miRNA purification system that uses Ago2 antibody to analyze the expression of miRNAs from wound tissues by microarray and next generation sequencing. We have identified several wound inflammation-related miRNAs via Ago2-target immunoprecipitation assays and next generation sequencing of wound tissues from wild-type and PU.1 knockout mice, which exhibit no inflammatory response because of a lack of immune cell lineages. We demonstrated that miR-142, an identified inflammation-related miRNA, is essential role for neutrophilic chemotaxis via inhibition of small GTPase translation; its misregulation leads to susceptibility to infection against Staphylococcus aureus at skin wound sites. In this review, we summarize recent advances of miRNA studies in skin wound healing, introduce our miRNA purification system using an immunoprecipitation assay method, and discuss the function of miR-142 in skin wound healing.