RESUMO
Chromoblastomycosis (CBM) is a neglected human disease, caused by different species of pigmented dematiaceous fungi that cause subcutaneous infections. This disease has been considered an occupational disease, occurring among people working in the field of agriculture, particularly in low-income countries. In 1914, the first case of CBM was described in Brazil, and although efforts have been made, few scientific and technological advances have been made in this area. In the field of fungi and host cell relationship, a very reduced number of antigens were characterized, but available data suggest that ectoantigens bind to the cell membrane of host cells and modulate the phagocytic, immunological, and microbicidal responses of immune cells. Furthermore, antigens cleave extracellular proteins in tissues, allowing fungi to spread. On the contrary, if phagocytic cells are able to present antigens in MHC molecules to T lymphocytes in the presence of costimulation and IL-12, a Th1 immune response will develop and a relative control of the disease will be observed. Despite knowledge of the resistance and susceptibility in CBM, up to now, no effective vaccines have been developed. In the field of chemotherapy, most patients are treated with conventional antifungal drugs, such as itraconazole and terbinafine, but these drugs exhibit limitations, considering that not all patients heal cutaneous lesions. Few advances in treatment have been made so far, but one of the most promising ones is based on the use of immunomodulators, such as imiquimod. Data about a standard treatment are missing in the medical literature; part of it is caused by the existence of a diversity of etiologic agents and clinical forms. The present review summarizes the advances made in the field of CBM related to the diversity of pathogenic species, fungi and host cell relationship, antigens, innate and acquired immunity, clinical forms of CBM, chemotherapy, and diagnosis.
Assuntos
Cromoblastomicose/imunologia , Interações entre Hospedeiro e Microrganismos/imunologia , Imunidade/imunologia , Animais , Antifúngicos/imunologia , Antígenos/imunologia , Humanos , Fatores Imunológicos/imunologiaRESUMO
Usually caused by Candida albicans, buccal candidiasis begins with the morphological transition between yeast and hyphal cells. Over time and without the correct treatment, it can be disseminated through the bloodstream becoming a systemic infection with high mortality rates. C. albicans already shows resistance against antifungals commonly used in treatments. Therefore, the search for new drugs capable of overcoming antifungal resistance is essential. Histatin 5 (Hst5) is an antimicrobial peptide of the Histatin family, that can be found naturally in human saliva. This peptide presents high antifungal activity against C. albicans. However, Hst5 action can be decreased for interaction with enzymes and metal ions present in the oral cavity. The current work aims to bring a brief review of relevant aspects of the pathogenesis and resistance mechanisms already reported for C. albicans. In addition, are also reported here the main immune responses of the human body and the most common antifungal drugs. Finally, the most important aspects regarding Histatin 5 and the benefits of its interaction with metals are highlighted. The intention of this review is to show the promising use of Hst5 metallopeptides in the development of effective drugs.
Assuntos
Antifúngicos/imunologia , Candida albicans , Candidíase , Farmacorresistência Fúngica , Histatinas/imunologia , Saliva/imunologia , Animais , Candida albicans/imunologia , Candida albicans/fisiologia , Candidíase/tratamento farmacológico , Candidíase/imunologia , HumanosRESUMO
Resumo: A epidemiologia das infecções causadas por Candida sofreu mudanças nos últimos anos. Embora Candida albicans ainda seja o principal patógeno causador de candidíase, relatos recentes reportam o aumento da incidência de infecções causadas por espécies de Candida não-albicans. A toxicidade dos fármacos antifúngicos utilizados juntamente com o desenvolvimento da resistência tem se tornado um sério problema clínico. Com isso, atualmente existe a urgência na descoberta de novos compostos com atividades antifúngicas. Objetivos: avaliar a atividade antifúngica do fármaco Cloridrato de Verapamil, determinar a Concentração Inibitória Mínima (CIM) e a Concentração Fungicida Mínima (CFM), frente à Candida albicans ATCC 18804, Candida krusei ATCC 6258, Candida parapsilosis ATCC 90018 e Candida glabrata ATCC 9030. Além de avaliar a citotoxicidade do fármaco em queratinócitos humanos (HaCaT). Metodologia: a determinação da CIM foi realizada pela técnica de microdiluição de acordo com The European Committee on Antimicrobial Susceptibility Testing (EUCAST). A CFM foi determinada por plaqueamento em ágar Sabouraud de alíquotas provenientes das diluições do ensaio de microdiluição. A avaliação da citotoxicidade foi realizada pela técnica de redução da resazurina, sendo possível avaliar a atividade mitocondrial das células HaCaT. Resultados: o fármaco Cloridrato de Verapamil demonstrou atividade antifúngica contra as quatro espécies patogênicas de Candida, com o valor de CIM de 1250 µM; valor de CFM maior que 1250 µM, sendo assim este fármaco foi considerado fungistático. Além disso, o Cloridrato de Verapamil não apresentou citotoxicidade nas concentrações avaliadas no estudo, pois, a redução de células viáveis nas concentrações mais elevadas não ultrapassa 30%. Com relação à redução de biomassa em biofilme de Candida, após tratamento com Cloridrato de Verapamil, houve redução (de 10 a 20%) para as quatro espécies de Candida em estudo, quando utilizadas concentrações de fármaco correspondentes a CIM; quando utilizadas concentrações de fármaco correspondentes a cinco vezes o valor de CIM, houve um aumento significativo na redução de biomassa (de 25% a 60%) em biofilme formado pelas quatro espécies de Candida. Conclusão: o fármaco Cloridrato de Verapamil apresentou atividade antifúngica para Candida albicans e Candida não-albicans, sendo considerado um fármaco fungistático, além de não apresentar citotoxicidade em queratinócitos humanos, e demonstrar atividade na redução de biomassa em biofilme formado pelas quatro espécies de Candida. Demais estudos são necessários para verificar a ação desse fármaco em diferentes mecanismos de virulência frente a células de Candida spp. (AU)
Abstract: The epidemiology of Candida infections has changed in recent years. Although Candida albicans is still the main pathogen causing candidiasis, recent reports have reported an increase in the incidence of infections caused by nonalbicans Candida species. The toxicity of the antifungal drugs used together with the development of resistance has become a serious clinical problem. With this, there is now an urgency in the discovery of new compounds with antifungal activities. Objectives: To evaluate the antifungal activity of the drug Verapamil Hydrochloride, to determine the Minimum Inhibitory Concentration (MIC) and the Minimum Fungicidal Concentration (CFM) against Candida albicans ATCC 18804, Candida krusei ATCC 6258, Candida parapsilosis ATCC 90018 and Candida glabrata ATCC 9030 In addition to evaluating the cytotoxicity of the drug in human keratinocytes (HaCaT). Methodology: MIC determination was performed by the microdilution technique according to The European Committee on Antimicrobial Susceptibility Testing (EUCAST). The CFM was determined by plating on Sabouraud agar from aliquots from the dilutions of the microdilution assay. The evaluation of cytotoxicity was performed using the resazurin reduction technique, and it was possible to evaluate the mitochondrial activity of HaCaT cells. Results: The drug Verapamil Hydrochloride demonstrated antifungal activity against the four pathogenic species of Candida, with MIC value of 1250 µM; CFM value greater than 1250 µM, so this drug was considered fungistatic. In addition, Verapamil Hydrochloride did not present cytotoxicity in the units evaluated in the study, because the reduction of viable cells in the most cells is not exceeded by 30%. Regarding the biomass reduction in Candida biofilm, after treatment with Verapamil Hydrochloride, there was a reduction (from 10 to 20%) for the four Candida species in study, when the use of drug corresponding to MIC; when the use of drug corresponding to 5 times of MIC, there was a significant increase in the biomass reduction (from 25% to 60%) in the biofilm molded by the four Candida species. Conclusion: Thus, the drug Verapamil Hydrochloride led to the antifungal activity of Candida albicans and non-albicans Candida species, being considered a fungicidal drug, besides not presenting cytotoxicity in human serotonizers, and were submitted to biomass reduction in biofilm molded by four Candida species. More studies are needed to verify the action of the drug on different mechanisms of virulence against Candida spp cells(AU)
Assuntos
Humanos , Candida , Antifúngicos/imunologiaRESUMO
Cryptococcus neoformans is an encapsulated fungal pathogen that causes cryptococcosis, which is a major opportunistic infection in immunosuppressed individuals. Mammalian ß-galactoside-binding protein Galectin-3 (Gal-3) modulates the host innate and adaptive immunity, and plays significant roles during microbial infections including some fungal diseases. Here we show that this protein plays a role also in C. neoformans infection. We find augmented Gal-3 serum levels in human and experimental infections, as well as in spleen, lung, and brain tissues of infected mice. Gal-3-deficient mice are more susceptible to cryptococcosis than WT animals, as demonstrated by the higher fungal burden and lower animal survival. In vitro experiments show that Gal-3 inhibits fungal growth and exerts a direct lytic effect on C. neoformans extracellular vesicles (EVs). Our results indicate a direct role for Gal-3 in antifungal immunity whereby this molecule affects the outcome of C. neoformans infection by inhibiting fungal growth and reducing EV stability, which in turn could benefit the host.
Assuntos
Antifúngicos/imunologia , Antifúngicos/farmacologia , Criptococose/tratamento farmacológico , Criptococose/imunologia , Cryptococcus neoformans/efeitos dos fármacos , Galectina 3/imunologia , Galectina 3/farmacologia , Imunidade Adaptativa , Animais , Cápsulas Bacterianas/efeitos dos fármacos , Proteínas Sanguíneas , Encéfalo/imunologia , Criptococose/microbiologia , Cryptococcus neoformans/crescimento & desenvolvimento , Cryptococcus neoformans/imunologia , Citocinas/metabolismo , Modelos Animais de Doenças , Galectina 3/sangue , Galectina 3/genética , Galectinas , Expressão Gênica , Humanos , Pulmão/imunologia , Macrófagos/efeitos dos fármacos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Baço/imunologiaRESUMO
The chapter reviews methods utilized for the isolation and characterization of a promising immunogen candidate, aiming at a human vaccine against paracoccidioidomycosis. Peptide P10 carries a T-CD4+ epitope and was identified as an internal sequence of the major diagnostic antigen known as gp43 glycoprotein. It successfully treated massive intratracheal infections by virulent Paracoccidioides brasiliensis in combination with chemotherapy.An introduction about the systemic mycosis was found essential to understand the various options that were considered to design prophylactic and therapeutic vaccine protocols using peptide P10.
Assuntos
Blastomyces/imunologia , Vacinas Fúngicas/imunologia , Paracoccidioidomicose/imunologia , Paracoccidioidomicose/prevenção & controle , Vacinas de Subunidades Antigênicas/imunologia , Animais , Antifúngicos/imunologia , Antígenos de Fungos/química , Antígenos de Fungos/imunologia , Antígenos de Fungos/isolamento & purificação , Citocinas/metabolismo , Modelos Animais de Doenças , Proteínas Fúngicas , Humanos , Imunização , Pulmão/imunologia , Pulmão/metabolismo , Pulmão/patologia , Camundongos , Paracoccidioidomicose/tratamento farmacológico , Peptídeos/química , Peptídeos/imunologia , Peptídeos/isolamento & purificação , Proteômica/métodosRESUMO
O objetivo deste trabalho foi sintetizar e caracterizar nanopartículas de sílica recobertas por prata, e avaliar a influência quando incorporadas à resina acrílica quimicamente ativada, ao reembasador macio, e a um glaze, em suas características microbiológicas, microestruturais e mecânicas. Foi confeccionada, pelo método de hidrólise e condensação controlada (método de Stober), uma solução contendo nanopartículas de sílica revestidas por nanopartículas de prata em duas proporções, 10 milimols e 30 milimols, onde as mesmas foram caracterizadas. As nanopartículas de sílica incorporadas com a prata foram analisadas por energia dispersiva de raios-X integrado (EDS), microscopia eletrônica de varredura (MEV), e espalhamento de luz dinâmico (DLS). Para análise microbiológica foram confeccionadas 10 amostras cilíndricas (2mmx10mm), em resina acrílica quimicamente ativada onde as partículas foram incorporadas em duas concentrações: 2.5% e 5% com as duas molaridades diferentes 10 mM e 30 mM. Em outra situação foram confeccionadas 10 amostras cilíndricas (2mmx10mm) de reembasador macio, com concentrações de 2,5% e 5% com as duas molaridades diferentes 10 mM e 30 mM, no terceiro caso as partículas foram acrescentadas a um glaze nas duas concentrações e molaridades e aplicada sob uma amostra de reembasador macio SoftConfort (Dencril) a base de resina acrílica. Uma suspensão de Candida albicans e Estreptococos mutans foi utilizada para análise de concentração inibitória mínima. Amostras retangulares em resina acrílica (n=6) de 30x10x3mm, foram confeccionadas para realização do teste de resistência a flexão de três pontos de ensaios mecânicos EMIC (Modelo DL-1000, EMIC Equipamentos e Sistemas LTDA., São José dos Pinhais - PR Brasil), com velocidade de 1mm/min. Amostras sem a presença de nanopartículas foram confeccionadas como grupo controle. A sílica inicialmente apresentava-se como um pó de coloração branca e fina, após a sua caracterização com as partículas de Nitrato de Prata, houve alteração da coloração para um tom amarelado, aumento da sua densidade e do tamanho de suas nanopartículas. Os resultados de ação antimicrobiana foram positivos para as amostras de reembasador macio e glaze com 5% e 30 mMol de nanopartículas, já no ensaio mecânico não houve diferença significante estatisticamente entre os grupos(AU)
The aim of this work was to synthesize and characterize silver - coated silica nanoparticles, and to evaluate the influence when incorporated into chemically activated acrylic resin, soft reliner, and glaze, in their microbiological, microstructural and mechanical characteristics. A solution containing silica nanoparticles coated by silver nanoparticles in two proportions, 10 millimols and 30 millimols, where they were characterized, was prepared by the hydrolysis and controlled condensation method (Stober method). Silica nanoparticles incorporated with silver were analyzed by integrated X-ray dispersive energy (EDS), scanning electron microscopy (SEM), and dynamic light scattering (DLS).For the microbiological analysis, 10 cylindrical samples (2mmx10mm) were made in chemically activated acrylic resin where the particles were incorporated in two concentrations: 2.5% and 5% with two different molarities of 10 mM and 30 mM. In another situation, 10 cylindrical samples (2mmx10mm) of soft reliner were made, with concentrations of 2.5% and 5% with the two different molarities 10 mM and 30 mM, in the third case the particles were added to a glaze in the two concentrations and molarities and applied under a sample of soft reliner Soft Confort (Dencril). A suspension of Candida albicans and Streptococcus mutans was used for analysis of minimal inhibitory concentration. The samples were made in acrylic resin (n = 6) of 30x10x3mm and were made to perform the three-point flexural strength test of EMIC (Model DL-1000, EMIC Equipamentos e Sistemas Ltda., São José dos Pinhais - PR - Brazil), with a speed of 1mm / min. Samples without the presence of nanoparticles were made as a control group. The silica initially presented as a fine white powder after its characterization with the Silver Nitrate particles, there was a change of coloration to a yellowish tone, increase of its density and the size of its nanoparticles. The results of antimicrobial action were positive for the soft reliner and glaze samples with 5% and 30 mMol of nanoparticles, whereas in the mechanical assay there was no statistically significant difference between the groups (AU)
Assuntos
Humanos , Antifúngicos/imunologia , Resinas Acrílicas/administração & dosagem , Anti-Infecciosos/imunologia , Nanopartículas Metálicas/classificaçãoRESUMO
Melanocytes are dendritic cells located in the skin and mucosae that synthesize melanin. Some infections induce hypo- or hyperpigmentation, which is associated with the activation of Toll-like receptors (TLRs), especially TLR4. Candida albicans is an opportunist pathogen that can switch between blastoconidia and hyphae forms; the latter is associated with invasion. Our objectives in this study were to ascertain whether C. albicans induces pigmentation in melanocytes and whether this process is dependent on TLR activation, as well as relating this with the antifungal activity of melanin as a first line of innate immunity against fungal infections. Normal human melanocytes were stimulated with C. albicans supernatants or with crude extracts of the blastoconidia or hyphae forms, and pigmentation and TLR2/TLR4 expression were measured. Expression of the melanosomal antigens Melan-A and gp100 was examined for any correlation with increased melanin levels or antifungal activity in melanocyte lysates. Melanosomal antigens were induced earlier than cell pigmentation, and hyphae induced stronger melanization than blastoconidia. Notably, when melanocytes were stimulated with crude extracts of C. albicans, the cell surface expression of TLR2/TLR4 began at 48 h post-stimulation and peaked at 72 h. At this time, blastoconidia induced both TLR2 and TLR4 expression, whereas hyphae only induced TLR4 expression. Taken together, these results suggest that melanocytes play a key role in innate immune responses against C. albicans infections by recognizing pathogenic forms of C. albicans via TLR4, resulting in increased melanin content and inhibition of infection.
Assuntos
Candida albicans/patogenicidade , Candidíase/imunologia , Melaninas/metabolismo , Melanócitos/imunologia , Receptor 4 Toll-Like/imunologia , Anticorpos Antifúngicos/imunologia , Antifúngicos/imunologia , Antifúngicos/metabolismo , Antígenos de Fungos/imunologia , Antígenos de Fungos/metabolismo , Candida albicans/imunologia , Candida albicans/metabolismo , Candidíase/microbiologia , Interações Hospedeiro-Patógeno , Humanos , Hifas/fisiologia , Imunidade Inata , Melaninas/imunologia , Melanócitos/metabolismo , Melanócitos/microbiologia , Melanossomas/imunologia , Esporos Fúngicos/fisiologia , Receptor 2 Toll-Like/imunologia , Receptor 2 Toll-Like/metabolismo , Receptor 4 Toll-Like/metabolismoRESUMO
In this study we investigated the effect of beta-glucan derived from Saccharomyces cerevisiae on fungicidal activity, cytokine production and natural killer activity. Spleen and peritoneal cells from female C57BL/6 mice, previously injected (24 or 48 h) with 20 or 100 microg of glucan by i.p. route, were assayed. In vivo beta-glucan administration primed spleen cells for a higher production of IL-12 and TNF-alpha when S. aureus was used as a stimulus. In addition, beta-glucan increased NK spleen cells activity against YAC target cells. Some immunomodulatory activities not yet described for beta-glucan were observed in this work.
Assuntos
Citocinas/imunologia , Células Matadoras Naturais/efeitos dos fármacos , Células Matadoras Naturais/imunologia , Paracoccidioides/efeitos dos fármacos , Saccharomyces cerevisiae/metabolismo , beta-Glucanas/administração & dosagem , beta-Glucanas/imunologia , Animais , Antifúngicos/administração & dosagem , Antifúngicos/imunologia , Relação Dose-Resposta a Droga , Feminino , Fatores Imunológicos/administração & dosagem , Fatores Imunológicos/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Paracoccidioides/citologiaRESUMO
Se ha evaluado la utilidad del metodo de difusion en agar NeoSensitabs para determinar la sensibilidad in vitro de 52 aislamientos de hongos filamentosos dematiaceos a diez antifungicos: anfotericina B.5-fluorocitosina, Ketoconasol, fluconazol, itraconazol, terbinafina, bifonazol, miconazol, clotrimazol, y griseofulvina. Para la preparacion del inoculo se ultilizo un metodo espectrofotometrico empleandose los medios de Shadomy y Casitone agar (CAS), simultaneamente. Todos los aislamientos fueron sensibles, al itraconazol, terbinafina y bifonazol. Al ketonazol el 90,4% resultaron sensibles al miconazol el 71% y al clotrimazol el 46%. El 63% de las cepas fueron sensibles a la anfotericina B y el 28,8% resistente. Por el contrario, el 94,2% de los aislamientos resultaron resistentes a la griseofulvina y el 96% al flucanazol. El 100% de las cepas fueron resistentes a la 5-fluorocitosina. Las zonas de inhibicion no mostraron variaciones en cuanto a la sensibilidad dependiendo del medio; sin embargo, hubo un mejor desarrollo fungico en el medio CAS. Las variaciones en la sensibiblidad observadas con especies como Exophiala spinifera y Fonsecaea pedrosi justificarian el estudio de la sensibiblidad in vitro para valorar el tratamiento clinico con antifungicos. Estos resultados demuestran que el metodo de difusion en agar NeoSensitabs es facil de realizar, rapido y economico por lo que esta al alcance de muchos laboratorios clinicos para el estudio de la sensibilidad in vitro en mochos dematiaceos