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1.
Med Mycol ; 62(9)2024 Sep 06.
Artículo en Inglés | MEDLINE | ID: mdl-39237447

RESUMEN

The immunopathogenesis of recurrent vulvovaginal candidiasis (RVVC) is poorly understood. Recently, it was reported that patients with RVVC present a decrease in both the fungicidal capacity of neutrophils and the proliferative capability of peripheral blood mononuclear cells in response to Candida albicans infection, suggesting an alteration in the innate and adaptive immune response. The aim of this study was to determine the in-situ expression, in the vaginal mucosa, of genes associated with the immune response, as well as the serum concentrations of dectin-1, mannose-binding lectin (MBL), and vitamin D in patients with RVVC. A study was carried out on 40 patients with a diagnosis of RVVC and 26 healthy women. Vaginal scrapings were obtained, and the expression of genes that encode cytokines and transcription factors specific for Th1, Th2, Th17, Treg, pro-inflammatory profiles, and enzymes related to oxidative/microbicidal mechanisms was evaluated by quantitiative polymerase chain reaction (qPCR). Additionally, serum levels of vitamin D and the soluble receptors dectin-1 and MBL were determined by enzyme-linked immunosorbent assay (ELISA). In patients with RVVC, a decreased expression of T-bet, RORγ-T, IL-1ß, and IL-17, and an increase in the expression of FOXP3, IL-4, IL-8, IL-10, and IL-18 were observed when compared to healthy women: moreover, decreased levels of MBL were also observed in these patients. These results confirm that patients with RVVC present in-situ alterations in both the specific and adaptive immune response against Candida spp., a fact that could be associated with the exaggerated vaginal inflammatory response.


The study concerns the immune response of women with recurrent vulvovaginal candidiasis; we observed an alteration in the expression of genes that participate in the control of infection, a fact that could be associated with the exaggerated vaginal inflammatory response observed in those patients.


Asunto(s)
Candidiasis Vulvovaginal , Citocinas , Lectinas Tipo C , Vagina , Vitamina D , Humanos , Candidiasis Vulvovaginal/inmunología , Candidiasis Vulvovaginal/microbiología , Femenino , Lectinas Tipo C/genética , Adulto , Citocinas/sangre , Vagina/microbiología , Vagina/inmunología , Vitamina D/sangre , Adulto Joven , Recurrencia , Lectina de Unión a Manosa/sangre , Lectina de Unión a Manosa/genética , Membrana Mucosa/inmunología , Membrana Mucosa/microbiología , Candida albicans/inmunología
2.
mSphere ; 9(8): e0046724, 2024 Aug 28.
Artículo en Inglés | MEDLINE | ID: mdl-39037263

RESUMEN

Systemic candidiasis remains a significant public health concern worldwide, with high mortality rates despite available antifungal drugs. Drug-resistant strains add to the urgency for alternative therapies. In this context, vaccination has reemerged as a prominent immune-based strategy. Extracellular vesicles (EVs), nanosized lipid bilayer particles, carry a diverse array of native fungal antigens, including proteins, nucleic acids, lipids, and glycans. Previous studies from our laboratory demonstrated that Candida albicans EVs triggered the innate immune response, activating bone marrow-derived dendritic cells (BMDCs) and potentially acting as a bridge between innate and adaptive immunity. Vaccination with C. albicans EVs induced the production of specific antibodies, modulated cytokine production, and provided protection in immunosuppressed mice infected with lethal C. albicans inoculum. To elucidate the mechanisms underlying EV-induced immune activation, our study investigated pathogen-associated molecular patterns (PAMPs) and pattern recognition receptors (PRRs) involved in EVs-phagocyte engagement. EVs from wild-type and mutant C. albicans strains with truncated mannoproteins were compared for their ability to stimulate BMDCs. Our findings revealed that EV decoration with O- and N-linked mannans and the presence of ß-1,3-glucans and chitin oligomers may modulate the activation of specific PRRs, in particular Toll-like receptor 4 (TLR4) and dectin-1. The protective effect of vaccination with wild-type EVs was found to be dependent on TLR4. These results suggest that fungal EVs can be harnessed in vaccine formulations to selectively activate PRRs in phagocytes, offering potential avenues for combating or preventing candidiasis.IMPORTANCESystemic candidiasis is a serious global health concern with high mortality rates and growing drug resistance. Vaccination offers a promising solution. A unique approach involves using tiny lipid-coated particles called extracellular vesicles (EVs), which carry various fungal components. Previous studies found that Candida albicans EVs activate the immune response and may bridge the gap between innate and adaptive immunity. To understand this better, we investigated how these EVs activate immune cells. We demonstrated that specific components on EV surfaces, such as mannans and glucans, interact with receptors on immune cells, including Toll-like receptor 4 (TLR4) and dectin-1. Moreover, vaccinating with these EVs led to strong immune responses and full protection in mice infected with Candida. This work shows how harnessing fungal EVs might lead to effective vaccines against candidiasis.


Asunto(s)
Candida albicans , Candidiasis , Células Dendríticas , Vesículas Extracelulares , Vacunas Fúngicas , Receptores de Reconocimiento de Patrones , Receptor Toll-Like 4 , Animales , Candida albicans/inmunología , Vesículas Extracelulares/inmunología , Receptor Toll-Like 4/inmunología , Receptor Toll-Like 4/metabolismo , Ratones , Candidiasis/inmunología , Candidiasis/prevención & control , Candidiasis/microbiología , Vacunas Fúngicas/inmunología , Vacunas Fúngicas/administración & dosificación , Células Dendríticas/inmunología , Receptores de Reconocimiento de Patrones/inmunología , Ratones Endogámicos C57BL , Femenino , Inmunidad Innata , Modelos Animales de Enfermedad
3.
Rev Assoc Med Bras (1992) ; 70(4): e20230845, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38747876

RESUMEN

OBJECTIVE: COVID-19 infection poses significant risks, including life-threatening consequences and fungus synchronization, making it a significant concern. This study seeks to assess the effect of concurrent infection of COVID-19 with Thrush Candida albicans on the patient's health state by measuring the proportion of immune cells and certain interleukins such as IL-8, -10, -17, and -33. METHODS: The study involved 70 patients (30 patients with COVID-19, 17 patients with thrush candidiasis, and 23 patients with Thrush Candida albicans) and 50 healthy individuals as a control group. COVID-19 was identified using RT-PCR, while C. albicans were identified through culture media, biochemical testing, and oral swabs. Ruby equipment and ELISA kits were used for blood counts and interleukin detection. RESULTS: COVID-19, thrush candidiasis, and Thrush Candida albicans infections occur in a wide range of age groups (4-80 years), with no significant differences between sexes (p>0.05). Immunologically, our study found that Thrush Candida albicans patients had the highest rate of neutrophils (89.6%) and basophils (2.01%), while corona patients had the highest percentage of lymphocytes (70.12%) and eosinophils (7.11%), and patients with thrush candidiasis had the highest percentage of monocytes. Thrush Candida albicans patients showed increased IL-8 (56.7 pg/mL) and IL-17 (101.1 pg/mL) concentrations, with the greatest concentration of IL-33 (200.5 pg/mL) in COVID-19, and a decrease in the level of IL-10 in patient groups compared with controls. CONCLUSION: Patient groups showed increased neutrophils, lymphocytes, monocytes, and IL-8 levels, with a significant linear association between proinflammatory interleukins and these cells.


Asunto(s)
Biomarcadores , COVID-19 , Humanos , COVID-19/inmunología , COVID-19/complicaciones , Masculino , Femenino , Persona de Mediana Edad , Adulto , Biomarcadores/sangre , Biomarcadores/análisis , Anciano , Adolescente , Adulto Joven , Estudios de Casos y Controles , Coinfección/inmunología , Coinfección/sangre , SARS-CoV-2/inmunología , Candidiasis Bucal/inmunología , Interleucinas/sangre , Anciano de 80 o más Años , Candida albicans/aislamiento & purificación , Candida albicans/inmunología , Niño , Preescolar
4.
Future Microbiol ; 19(7): 585-594, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38629904

RESUMEN

Aim: To evaluate the behavior of oral keratinocytes in the presence of Vitamin C (Vit C) and its anti-inflammatory potential. Materials & methods: Oral keratinocytes were initially exposed to 0.1-2.5 mM of Vit C and the metabolic activity and cell migration were evaluated using MTS assay and Ibidi culture inserts, respectively. After, the cells were challenged with Candida albicans and inflammatory markers were analyzed by qPCR. Results: The treatment was not cytotoxic, and the highest concentrations increased the metabolic activity at 24 h. Vit C delayed the cell migration at 48 and 72 h. Interestingly, it downregulated the genes IL-8 and IL-1ß. Conclusion: Vit C could be an interesting adjuvant to anti-fungal treatment due to its anti-inflammatory potential.


Vitamin C, also known as ascorbic acid, is a vitamin commonly found in fruits and vegetables. It is popular for supporting our immune system, so is commonly taken as a supplement. We looked at the action of vitamin C on cells from the mouth and its potential to reduce inflammation in a fungal disease of the mouth ­ oral candidiasis. We showed that vitamin C is not toxic to cells of the mouth and may reduce inflammation in cells infected by the fungus. This suggests that vitamin C could be used as a complementary therapy for oral candidiasis.


Asunto(s)
Antiinflamatorios , Ácido Ascórbico , Candida albicans , Movimiento Celular , Queratinocitos , Candida albicans/efectos de los fármacos , Candida albicans/inmunología , Ácido Ascórbico/farmacología , Humanos , Queratinocitos/efectos de los fármacos , Queratinocitos/inmunología , Queratinocitos/microbiología , Queratinocitos/metabolismo , Antiinflamatorios/farmacología , Movimiento Celular/efectos de los fármacos , Interleucina-8/metabolismo , Interleucina-8/genética , Interleucina-1beta/metabolismo , Interleucina-1beta/genética , Inflamación , Antifúngicos/farmacología
5.
Sci Rep ; 11(1): 20281, 2021 10 13.
Artículo en Inglés | MEDLINE | ID: mdl-34645905

RESUMEN

Fungal infections represent a major global health problem affecting over a billion people that kills more than 1.5 million annually. In this study, we employed an integrative approach to reveal the landscape of the human immune responses to Candida spp. through meta-analysis of microarray, bulk, and single-cell RNA sequencing (scRNA-seq) data for the blood transcriptome. We identified across these different studies a consistent interconnected network interplay of signaling molecules involved in both Toll-like receptor (TLR) and interferon (IFN) signaling cascades that is activated in response to different Candida species (C. albicans, C. auris, C. glabrata, C. parapsilosis, and C. tropicalis). Among these molecules are several types I IFN, indicating an overlap with antiviral immune responses. scRNA-seq data confirmed that genes commonly identified by the three transcriptomic methods show cell type-specific expression patterns in various innate and adaptive immune cells. These findings shed new light on the anti-Candida immune response, providing putative molecular pathways for therapeutic intervention.


Asunto(s)
Candida albicans/inmunología , Candida glabrata/inmunología , Candida parapsilosis/inmunología , Candidiasis/inmunología , Candidiasis/microbiología , Transducción de Señal/inmunología , Antivirales/farmacología , Biología Computacional/métodos , Bases de Datos Genéticas , Perfilación de la Expresión Génica , Regulación Fúngica de la Expresión Génica , Humanos , Inmunidad , Inmunidad Innata , Interferones/metabolismo , RNA-Seq , Transcripción Genética , Transcriptoma
6.
Biomolecules ; 11(8)2021 08 13.
Artículo en Inglés | MEDLINE | ID: mdl-34439875

RESUMEN

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.


Asunto(s)
Antifúngicos/inmunología , Candida albicans , Candidiasis , Farmacorresistencia Fúngica , Histatinas/inmunología , Saliva/inmunología , Animales , Candida albicans/inmunología , Candida albicans/fisiología , Candidiasis/tratamiento farmacológico , Candidiasis/inmunología , Humanos
7.
Immunol Res ; 69(3): 264-274, 2021 06.
Artículo en Inglés | MEDLINE | ID: mdl-33966252

RESUMEN

Neutrophil extracellular traps (NETs) are networks of decondensed chromatin loaded with antimicrobial peptides and enzymes produced against microorganisms or biochemical stimuli. Since their discovery, numerous studies made separately have revealed multiple triggers that induce similar NET morphologies allowing to classify them as lytic or non-lytic. However, the variability in NET composition depending on the inducer agent and the local milieu under similar conditions has been scarcely studied. In this work, a comparative study was conducted to evaluate structural and enzymatic divergences in NET composition induced by biochemical (phorbol myristate acetate [PMA] and hypochlorous acid [HOCl]) and microbiologic (Candida albicans, Staphylococcus aureus, and Pseudomonas aeruginosa) stimuli, along with the presence of plasma from healthy donors or patients with systemic lupus erythematosus (SLE). The results showed a differential composition of DNA and the antimicrobial peptide cathelicidin (LL37) and a variable enzymatic activity (neutrophil elastase, cathepsin G, myeloperoxidase) induced by the different stimuli despite showing morphologically similar NETs. Additionally, SLE plasma´s presence increased DNA and LL37 release during NET induction independently of the trigger stimulus but with no enzymatic activity differences. This work provides new evidence about NET composition variability depending on the inducer stimulus and the local milieu.


Asunto(s)
Trampas Extracelulares/metabolismo , Lupus Eritematoso Sistémico/inmunología , Neutrófilos/metabolismo , Biomarcadores/análisis , Biomarcadores/metabolismo , Candida albicans/inmunología , Estudios de Casos y Controles , Catelicidinas/análisis , Catelicidinas/metabolismo , Catepsina G/análisis , Catepsina G/metabolismo , Células Cultivadas , Trampas Extracelulares/inmunología , Voluntarios Sanos , Humanos , Ácido Hipocloroso/inmunología , Elastasa de Leucocito/análisis , Elastasa de Leucocito/metabolismo , Lupus Eritematoso Sistémico/sangre , Neutrófilos/inmunología , Peroxidasa/análisis , Peroxidasa/metabolismo , Cultivo Primario de Células , Pseudomonas aeruginosa/inmunología , Staphylococcus aureus/inmunología , Acetato de Tetradecanoilforbol/inmunología
8.
São José dos Campos; s.n; 2021. 53 p. tab.il., graf..
Tesis en Portugués | BBO - Odontología | ID: biblio-1281107

RESUMEN

Candida albicans é um fungo que habitualmente coloniza superfícies mucosas de humanos e pode assumir caráter patogênico a depender de fatores do hospedeiro. Portadores da Síndrome da Imunodeficiência Humana, causada pelo Vírus da Imunodeficiência Humana (HIV), são propícios a apresentar candidose nas mucosas devido a imunodeficiência celular que apresentam. A introdução da Terapia Antirretroviral (TARV), em especial o surgimento dos Inibidores da Protease do HIV (IPs-HIV), reduziu drasticamente a incidência e prevalência destas patologias ao longo dos anos. Estudos clínicos e epidemiológicos com IPs-HIV de primeira geração demostraram que tal redução não se deve exclusivamente à melhora imunológica promovida pela TARV, e pesquisas in vitro já demonstraram propriedades antifúngicas e antibiofilme de alguns IPs-HIV de primeiras gerações em C. albicans. Sendo assim, o objetivo deste estudo foi avaliar os efeitos do Atazanavir (ATV) e Darunavir (DRV), dois IPs-HIV de segunda e terceira geração respectivamente em uso clínico atual no Brasil, em diferentes fatores de virulência de C. albicans. Para isso foram realizados estudos com duas cepas clínicas de C. albicans isoladas de lesões de candidose orofaríngea de pacientes portadores de HIV para avaliar a ação in vitro das drogas na morfogênese, formação de biofilme (contagem de células viáveis e quantificação de biomassa) e na expressão dos genes de virulência BRC1 e SAP2, e in vivo no efeito protetor desses medicamentos na infecção experimental por C. albicans em modelo de Galleria mellonella. Os dados foram analisados por teste t, ANOVA, Kruskal-Wallis, Dunn e Kaplan-Meier (p<0,05). A Concentração Inibitória Mínima (CIM) para ambos os IPs-HIV testados foi 512 µg/mL. Nos biofilmes, a redução na contagem de UFC/mL de C. albicans nos grupos tratados com IPs-HIV foi de até 6,81 Log. A biomassa dos biofilmes tratados também sofreu reduções significantes para ATV (82%), DRV (81%) comparada ao grupo controle.DRV e ATV promoveram redução estatisticamente significante de expressão gênica de SAP2 e BRC1, respectivamente, quando comparados ao controle (p<0,05). Em relação à morfogênese de C. albicans, ATV e DRV inibiram significativamente a formação de hifas (p=0,0183). No estudo in vivo, o uso profilático de ATV e DRV em G. mellonella infectadas com C. albicans prolongou em até 40% a sobrevivência das larvas (p=0,0004). Conclui-se que ATV e DRV inibiram a filamentação e apresentaram atividade antifúngica, antibiofilme e na expressão de genes de fatores de virulência de C. albicans e preveniram candidose em G. mellonella(AU)


Candida albicans is a fungus that usually colonizes mucous surfaces of humans and can assume pathogenic character depending on host factors. Patients with Human Immunodeficiency Syndrome, caused by the Human Immunodeficiency Virus (HIV), are favorable to present mucous candidosis due to the cellular immunodeficiency they present. The introduction of Antiretroviral Therapy (ART), especially the emergence of HIV Protease Inhibitors (HIV-PI), has drastically reduced the incidence and prevalence of these mycosis over the years. Clinical and epidemiological studies with firstgeneration HIV-PIs have shown that this reduction is not exclusively due to the immunological improvement promoted by ART, and in vitro research has already demonstrated antifungal and antibiofilm properties of firsts-generations HIV-PIs on C. albicans. Thus, the aim of this study was to evaluate the effects of Atazanavir (ATV) and Darunavir (DRV), two HIV-PIs of second and third generation respectively currently in clinical use in Brazil, on virulence factors of C. albicans. For this purpose, studies were carried out with two clinical strains of C. albicans isolated from lesions of oropharyngeal candidosis of HIV positive patients to evaluate in vitro action of the drugs on morphogenesis, biofilm formation (number of viable cell and biomass determination) and virulence factor genes BRC1 and SAP2 expression and in vivo on the protective effect of these drugs on experimental infection by C. albicans in Galleria mellonella model. Data were analyzed by t Student, ANOVA, Kruskal-Wallis and Dunn and Kaplan-Meier (p<0.05) tests. The Minimum Inhibitory Concentration (MIC) for both tested HIV-PIs was 512 µg/mL. In biofilms, a reduction in the CFU/mL count of C. albicans in the groups treated with HIV-PIs was up to 6.81 log. The biomass of biofilms also suffered significant reductions for ATV (82%) and DRV (81%) compared to the control group. DRV and ATV statistically significantly downregulated the expression of SAP2 and BRC1 genes respectively (p<0,05). Regarding the morphogenesis of C. albicans, ATV and DRV inhibited the formation of hyphae (p = 0.0183). In the in vivo study, the prophylactic use of ATV and DRV in G. mellonella infected with C. albicans prolonged larval survival by up to 40% (p = 0.0004). We can conclude that ATV and DRV inhibited filamentation and showed antifungal activity, being able to inhibit growth, formation and virulence factors gene expression of C. albicans biofilm and prevented candidosis in G. mellonel(AU)


Asunto(s)
Candida albicans/inmunología , Inhibidores de la Proteasa del VIH/administración & dosificación , Factores de Virulencia/efectos adversos , Darunavir/síntesis química , Sulfato de Atazanavir/efectos adversos
9.
Rev Alerg Mex ; 67(2): 119-127, 2020.
Artículo en Español | MEDLINE | ID: mdl-32892527

RESUMEN

BACKGROUND: The prevalence of allergic diseases has increased worldwide. Recent studies have informed that the dysbiosis of some specific members of the human microbiota may enhance the allergic response of the respiratory tract. OBJECTIVE: To retrospectively explore the role of some microorganisms of the human microbiota on the skin reactivity and their effect on the chronicity of allergic respiratory diseases in humans. METHODS: A retrospective analysis of a 5-year database of patients with allergic respiratory tract disease. The frequency and magnitude of the reactivity to 38 different allergens was determined. RESULTS: Dermatophagoides pteronyssinus had the highest frequency of reactivity (93.7 %), followed by the bacterial allergen (a mixture of Staphylococcus aureus and Staphylococcus epidermidis) with a frequency of reactivity of 91.82 %; whereas Candida albicans had a frequency of reactivity of only 79.32 %. The frequency of reactivity to the pollen of native Mexican weeds was even lower ~79 %. CONCLUSION: The microorganisms of the microbiota that were analyzed in this study seem to have an influence on the development of respiratory allergic inflammation, associated with long-term colonization of the pharynx, nasal mucosa, and sinuses because of these microorganisms.


Antecedentes: La prevalencia de las enfermedades alérgicas ha aumentado en todo el mundo. En estudios recientes se ha informado que la disbiosis de algunos miembros específicos de la microbiota humana puede potenciar la respuesta alérgica de las vías respiratorias. Objetivo: Explorar retrospectivamente el papel de algunos microorganismos de la microbiota humana en la reactividad cutánea y su efecto sobre la cronicidad de las enfermedades alérgicas respiratorias en el humano. Métodos: Análisis retrospectivo de la base de datos de un periodo de cinco años de pacientes con enfermedad alérgica de las vías respiratorias. Se determinó la frecuencia y magnitud de la reactividad a 38 alérgenos diferentes. Resultados: La mayor frecuencia de reactividad la presentó Dermatophagoides pteronyssinus (93.7 %), al que le siguió una combinación bacteriana de Staphylococcus aureus-Staphylococcus epidermidis (91.82 %) y Candida albicans (79.32 %). La reactividad a alérgenos de polen de malezas nativas de México fue aun menor, aproximadamente de 79 %. Conclusión: Los microorganismos de la microbiota analizados en este estudio parecen tener una influencia en el desarrollo de la inflamación alérgica respiratoria, asociada a la colonización a largo plazo de la faringe, la mucosa nasal y los senos paranasales.


Asunto(s)
Alérgenos/inmunología , Antígenos/inmunología , Microbiota/inmunología , Hipersensibilidad Respiratoria/inmunología , Sistema Respiratorio/inmunología , Animales , Antígenos Bacterianos/inmunología , Candida albicans/inmunología , Niño , Dermatophagoides pteronyssinus/inmunología , Femenino , Humanos , Masculino , Estudios Retrospectivos , Staphylococcus aureus/inmunología , Staphylococcus epidermidis/inmunología , Adulto Joven
10.
BMC Microbiol ; 20(1): 245, 2020 08 06.
Artículo en Inglés | MEDLINE | ID: mdl-32762645

RESUMEN

BACKGROUND: Peptidorhamnomannan is a glycoconjugate that consists of a peptide chain substituted by O- and N-linked glycans, present on the cell surface of Lomentospora prolificans, a saprophytic fungus which is widely distributed in regions with temperate climates. O-linked oligosaccharides from peptidorhamnomannan isolated from Lomentospora prolificans conidia are recognized by macrophages mediating macrophage - conidia interaction. In this work, peptidorhamnomannan was isolated from L. prolificans mycelium cell wall and its role in macrophage - Candida albicans interaction was evaluated. RESULTS: Purified peptidorhamnomannan inhibits the reactivity of rabbit immune sera to mycelial and conidia forms of L. prolificans, indicating that this glycoconjugate is exposed on the fungal surface and can mediate interaction with host immune cells. We demonstrated that peptidorhamnomannan leads to TNF-α production in J774 macrophages for 1, 2 and 3 h of incubation, suggesting that this glycoconjugate may have a beneficial role in the response to fungal infections. In order to confirm this possibility, the effect of peptidorhamnomannan on the macrophage - C. albicans interaction was evaluated. Macrophages treated with peptidorhamnomannan led to a lower fungal survival, suggesting that peptidorhamnomannan induces an increased fungicidal activity in macrophages. Furthermore, TNF-α levels were measured in supernatants after macrophage - C. albicans interaction for 1, 2 and 3 h. Peptidorhamnomannan treatment led to a higher TNF-α production at the beginning of the interaction. However, the release of TNF-α was not maintained after 1 h of incubation. Besides, peptidorhamnomannan did not show any inhibitory or fungicidal effect in C. albicans when used at 100 µg/ml but it was able to kill C. albicans at a concentration of 400 µg/ml. CONCLUSION: We suggest that peptidorhamnomannan acts as a molecular pattern on the invading pathogen, promotes TNF-α production and, thus, increases macrophage fungicidal activity against Candida albicans.


Asunto(s)
Candida albicans/inmunología , Glicoproteínas/farmacología , Macrófagos/citología , Scedosporium/metabolismo , Animales , Candida albicans/patogenicidad , Línea Celular , Pared Celular/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Sueros Inmunes/efectos de los fármacos , Sueros Inmunes/metabolismo , Macrófagos/efectos de los fármacos , Macrófagos/inmunología , Macrófagos/microbiología , Ratones , Micelio/metabolismo , Fagocitosis , Conejos , Factor de Necrosis Tumoral alfa/metabolismo
11.
Cell Microbiol ; 22(10): e13238, 2020 10.
Artículo en Inglés | MEDLINE | ID: mdl-32558196

RESUMEN

Extracellular vesicles (EVs) are lipid bilayered compartments released by virtually all living cells, including fungi. Among the diverse molecules carried by fungal EVs, a number of immunogens, virulence factors and regulators have been characterised. Within EVs, these components could potentially impact disease outcomes by interacting with the host. From this perspective, we previously demonstrated that EVs from Candida albicans could be taken up by and activate macrophages and dendritic cells to produce cytokines and express costimulatory molecules. Moreover, pre-treatment of Galleria mellonella larvae with fungal EVs protected the insects against a subsequent lethal infection with C. albicans yeasts. These data indicate that C. albicans EVs are multi-antigenic compartments that activate the innate immune system and could be exploited as vaccine formulations. Here, we investigated whether immunisation with C. albicans EVs induces a protective effect against murine candidiasis in immunosuppressed mice. Total and fungal antigen-specific serum IgG antibodies increased by 21 days after immunisation, confirming the efficacy of the protocol. Vaccination decreased fungal burden in the liver, spleen and kidney of mice challenged with C. albicans. Splenic levels of cytokines indicated a lower inflammatory response in mice immunised with EVs when compared with EVs + Freund's adjuvant (ADJ). Higher levels of IL-12p70, TNFα and IFNγ were detected in mice vaccinated with EVs + ADJ, while IL-12p70, TGFß, IL-4 and IL-10 were increased when no adjuvants were added. Full protection of lethally challenged mice was observed when EVs were administered, regardless the presence of adjuvant. Physical properties of the EVs were also investigated and EVs produced by C. albicans were relatively stable after storage at 4, -20 or -80°C, keeping their ability to activate dendritic cells and to protect G. mellonella against a lethal candidiasis. Our data suggest that fungal EVs could be a safe source of antigens to be exploited in vaccine formulations.


Asunto(s)
Candida albicans/inmunología , Candidiasis/inmunología , Vesículas Extracelulares/inmunología , Animales , Anticuerpos Antifúngicos/sangre , Antígenos Fúngicos/inmunología , Candidiasis/prevención & control , Frío , Citocinas/sangre , Células Dendríticas/inmunología , Células Dendríticas/metabolismo , Femenino , Vacunas Fúngicas/inmunología , Inmunoglobulina G/sangre , Inmunoglobulina M/sangre , Interleucina-6/biosíntesis , Ratones , Ratones Endogámicos BALB C , Mariposas Nocturnas/inmunología , Mariposas Nocturnas/microbiología , Vacunación
13.
São José dos Campos; s.n; 2020. 49 p. il., graf., tab..
Tesis en Portugués | BBO - Odontología | ID: biblio-1150738

RESUMEN

A crescente incidência da resistência às drogas antifúngicas tornou-se um grande desafio para áreas médica e odontológica, fazendo com que a busca por métodos de tratamento alternativos ou em combinação com as já utilizadas sejam urgentemente necessárias. O objetivo deste projeto foi avaliar os efeitos da associação entre o plasma frio em pressão atmosférica e antifúngicos poliênicos convencionais utilizados no tratamento tópico da candidose bucal. Para tanto, foram realizados: a) Determinação das concentrações inibitórias dos antifúngicos poliênicos nistatina e anfotericina B e do plasma frio em pressão atmosférica sobre biofilmes de C. albicans; b) Avaliação do efeito dos tratamentos isolados e associados em condições/concentrações sub-inibitórias para fins de comparação e c) Avaliação dos diferentes protocolos de aplicação dos tratamentos, visando a obtenção da condição experimental mais eficaz para o controle do biofilme fúngico. Os dados de unidades formadoras de colônia foram comparados estatisticamente entre os grupos por One-way ANOVA e post hoc Tukey, com nível de significância de 5%. A partir dos resultados obtidos, pode-se concluir que o jato de plasma apresentou efeito antifúngico similar quando aplicado isoladamente ou em associação com nistatina e anfotericina B. Ainda, foi possível concluir que o jato de plasma frio apresenta efeito antifúngico mais eficaz do que os tratamentos dos biofilmes utilizando nistatina e anfotericina B no tempo de 5 minutos(AU)


The increasing incidence of antifungal resistance represents a great challenge in the medical areas and Dentistry. For this reason, the search for alternative methods or combination with the conventional ones is urgently necessary. The aim of this project is to evaluate, the effects of association between atmospheric pressure cold plasma and conventional polyene antifungals used for the treatment of oral candidiasis will be studied. With this purpose, the following methodologies were be used: a) To determine the inhibitory concentrations of the polyene antifungals nystatin and amphotericin B and cold plasma on C. albicans biofilms, b) To evaluate the combination of treatments in subinhibitory conditions, including isolated treatments for comparison purposes and c) To evaluate different protocols of treatment application, aiming to obtain the most effective protocol against fungal biofilm. Data was compared by One-way ANOVA and post hoc Tukey, with a significance level of 5%. Considering the results, it could be concluded that cold plasma showed similar effects when applied alone or in association to nystatin and amphotericin B. Cold plasma showed more effective antifungal effect on biofilms when compared to nystatin and amphotericin B, after 5 min exposure(AU)


Asunto(s)
Candida albicans/inmunología , Plasma/enzimología , Antifúngicos/farmacología
14.
São Jose dos Campos; s.n; 2020. 90 p. il., graf., tab..
Tesis en Portugués | LILACS, BBO - Odontología | ID: biblio-1223429

RESUMEN

O objetivo desse estudo foi sintetizar um scaffold de nanofibras (NFs) de Policaprolactona (PCL) com adição de nistatina (NYS), caracterizar e avaliar a atividade antimicrobiana. NFs foram sintetizadas pela técnica da eletrofiação, utilizando-se solução de PCL puro e PCL adicionado de NYS. Para a síntese das NFs de PCL, o polímero foi dissolvido em Dimetilformamida (DMF) e 1,1,2,2 Tetracloroetano (TCE), logo após foram adicionadas três concentrações de NYS (Grupos A, B e C). As amostras foram analisadas em Microscopia Eletrônica de Varredura (MEV), Espectroscopia por energia dispersiva (EDS), Análise de molhabilidade, Análise de Difratometria de Raios-X (DRX) e Espectroscopia de Infravermelho por Transformada de Fourier (FTIR). Após a análise em MEV foi realizada a média dos diâmetros das fibras com Software ImageJ. As atividades antimicrobianas foram avaliadas por meio dos testes de concentração inibitória mínima (CIM) e concentração microbicida mímica (CMM). As NFs de PCL com NYS (A) apresentaram diâmetro de 0,99µm ±0,29, o padrão do grupo B apresentou uma média de diâmetro de 1,3µm ±0,56 e o grupo C 1,24µm ±0,48. O DRX e o EDS comprovaram a presença de PCL e de NYS nas amostras. A CIM e a CMM comprovaram a ação fungicida e fungistática das NFs A, B e C(AU)


The aim of this study was to synthesize a scaffold of policaprolactane (PCL) nanofibers (NFs) with addition of nystatin (NYS), to characterize and evaluate the antimicrobial activity. NFs were synthesized by the electro-spinning technique, using pure PCL and NYS and PCL solutions. For the synthesis of PCL NFs, the polymer was dissolved in Dimethylformamide (DMF) and 1,1,2,2- Tetrachloroethane (TCE), after which three concentrations of NYS (Groups A, B and C) were added. The samples were analyzed in Scanning Electron Microscopy (SEM), Dispersive Energy Spectroscopy (EDS), Goniometer, X-ray Diffraction Analysis (XRD) and Fourier Transform Infrared Spectroscopy (FTIR). After SEM analysis, the mean fiber diameter was performed with ImageJ Software. The antimicrobial activities were evaluated through the tests of minimum inhibitory concentration (MIC) and minimum microbicidal concentration (MMC).The NFs of PCL with NYS (A) had a diameter of 0.99 µm ± 0.29, the standard of group B presented a mean diameter of 1.3 µm ± 0.56 and group C 1.24 µm ± 0.48. The DRX and the EDS verified the presence of PCL and NYS in the samples. CIM and CMM proved the fungicidal and fungistatic action of NFs A, B and C(AU)


Asunto(s)
Candida albicans/inmunología , Electroquímica/métodos
15.
Mycopathologia ; 184(6): 747-757, 2019 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-31637573

RESUMEN

Candida albicans is a commensal fungus of the skin and mucous membranes in humans, but it is also responsible for mucocutaneous and systemic infections in immunocompromised patients like low birth weight neonates and premature newborns. The epicutaneous application of C. albicans is widely used to study the immune response against this pathogen in adult mice models. However, the immune response of newborns against infections caused by the genus Candida is poorly understood. In order to mimic premature human infection, we developed a model of C. albicans epicutaneous infection in newborn mice. We found that yeasts were able to colonize while the pseudohyphae invaded the epidermis. Recruitment of polymorphonuclear and mononuclear cells at the infection zone was observed. Fungal invasion, fungal burden and cellular infiltration displayed a time- and dose-dependent response. Interestingly, newborn mice were able to control C. albicans primary infection. Finally, we showed that the epicutaneous infection of C. albicans in newborn mice at birth results in the induction of cell-mediated immunity as evinced by delayed-type hypersensitivity assays.


Asunto(s)
Animales Recién Nacidos/microbiología , Candida albicans/inmunología , Candidiasis/inmunología , Inmunidad Celular , Animales , Candida albicans/crecimiento & desarrollo , Candidiasis/microbiología , Epidermis/microbiología , Ratones , Modelos Animales , Piel/microbiología
16.
São José dos Campos; s.n; 2019. 65 p. il., tab., graf..
Tesis en Portugués | BBO - Odontología | ID: biblio-1024924

RESUMEN

As infecções causadas por Candida albicans são consideradas um desafio importante na área médica. Além dos antifúngicos convencionais apresentarem elevada toxicidade, casos de resistência de C. albicans a estes medicamentos têm sido descritos com frequência. Devido a estas limitações do uso de antifúngicos, a busca por terapias alternativas é alvo crescente de estudos. Dentre as novas terapias, o jato de plasma frio destaca-se a como agente eficaz frente a cepas de C. albicans, sendo considerada uma alternativa promissora. Seu efeito antifúngico é associado à presença de espécies reativas de oxigênio e nitrogênio liberadas, assim como de íons e fótons. Todavia até o presente momento, o exato mecanismo de ação não foi elucidado. Portanto, o objetivo deste estudo é prospectar os efeitos do jato de plasma frio sobre C. albicans. Perseguindo este objetivo, foram utilizadas duas técnicas complementares às técnicas microbiológicas clássicas, a espectroscopia no Infra-vermelho (FT-IR) e a microscopia de força atômica (AFM). Suspensões padronizadas da cepa C. albicans SC 5314 e 1 cepa clínica foram expostas ao jato de plasma. A seguir, foram analisadas por espectroscopia no infra-vermelho (FT-IR) e microscopia de força atômica (AFM) para identificar os efeitos do plasma sobre a parede celular de C. albicans. Controle negativo (sem exposição) e positivo (caspofungina) também foram analisados. Os ensaios foram realizados em triplicata em três experimentos independentes e a análise estatística foi realizada ao nível de significância de 5 %. As alterações morfológicas e topográficas nas células tratadas com plasma foram similares às do grupo tratado com caspofungina. O perfil bioquímico das células após tratamento com o plasma foi também similar ao das células tratadas com caspofungina. Conclui-se que o plasma agiu diretamente sobre as estruturas da parede celular fúngica, mais especificamente sobre os polissacarídeos, com provável influencia na redução de glucanos e aumento de mananas e quitinas. Esse remodelamento da parede celular pode ter correlação com as alterações morfológicas observadas, tais como mudança no tamanho da célula e aumento da rugosidade superficial(AU)


Infections caused by Candida albicans are considered important challenges in the medical area. Besides the high toxicity of conventional antifungals, cases of C. albicans resistance have been reported with increasing frequency. Due to these limitations of the antifungals, the search for alternative therapies is increasing. Among these therapies, cold plasma showed effectiveness against C. albicans isolates and is considered a promising alternative. Its antifungal effect is associated to the presence of oxygen and nitrogen reactive species, as well as ions. Nonetheless, the exact mechanism of action on C. albicans has not yet been elucidated. The aim of this study is to investigate the effects of plasma jet on C. albicans cell wall. For this purpose, two techniques complementing to classic the microbiological methods were used, the infrared spectroscopy (FT-IR) and atomic force microscopy (AFM). Standardized suspensions of C. albicans SC 5314 and 1 clinical isolate was exposed to plasma jet. Samples were analyzed by FT-IR and AFM to identifying the effects of plasma on C. albicans. Negative control (no exposition) and positive (caspofungin) were also analyzed. The experiments will be done in triplicate in three independent experiments and the statistical analyzes were done adopting the level of significance of 5%. The morphological and topographic alterations in cells treated with plasma were similar to the group treated with caspofungin. The biochemical profile of the cells after treatment with the plasma jet was also similar to those treated with caspofungin. It could be concluded that plasma acted directly on the structures of fungal cell wall, more specifically on polysaccharides, influencing the reduction of glucans and increased of mannans and chitins. This remodeling of cell wall can be correlated with the detected morphological alterations, such as changes in cell size and increase in superficial roughness(AU)


Asunto(s)
Humanos , Candida albicans/inmunología , Espectroscopía Infrarroja por Transformada de Fourier/métodos , Microscopía de Fuerza Atómica/métodos
17.
São José dos Campos; s.n; 2019. 98 p. il., graf., tab.
Tesis en Portugués | BBO - Odontología | ID: biblio-995547

RESUMEN

O aumento crescente da prevalência das infecções fúngicas no cenário mundial e aumento da ocorrência de resistência aos antifúngicos convencionais sugerem que pesquisas para a descoberta de novas moléculas antifúngicas são urgentemente necessárias. O objetivo geral deste estudo é contribuir para a prospecção de alternativas terapêuticas, avaliando nanoencapsulados da substância bioativa ácido elágico (NAE) no tratamento da candidose bucal. Os seguintes objetivos específicos foram: complexar ácido elágico em ciclodextrina, em concentração com atividade antifúngica frente C. albicans; caracterizar quimicamente o encapsulado; prospectar possíveis mecanismos de ação; avaliar ação de concentrações subinibitórias sobre a produção de exoenzimas e aderência às células epiteliais; avaliar citotoxicidade do NAE nas condições de tratamento efetivas in vitro e in vivo (Drosophila melanogaster) e avaliar ação no tratamento in vivo de lesões de candidose oral induzida em modelo murino. Os dados foram analisados estatisticamente, adotando-se significância de 5%. HP-ß-CD formou complexos solúveis de inclusão com AE. A porcentagem de AE/HP-ß-CD foi de 15,25 ± 0,49%. As análises de MEV e FTIR confirmaram a formação de complexo de inclusão. AE/HP-ß-CD manteve a atividade antifúngica do AE puro, com CIM 25 µg/ml para C. albicans ATCC 18804 e 50 µg/ml para C. albicans SC 5314. O tratamento de C. albicans com sub-CIMs indicou aumento da CIM na presença de sorbitol para ATCC 18804 (50 µg/ml), sugerindo ação sobre parede celular. Os resultados sugerem que não há ação sobre membrana celular. Não foi detectado efeito sobre morfogênese e produção de enzimas extracelulares. Efeito protetor de AE a 3,2, 6,4 e 32 µg/ml foi observado em modelo D. melanogaster infectado com C. albicans que resultou na sobrevivência de 45, 33 e 34%. Baixa toxicidade foi observada nas concentrações de 200 e 250 µg/ml. No modelo de candidose bucal em camundongos foi observada redução significativa da invasão de hifas no epitélio no grupo tratado AE/HP-ß-CD em relação aos controles, em 24 horas e 48 horas. Conclui-se que AE/HP-ß-CD apresentou efeito inibitório sobre C. albicans in vitro, com baixa toxicidade. Em modelo Drosophila melanogaster apresentou efeito protetor frente à infecção fúngica. Em modelo murino levou à redução na invasão epitelial pelo fungo(AU)


The increase in the prevalence of fungal infections worldwide and the rise in the occurrence of antifungal resistance suggest that new researches for the discovery of antifungal molecules are needed. The aim of this study is to contribute to the prospection of therapeutic alternatives, evaluating nanoencapsulates of ellagic acid (NAE) for the treatment of oral candidosis. In order to reach the general aim, the following specific objectives were determined: to complex the ellagic acid in cyclodextrin at effective concentration against C. albicans; to chemically characterize the encapsulate; to test the activity of the encapsulate using an invasive candidosis in vitro model, searching for the mechanisms of action; to evaluate the activity of subinhibitory concentrations on the exoenzymes production, and adherence to epithelial cells; to evaluate the cytotoxicity of NAE in the effective treatment conditions in vitro and in vivo (Drosophila melanogaster); and to evaluate the in vivo effectiveness for the treatment of oral candidosis in murine model. The obtained data was statistically analyzed (level of significance 5%). HP-ß-CD formed soluble inclusion complexes with EA. The percentage of EA/HP-ß-CD was 15.25 ± 0.49%. SEM and FTIR analyses confirmed the formation of inclusion complex. AE/HP-ß-CD showed the same antifungal activity of pure EA with MIC value of 25 µg/ml for C. albicans ATCC 18804 and 50 µg/ml for C. albicans SC 5314. Treatment with sub-MIC on C. albicans revealed increased MIC value in the presence of sorbitol for ATCC 18804 (50 µg/ml), suggesting activity on cell wall. Results suggest no effect on cell membrane. No effects of EA on morphogenesis and production of extracellular enzymes were detected. Protective effect of EA at 3.2, 6.4 e 32 µg/ml was observed in D. melanogaster model, resulting in survivals of 45, 33 and 34%. Low cytotoxicity was observed for concentrations of 200 and 250 µg/ml. In vivo tests in murine models indicated reduction in epithelial invasion after treatment EA/HP-ß-CD for 24 and 48 h when compared to control. In conclusion, EA/HP-ß-CD showed inhibitory effect on C. albicans in vitro, with low toxicity. Protective effect against fungal infection was observed in D. melanogaster model. In murine model of oral candidosis, EA/HP-ß-CD reduced fungal epithelial invasion(AU)


Asunto(s)
Humanos , Candida albicans/inmunología , Ciclodextrinas/efectos adversos , Ácido Elágico/administración & dosificación
18.
Rev chil anest ; 48(5): 461-464, 2019. tab
Artículo en Inglés | LILACS | ID: biblio-1509985

RESUMEN

OBJECTIVES: The capacity of polymorphonuclear cells (PMN) to phagocyte microorganisms is an important function to be preserved during surgical interventions. Therefore, the aim of this study was to determine the effect of propofol, fentanyl and remifentanil combination on Candida albicans engulfment by human PMN. MATERIALS AND METHODS: Twenty patients scheduled to undergo surgical interventions (ASA I-II) received propofol, fentanyl and remifentanil as intravenous anesthesia. PMNs were obtained before and after the surgical procedure and phagocytosis assay was performed using opsonized C. albicans. RESULTS AND CONCLUSIONS: No differences between the values obtained before and after anesthesia treatment in the number of phagocytic PMN and the number of C. albicans engulfed were observed. These results suggest that the used anesthesic protocol does not alter one of the most important immune mechanisms.


OBJETIVOS: La capacidad de las células polimorfonucleares (CPN) de fagocitar a los microorganismos es una importante función que se debe de preservar durante las intervenciones quirúrgicas. Por lo tanto, el propósito de este estudio fue determinar el efecto de la combinación del propofol, el fentanil y el remifentanil en la ingestión de Candida albicans por parte de las CPN humanas. MATERIALES Y MÉTODOS: Veinte pacientes sujetos a intervenciones quirúrgicas (ASA I-II) recibieron propofol, fentanil y remifentanil como anestesia endovenosa. Los CPN se obtuvieron antes y después del procedimiento quirúrgico y el ensayo de fagocitosis fue realizando usando C. albicans opsonizada. RESULTADOS Y CONCLUSIONES: No se observaron diferencias significativas en los valores obtenidos antes y después del tratamiento anestésico tanto en el número de CPN fagocíticas como en el número de C. albicans dentro de las células. Estos hallazgos sugieren que el protocolo anestésico usado no altera uno de los mecanismos de defensa más importante del organismo.


Asunto(s)
Humanos , Masculino , Femenino , Adulto , Persona de Mediana Edad , Candida albicans/inmunología , Propofol/administración & dosificación , Fentanilo/administración & dosificación , Anestésicos Intravenosos/administración & dosificación , Fagocitosis/efectos de los fármacos , Procedimientos Quirúrgicos Operativos , Propofol/farmacología , Fentanilo/farmacología , Anestésicos Intravenosos/farmacología , Quimioterapia Combinada , Remifentanilo , Neutrófilos
19.
São José dos Campos; s.n; 2019. 81 p. il., graf., tab..
Tesis en Portugués | BBO - Odontología, LILACS | ID: biblio-1146884

RESUMEN

Os probióticos são considerados uma alternativa potencial para o controle da candidose, no entanto, existe uma falta de produtos probióticos direcionados para a cavidade bucal. Neste estudo, desenvolvemos formulações probióticas usando gellan gum, um biopolímero natural usado como aditivo alimentar, e investigamos os efeitos dessas formulações em Candida albicans. Para isso, Lactobacillus paracasei 28.4, uma cepa recentemente isolada da cavidade bucal, foi incorporada em várias concentrações de gellan gum (1 a 0,6%). Todas as concentrações testadas foram capazes de incorporar as células de L. paracasei, mantendo a viabilidade bacteriana. As formulações probióticas permaneceram estáveis por 7 dias quando armazenadas à temperatura ambiente ou a 4°C. Entretanto, o armazenamento a longo prazo das formulações probióticas foi conseguido apenas quando L. paracasei 28.4 foi liofilizado. As formulações probióticas proporcionaram uma liberação de células de L. paracasei por 24 horas, o que foi suficiente para inibir o crescimento de C. albicans com efeitos dependentes das concentrações celulares incorporadas no gellan gum. As formulações probióticas também tiveram atividade inibitória contra os biofilmes de Candida, reduzindo o número de células de Candida (p<0,0001), diminuindo a biomassa total (p=0,0003) e prejudicando a formação de hifas (p=0,0002) em relação ao grupo controle não tratado. Contudo, a formulação probiótica de gellan gum a 1% proporcionou uma colonização oral de L. paracasei em camundongos com aproximadamente 6 log de UFC/mL após 10 dias. Essa formulação inibiu o crescimento de C. albicans (p<0,0001), impediu o desenvolvimento de lesões de candidose (p=0,0013) e suprimiu a inflamação (p = 0,0006) quando comparada aos camundongos não tratados. Estes resultados indicam que o gellan gum pode ser um biomaterial promissor como sistema transportador de probióticos para prevenir a candidose oral(AU)


Probiotics are considered a potential alternative for the control of candidiasis, however there is a lack of probiotic products targeted for the oral cavity. In this study, we developed probiotic formulations using gellan gum, a natural biopolymer used as a food-additive, and investigated the effects of this delivery method on Candida albicans. Lactobacillus paracasei 28.4, a strain recently isolated from the oral cavity, was incorporated in several concentrations of gellan gum (1 to 0.6%). All tested concentrations could incorporate L. paracasei cells while maintaining bacterial viability. Probiotic-gellan formulations were stable for 7 days when stored at room temperature or 4°C. Long-term storage of bacterial impregnated gellan gum could be achieved when L. paracasei 28.4 was lyophilized. The probiotic-gellan formulations provided a release of L. paracasei cells over 24 hours that was sufficient to inhibit the growth of C. albicans with effects dependent on the cell concentrations incorporated into gellan gum. The probiotic-gellan formulations also had inhibitory activity against Candida biofilms by reducing the number of Candida cells (p < 0.0001), decreasing the total biomass (p = 0.0003) and impairing hyphae formation (p = 0.0002) in relation to the control group, not treated. However, only probiotic formulation of gellan gum 1% provided an oral colonization of L. paracasei in mice with approximately 6 log of CFU/mL after 10 days. This formulation inhibited the C. albicans growth (p < 0.0001), prevented the development of candidiasis lesions (p = 0.0013), and suppressed the inflammation (p = 0.0006) when compared to the mice not treated. These results indicate that gellan gum is a promising biomaterial as a carrier system of probiotics to prevent oral candidiasis(AU)


Asunto(s)
Lacticaseibacillus paracasei/clasificación , Candida albicans/inmunología , Hidrogeles/administración & dosificación
20.
Microb Pathog ; 124: 21-29, 2018 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-30118801

RESUMEN

In recent years, C. albicans and C. glabrata have been identified as the main cause of candidemia and invasive candidiasis in hospitalized and immunocompromised patients. In order to colonize the human host, these fungi express several virulence factors such as the response to oxidative stress and the formation of biofilms. In the expression of these virulence factors, the cell wall of C. albicans and C. glabrata is of fundamental importance. As the outermost structure of the yeast, the cell wall is the first to come in contact with the reactive oxygen species (ROS) generated during the respiratory outbreak, and in the formation of biofilms, it is the first to adhere to organs or medical devices implanted in the human host. In both processes, several cell wall proteins (CWP) are required, since they promote attachment to human cells or abiotic surfaces, as well as to detoxify ROS. In our working group we have identified moonlighting CWP in response to oxidative stress as well as in the formation of biofilms. Having identified moonlighting CWP in Candida species in response to two virulence factors indicates that these proteins may possibly be immunodominant. The aim of the present work was to evaluate whether proteins of this type such as fructose-bisphosphate aldolase (Fba1), phosphoglycerate kinase (Pgk) and pyruvate kinase (Pk), could confer protection in a mouse model against C. albicans and C. glabrata. For this, recombinant proteins His6-Fba1, His6-Pgk and His6-Pk were constructed and used to immunize several groups of mice. The immunized mice were infected with C. albicans or C. glabrata, and subsequently the liver, spleen and kidney were extracted and the number of CFU was determined. Our results showed that Pk confers immunity to mice against C. albicans, while Fba1 to C. glabrata. This data allows us to conclude that the moonlighting CWP, Fba1 and Pk confer in vivo protection in a specific way against each species of Candida. This makes them promising candidates for developing specific vaccines against these pathogens.


Asunto(s)
Candidiasis/prevención & control , Fructosa-Bifosfato Aldolasa/inmunología , Proteínas Fúngicas/inmunología , Vacunas Fúngicas/inmunología , Fosfoglicerato Quinasa/inmunología , Piruvato Quinasa/inmunología , Animales , Candida albicans/inmunología , Candida glabrata/inmunología , Candidiasis/inmunología , Recuento de Colonia Microbiana , Modelos Animales de Enfermedad , Fructosa-Bifosfato Aldolasa/administración & dosificación , Proteínas Fúngicas/administración & dosificación , Vacunas Fúngicas/administración & dosificación , Riñón/microbiología , Hígado/microbiología , Ratones , Fosfoglicerato Quinasa/administración & dosificación , Piruvato Quinasa/administración & dosificación , Bazo/microbiología , Vacunas Sintéticas/administración & dosificación , Vacunas Sintéticas/inmunología
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