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1.
J Biol Chem ; 298(10): 102419, 2022 10.
Artigo em Inglês | MEDLINE | ID: mdl-36037968

RESUMO

Candida albicans (C. albicans) is a dimorphic commensal human fungal pathogen that can cause severe oropharyngeal candidiasis (oral thrush) in susceptible hosts. During invasive infection, C. albicans hyphae invade oral epithelial cells (OECs) and secrete candidalysin, a pore-forming cytolytic peptide that is required for C. albicans pathogenesis at mucosal surfaces. Candidalysin is produced in the hyphal invasion pocket and triggers cell damage responses in OECs. Candidalysin also activates multiple MAPK-based signaling events that collectively drive the production of downstream inflammatory mediators that coordinate downstream innate and adaptive immune responses. The activities of candidalysin are dependent on signaling through the epidermal growth factor receptor (EGFR). Here, we interrogated known EGFR-MAPK signaling intermediates for their roles mediating the OEC response to C. albicans infection. Using RNA silencing and pharmacological inhibition, we identified five key adaptors, including growth factor receptor-bound protein 2 (Grb2), Grb2-associated binding protein 1 (Gab1), Src homology and collagen (Shc), SH2-containing protein tyrosine phosphatase-2 (Shp2), and casitas B-lineage lymphoma (c-Cbl). We determined that all of these signaling effectors were inducibly phosphorylated in response to C. albicans. These phosphorylation events occurred in a candidalysin-dependent manner and additionally required EGFR phosphorylation, matrix metalloproteinases (MMPs), and cellular calcium flux to activate a complete OEC response to fungal infection. Of these, Gab1, Grb2, and Shp2 were the dominant drivers of ERK1/2 activation and the subsequent production of downstream innate-acting cytokines. Together, these results identify the key adaptor proteins that drive the EGFR signaling mechanisms that underlie oral epithelial responses to C. albicans.


Assuntos
Candida albicans , Candidíase Bucal , Receptores ErbB , Proteínas Fúngicas , Mucosa Bucal , Humanos , Candida albicans/metabolismo , Candida albicans/patogenicidade , Citocinas/metabolismo , Receptores ErbB/metabolismo , Proteínas Fúngicas/metabolismo , Proteínas Adaptadoras da Sinalização Shc/metabolismo , Candidíase Bucal/metabolismo , Candidíase Bucal/microbiologia , Mucosa Bucal/metabolismo , Mucosa Bucal/microbiologia , Células Epiteliais/metabolismo , Células Epiteliais/microbiologia
2.
Int J Mol Sci ; 23(4)2022 Feb 13.
Artigo em Inglês | MEDLINE | ID: mdl-35216181

RESUMO

Resistance to antifungal therapy of Candida albicans and non-albicans Candida strains, frequently associated with oral candidosis, is on the rise. In this context, host-defense peptides have emerged as new promising candidates to overcome antifungal resistance. Thus, the aim of this study was to assess the effectiveness against Candida species of different Catestatin-derived peptides, as well as the combined effect with serum albumin. Among Catestatin-derived peptides, the most active against sensitive and resistant strains of C. albicans, C. tropicalis and C. glabrata was the D-isomer of Cateslytin (D-bCtl) whereas the efficiency of the L-isomer (L-bCtl) significantly decreases against C. glabrata strains. Images obtained by transmission electron microscopy clearly demonstrated fungal membrane lysis and the leakage of the intracellular material induced by the L-bCtl and D-bCtl peptides. The possible synergistic effect of albumin on Catestatin-derived peptides activity was investigated too. Our finding showed that bovine serum albumin (BSA) when combined with the L- isomer of Catestatin (L-bCts) had a synergistic effect against Candida albicans especially at low concentrations of BSA; however, no synergistic effect was detected when BSA interacted with L-bCtl, suggesting the importance of the C-terminal end of L-bCts (GPGLQL) for the interaction with BSA. In this context in vitro D-bCtl, as well as the combination of BSA with L-bCts are potential candidates for the development of new antifungal drugs for the treatment of oral candidosis due to Candida and non-Candida albicans, without detrimental side effects.


Assuntos
Candidíase Bucal/tratamento farmacológico , Cromogranina A/farmacologia , Fragmentos de Peptídeos/farmacologia , Peptídeos/farmacologia , Animais , Antifúngicos/farmacologia , Candida/efeitos dos fármacos , Candida/metabolismo , Candidíase Bucal/metabolismo , Bovinos , Farmacorresistência Fúngica/efeitos dos fármacos , Humanos , Soroalbumina Bovina/metabolismo
3.
J Clin Lab Anal ; 36(2): e24208, 2022 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-34997991

RESUMO

INTRODUCTION: Resistance to azole drugs has been observed in candidiasis due to their long-term use and poor response to treatment. Resistance to azole drugs in Candida albicans isolates is controlled by several genes including ERG11, CDR1, CDR2, and MDR1. In this study, the expression of the mentioned genes was evaluated in C. albicans isolates susceptible and resistant to fluconazole. METHODS: After identifying the Candida isolates using morphological and molecular methods, the minimum inhibitory concentration (MIC) and drug susceptibility were determined using the European Committee on Antimicrobial Susceptibility Testing (EUCAST) method. RNA was then extracted and cDNA was synthesized from 24 C. albicans isolates from patients with cancer. Then, the mean expressions of these genes were compared in two groups using real-time polymerase chain reaction (RT-PCR). RESULTS: A total of 74 Candida isolates were obtained from the oral cavity of 61 cancer patients with oral candidiasis. After 24 h, 21.6% of the isolates were fluconazole-resistant, 10.8% were identified as dose-dependent, and the rest of the isolates (67.6%) were fluconazole-sensitive. The mean expressions of the CDR1 and MDR1 genes were significantly higher in the resistant isolates than in the sensitive ones. However, the ERG11 and CDR2 genes were not significantly increased in the resistant isolates. CONCLUSION: The increased mean expressions of the CDR1 and MDR1 genes had a greater effect on fluconazole resistance among the drug-resistant strains of C. albicans in chemotherapy patients. It seemed that the accumulation of chemotherapeutic drugs in this organism stimulated some regulatory factors and increased the expression of these two genes and ultimately helped to further increase their expression and resistance to fluconazole.


Assuntos
Candida albicans/genética , Candidíase Bucal/metabolismo , Farmacorresistência Fúngica/genética , Fluconazol/farmacologia , Proteínas Fúngicas/genética , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/genética , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismo , Antifúngicos/farmacologia , Candida albicans/efeitos dos fármacos , Candida albicans/isolamento & purificação , Candidíase Bucal/etiologia , Proteínas Fúngicas/metabolismo , Expressão Gênica , Humanos , Irã (Geográfico) , Proteínas de Membrana Transportadoras/genética , Proteínas de Membrana Transportadoras/metabolismo , Neoplasias/complicações , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Esterol 14-Desmetilase/genética , Esterol 14-Desmetilase/metabolismo
4.
mBio ; 12(6): e0271621, 2021 12 21.
Artigo em Inglês | MEDLINE | ID: mdl-34724825

RESUMO

During oropharyngeal candidiasis, Candida albicans activates the epidermal growth factor receptor (EGFR), which induces oral epithelial cells to endocytose the fungus and synthesize proinflammatory mediators. To elucidate EGFR signaling pathways that are stimulated by C. albicans, we used proteomics to identify 1,214 proteins that were associated with EGFR in C. albicans-infected cells. Seven of these proteins were selected for additional study. Among these proteins, WW domain-binding protein 2, Toll-interacting protein, interferon-induced transmembrane protein 3 (IFITM3), and the globular C1q receptor (gC1qR) were found to associate with EGFR in viable oral epithelial cells. Each of these proteins was required for maximal endocytosis of C. albicans, and all regulated fungus-induced production of interleukin-1ß (IL-1ß) and/or IL-8, either positively or negatively. gC1qR was found to function as a key coreceptor with EGFR. Interacting with the C. albicans Als3 invasin, gC1qR was required for the fungus to induce autophosphorylation of both EGFR and the ephrin type A receptor 2. The combination of gC1qR and EGFR was necessary for maximal endocytosis of C. albicans and secretion of IL-1ß, IL-8, and granulocyte-macrophage colony-stimulating factor (GM-CSF) by human oral epithelial cells. In mouse oral epithelial cells, inhibition of gC1qR failed to block C. albicans-induced phosphorylation, and knockdown of IFITM3 did not inhibit C. albicans endocytosis, indicating that gC1qR and IFITM3 function differently in mouse versus human oral epithelial cells. Thus, this work provides an atlas of proteins that associate with EGFR and identifies several that play a central role in the response of human oral epithelial cells to C. albicans infection. IMPORTANCE Oral epithelial cells play a key role in the pathogenesis of oropharyngeal candidiasis. In addition to being target host cells for C. albicans adherence and invasion, they secrete proinflammatory cytokines and chemokines that recruit T cells and activated phagocytes to foci of infection. It is known that C. albicans activates EGFR on oral epithelial cells, which induces these cells to endocytose the organism and stimulates them to secrete proinflammatory mediators. To elucidate the EGFR signaling pathways that govern these responses, we analyzed the epithelial cell proteins that associate with EGFR in C. albicans-infected epithelial cells. We identified four proteins that physically associate with EGFR and that regulate different aspects of the epithelial response to C. albicans. One of these is gC1qR, which is required for C. albicans to activate EGFR, induce endocytosis, and stimulate the secretion of proinflammatory mediators, indicating that gC1qR functions as a key coreceptor with EGFR.


Assuntos
Candida albicans/fisiologia , Candidíase Bucal/metabolismo , Receptores ErbB/metabolismo , Glicoproteínas de Membrana/metabolismo , Receptores de Complemento/metabolismo , Animais , Candidíase Bucal/genética , Candidíase Bucal/microbiologia , Células Epiteliais/metabolismo , Células Epiteliais/microbiologia , Receptores ErbB/genética , Humanos , Glicoproteínas de Membrana/genética , Camundongos , Camundongos Endogâmicos BALB C , Células NIH 3T3 , Ligação Proteica , Receptores de Complemento/genética , Transdução de Sinais
5.
J Cell Mol Med ; 25(19): 9473-9475, 2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-34486221

RESUMO

While cigarette smoke compounds are known to have immunosuppressive effects on the oral mucosa, the relationship between in vivo immune dysfunction caused by smoking and the development of oral Candida infections remains largely unexplored. In a recent issue of The Journal of Cellular and Molecular Medicine, Ye and colleagues provide evidence that smoking increases oral mucosa susceptibility to Candida albicans infection via the activation of the Nrf2 pathway, which in turn negatively regulates the NLRP3 inflammasome. This opens new perspective in considering Nrf2 as a relevant target for smoking-induced C. albicans-related oral diseases.


Assuntos
Candidíase Bucal/etiologia , Candidíase Bucal/metabolismo , Inflamassomos/metabolismo , Fator 2 Relacionado a NF-E2/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Fumar/efeitos adversos , Biomarcadores , Candida albicans , Suscetibilidade a Doenças , Humanos , Modelos Biológicos , Mucosa Bucal/metabolismo , Mucosa Bucal/microbiologia , Estresse Oxidativo , Espécies Reativas de Oxigênio/metabolismo
6.
PLoS Pathog ; 17(1): e1009221, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-33471869

RESUMO

During oropharyngeal candidiasis (OPC), Candida albicans invades and damages oral epithelial cells, which respond by producing proinflammatory mediators that recruit phagocytes to foci of infection. The ephrin type-A receptor 2 (EphA2) detects ß-glucan and plays a central role in stimulating epithelial cells to release proinflammatory mediators during OPC. The epidermal growth factor receptor (EGFR) also interacts with C. albicans and is known to be activated by the Als3 adhesin/invasin and the candidalysin pore-forming toxin. Here, we investigated the interactions among EphA2, EGFR, Als3 and candidalysin during OPC. We found that EGFR and EphA2 constitutively associate with each other as part of a heteromeric physical complex and are mutually dependent for C. albicans-induced activation. Als3-mediated endocytosis of a C. albicans hypha leads to the formation of an endocytic vacuole where candidalysin accumulates at high concentration. Thus, Als3 potentiates targeting of candidalysin, and both Als3 and candidalysin are required for C. albicans to cause maximal damage to oral epithelial cells, sustain activation of EphA2 and EGFR, and stimulate pro-inflammatory cytokine and chemokine secretion. In the mouse model of OPC, C. albicans-induced production of CXCL1/KC and CCL20 is dependent on the presence of candidalysin and EGFR, but independent of Als3. The production of IL-1α and IL-17A also requires candidalysin but is independent of Als3 and EGFR. The production of TNFα requires Als1, Als3, and candidalysin. Collectively, these results delineate the complex interplay among host cell receptors EphA2 and EGFR and C. albicans virulence factors Als1, Als3 and candidalysin during the induction of OPC and the resulting oral inflammatory response.


Assuntos
Candida albicans/fisiologia , Candidíase Bucal/patologia , Efrina-A2/metabolismo , Células Epiteliais/patologia , Orofaringe/patologia , Fatores de Virulência/metabolismo , Animais , Candidíase Bucal/genética , Candidíase Bucal/metabolismo , Candidíase Bucal/microbiologia , Citocinas/metabolismo , Modelos Animais de Doenças , Efrina-A2/genética , Células Epiteliais/metabolismo , Células Epiteliais/microbiologia , Receptores ErbB/genética , Receptores ErbB/metabolismo , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Orofaringe/metabolismo , Orofaringe/microbiologia , Receptor EphA2 , Fatores de Virulência/genética
7.
Methods Mol Biol ; 2260: 133-143, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33405035

RESUMO

Microbial interactions with epithelial barriers are important steps preceding disease. Infections with Candida albicans are no exception. This opportunistic fungus, commonly harmlessly residing in close proximity to human epithelia, can shift to a more pathogenic form, can invade tissues, and cause disease. Pathogenesis, in C. albicans as well as in many other microorganisms, is characterized by three important steps: adhesion to-, invasion into-, and damage of host cells. In this book chapter, we describe three well-established protocols that allow us to differentially stain C. albicans cells adhering to and invading into host cells, therefore allowing quantifications of such processes. We also describe a common host cell cytotoxicity assay that employs a commercial kit, adapted to C. albicans.


Assuntos
Candida albicans/patogenicidade , Candidíase Bucal/microbiologia , Adesão Celular , Células Epiteliais/microbiologia , Microscopia de Fluorescência , Mucosa Bucal/microbiologia , Candidíase Bucal/metabolismo , Candidíase Bucal/patologia , Linhagem Celular , Sobrevivência Celular , Células Epiteliais/metabolismo , Células Epiteliais/patologia , Interações Hospedeiro-Patógeno , Humanos , Mucosa Bucal/metabolismo , Mucosa Bucal/patologia
8.
Nat Microbiol ; 3(1): 53-61, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-29133884

RESUMO

Oral epithelial cells discriminate between pathogenic and non-pathogenic stimuli, and only induce an inflammatory response when they are exposed to high levels of a potentially harmful microorganism. The pattern recognition receptors (PRRs) in epithelial cells that mediate this differential response are poorly understood. Here, we demonstrate that the ephrin type-A receptor 2 (EphA2) is an oral epithelial cell PRR that binds to exposed ß-glucans on the surface of the fungal pathogen Candida albicans. Binding of C. albicans to EphA2 on oral epithelial cells activates signal transducer and activator of transcription 3 and mitogen-activated protein kinase signalling in an inoculum-dependent manner, and is required for induction of a proinflammatory and antifungal response. EphA2 -/- mice have impaired inflammatory responses and reduced interleukin-17 signalling during oropharyngeal candidiasis, resulting in more severe disease. Our study reveals that EphA2 functions as a PRR for ß-glucans that senses epithelial cell fungal burden and is required for the maximal mucosal inflammatory response to C. albicans.


Assuntos
Candida albicans/metabolismo , Candidíase Bucal/metabolismo , Mucosa Bucal/metabolismo , Receptor EphA2/metabolismo , Receptores de Reconhecimento de Padrão/metabolismo , beta-Glucanas/metabolismo , Animais , Candida albicans/crescimento & desenvolvimento , Candidíase Bucal/patologia , Linhagem Celular , Citocinas/biossíntese , Modelos Animais de Doenças , Endocitose , Células Epiteliais/citologia , Células Epiteliais/metabolismo , Células Epiteliais/microbiologia , Receptores ErbB/antagonistas & inibidores , Receptores ErbB/metabolismo , Mediadores da Inflamação/análise , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Mucosa Bucal/citologia , Mucosa Bucal/microbiologia , Fosforilação , Receptor EphA2/antagonistas & inibidores , Receptor EphA2/deficiência , Receptores de Reconhecimento de Padrão/antagonistas & inibidores , Receptores de Reconhecimento de Padrão/deficiência , Fator de Transcrição STAT3/metabolismo , Transdução de Sinais
9.
Oral Dis ; 23(7): 941-948, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-28403570

RESUMO

OBJECTIVE: To assess changes in the salivary expression of IL-1α, IL-1ß, IL-2, IL-6, IL-10, IL-17, and TNF in acute leukemia (AL) patients before and during chemotherapy, and its association with HSV infection, oral candidiasis (OC), and oral mucositis (OM) onset. METHODS: Cohort study in AL patients >15 years starting induction chemotherapy at a Mexican oncological center (2013-2014). Onset of oral lesions (OLs) was assessed during follow-up, and saliva was obtained at baseline, at visit 2 (days 4-12), and at visit 3 (days 13-21) after chemotherapy, treated with a protease inhibitor and stored at -70°C. An enzyme-linked immunosorbent assay was performed. Cox proportional hazards regression models were constructed to estimate hazard ratios and its 95% CI (HR, 95% CI) for OL development. RESULTS: Forty-one patients were followed up, and 17 (41.5%) developed OLs. OL patients had higher baseline salivary IL-1α than those without lesions (p = 0.040). During visit 2, OL patients had higher levels of IL-1α (p = 0.033), IL-1ß (p = 0.016), IL-6 (p = 0.035), and TNF (p = 0.019) than those who did not develop OLs. Patients with HSV infection, OC, and OM showed higher salivary TNF levels during follow-up (HR: 3.52, 95% CI: 1.35-9.14, p = 0.010). CONCLUSION: AL patients undergoing chemotherapy with high salivary TNF levels were more likely to develop HSV infection, OC, and OM.


Assuntos
Candidíase Bucal/metabolismo , Citocinas/metabolismo , Herpes Simples/metabolismo , Saliva/metabolismo , Estomatite/metabolismo , Adulto , Antineoplásicos/efeitos adversos , Biomarcadores/metabolismo , Candidíase Bucal/diagnóstico , Doxiciclina/efeitos adversos , Feminino , Herpes Simples/diagnóstico , Humanos , Leucemia/tratamento farmacológico , Masculino , Ensaios Clínicos Controlados Aleatórios como Assunto , Estomatite/diagnóstico , Estomatite/etiologia , Fator de Necrose Tumoral alfa/metabolismo , Adulto Jovem
10.
Oral Dis ; 22(8): 805-814, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27495361

RESUMO

OBJECTIVES: To compare biofilm-forming ability, hydrolytic enzymes and ethanol-derived acetaldehyde production of oral Candida isolated from the patients with oral cancer and matched non-oral cancer. MATERIAL AND METHODS: Fungal biofilms were grown in RPMI-1640 medium, and biofilm mass and biofilm activity were assessed using crystal violet staining and XTT salt reduction assays, respectively. Phospholipase, proteinase, and esterase production were measured using agar plate method, while fungal acetaldehyde production was assessed via gas chromatography. RESULTS: Candida isolated from patients with oral cancer demonstrated significantly higher biofilm mass (P = 0.031), biofilm metabolic activity (P < 0.001), phospholipase (P = 0.002), and proteinase (P = 0.0159) activity than isolates from patients with non-oral cancer. High ethanol-derived acetaldehyde-producing Candida were more prevalent in patients with oral cancer than non-oral cancer (P = 0.01). In univariate regression analysis, high biofilm mass (P = 0.03) and biofilm metabolic activity (P < 0.001), high phospholipase (P = 0.003), and acetaldehyde production ability (0.01) were significant risk factors for oral cancer; while in the multivariate regression analysis, high biofilm activity (0.01) and phospholipase (P = 0.01) were significantly positive influencing factors on oral cancer. CONCLUSION: These data suggest a significant positive association between the ability of Candida isolates to form biofilms, to produce hydrolytic enzymes, and to metabolize alcohol to acetaldehyde with their ability to promote oral cancer development.


Assuntos
Acetaldeído/metabolismo , Candida/patogenicidade , Candidíase Bucal/microbiologia , Neoplasias Bucais/microbiologia , Biofilmes/crescimento & desenvolvimento , Candida/metabolismo , Candidíase Bucal/metabolismo , Estudos de Casos e Controles , Etanol/metabolismo , Feminino , Humanos , Masculino
11.
Oral Dis ; 22(1): 69-74, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26577981

RESUMO

OBJECTIVES: Candida albicans attaches to oral surfaces via a number of mechanisms including adherence mediated by salivary components adsorbed to the C. albicans cell surface. Our goal was to identify the salivary molecules involved. MATERIALS AND METHODS: Biotinylated salivary polypeptides that were bound by C. albicans were detected in extracts from washed, saliva-treated yeast cells by polyacrylamide gel electrophoresis and electroblot or immunoblot transfer analysis and purified by electroelution. Purified material was tested for the ability to promote the adherence of radiolabelled C. albicans yeast cells to cultured epithelial monolayers. RESULTS: Three of the polypeptides bound by C. albicans cells were identified as components of secretory IgA, including secretory component. Using non-denaturing polyacrylamide gel electrophoresis, we demonstrated that secretory component could be detected in its free form in saliva, and was bound by yeast cells. Secretory component which was purified by electroelution from non-denaturing PAGE-separated saliva, without detectable complete IgA, promoted adherence of yeast cells to cultured epithelial monolayers in a dose-dependent fashion. CONCLUSION: These results indicate that despite the inhibitory effect on adherence of IgA specific to C. albicans, IgA components, in particular secretory component, also promote binding to cultured epithelial monolayers.


Assuntos
Candida albicans/metabolismo , Células Epiteliais/microbiologia , Componente Secretório/metabolismo , Biotinilação , Candidíase Bucal/metabolismo , Candidíase Bucal/microbiologia , Adesão Celular/fisiologia , Linhagem Celular , Eletroforese em Gel de Poliacrilamida , Humanos , Imunoglobulina A Secretora/química , Imunoglobulina A Secretora/metabolismo , Mucosa Bucal/química , Mucosa Bucal/metabolismo , Mucosa Bucal/microbiologia , Peptídeos/química , Complexo Glicoproteico GPIb-IX de Plaquetas/química , Complexo Glicoproteico GPIb-IX de Plaquetas/metabolismo , Saliva/química , Saliva/metabolismo
12.
Med Pregl ; 67(5-6): 149-53, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25033573

RESUMO

INTRODUCTION: Candidiasis has become a human disease of increasing importance in the last decades. The aim of the study is to establish pathomorphological alterations caused by the blastospores of the Candida albicans as well as morphometric alterations. MATERIAL AND METHODS: The experiment was carried out on 2.5-month-old rats, weighting 110-130 g. The study sample was divided into the animals infected by a submucous inoculation in the periodontal region and the controls. The gingival specimens were taken, preparations were done and stained by the hematoxylin-eosin and Periodic acid Schiff methods. RESULTS: The following alterations were found out by the stereological analysis: an average volume of nuclei of the gingival epithelial cells was 111.82 microm3 (SD = 25.34) on the first day. A statistically significant increase in the volume of nuclei in the experimental group began to occur from the fourth day (202.97 microm3; SD = 31.16, p < 0.05) and the highest value of the nuclei volume was found out on the eight day of the experiment (316.83 microm3; SD = 40.15). CONCLUSION: Blastospores of Candida albicans are pathogenic for the gingival tissue where they cause degenerative necrotic alterations of the granulomatous character and after the fourth day from the inoculation, the development of the pseudohyphae was observed. The obtained values of stereologic measurement show the acute increase in the volume of nuclei.


Assuntos
Candida albicans/crescimento & desenvolvimento , Candidíase Bucal/microbiologia , Modelos Animais de Doenças , Células Epiteliais/microbiologia , Gengiva/microbiologia , Animais , Candida albicans/metabolismo , Candidíase Bucal/metabolismo , Contagem de Colônia Microbiana , Células Epiteliais/metabolismo , Gengiva/metabolismo , Ratos
13.
Infect Immun ; 82(3): 1030-5, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24343647

RESUMO

Oropharyngeal candidiasis (OPC; thrush) is an opportunistic fungal infection caused by the commensal microbe Candida albicans. Immunity to OPC is strongly dependent on CD4+ T cells, particularly those of the Th17 subset. Interleukin-17 (IL-17) deficiency in mice or humans leads to chronic mucocutaneous candidiasis, but the specific downstream mechanisms of IL-17-mediated host defense remain unclear. Lipocalin 2 (Lcn2; 24p3; neutrophil gelatinase-associated lipocalin [NGAL]) is an antimicrobial host defense factor produced in response to inflammatory cytokines, particularly IL-17. Lcn2 plays a key role in preventing iron acquisition by bacteria that use catecholate-type siderophores, and lipocalin 2(-/-) mice are highly susceptible to infection by Escherichia coli and Klebsiella pneumoniae. The role of Lcn2 in mediating immunity to fungi is poorly defined. Accordingly, in this study, we evaluated the role of Lcn2 in immunity to oral infection with C. albicans. Lcn2 is strongly upregulated following oral infection with C. albicans, and its expression is almost entirely abrogated in mice with defective IL-17 signaling (IL-17RA(-/-) or Act1(-/-) mice). However, Lcn2(-/-) mice were completely resistant to OPC, comparably to wild-type (WT) mice. Moreover, Lcn2 deficiency mediated protection from OPC induced by steroid immunosuppression. Therefore, despite its potent regulation during C. albicans infection, Lcn2 is not required for immunity to mucosal candidiasis.


Assuntos
Proteínas de Fase Aguda/metabolismo , Candidíase Bucal/imunologia , Candidíase Bucal/metabolismo , Interleucina-17/metabolismo , Lipocalinas/metabolismo , Proteínas Oncogênicas/metabolismo , Proteínas de Fase Aguda/genética , Proteínas de Fase Aguda/imunologia , Animais , Candida albicans/imunologia , Candidíase Bucal/genética , Candidíase Bucal/microbiologia , Interleucina-17/genética , Interleucina-17/imunologia , Lipocalina-2 , Lipocalinas/genética , Lipocalinas/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Mucosa Bucal/imunologia , Mucosa Bucal/metabolismo , Mucosa Bucal/microbiologia , Mucosa/imunologia , Mucosa/metabolismo , Mucosa/microbiologia , Proteínas Oncogênicas/genética , Proteínas Oncogênicas/imunologia , Receptores de Interleucina-17/genética , Receptores de Interleucina-17/imunologia , Receptores de Interleucina-17/metabolismo , Regulação para Cima/genética , Regulação para Cima/imunologia
14.
Mol Biosyst ; 8(12): 3216-23, 2012 Oct 30.
Artigo em Inglês | MEDLINE | ID: mdl-23041753

RESUMO

Denture stomatitis (DS) is the most common oral pathology among denture wearers, affecting over one-third of this group. DS is usually associated with C. albicans. However, unlike other oral candidiasis, most DS patients have intact host immunity. The presence of a denture alone is usually sufficient for DS. Saliva and its protein contents can theoretically predispose some denture wearers to DS and others resistant toward DS. Here we proposed for the first time to define salivary proteomic profiles of denture wearers with and without DS. SELDI-TOF/MS analysis suggests that there is a proteomic differentiation among control, localized and generalized DS. Based on initial SELDI-TOF/MS profiling, we further used reversed phase liquid chromatography, MALDI-TOF/MS, and LC-MS/MS to characterize the salivary proteins associated with DS. Nineteen proteins based on SELDI-TOF/MS profiling were found including cystatin-SN, statherin, kininogen-1, desmocollin-2, carbonic anhydrase-6, peptidyl-prolyl cis-trans isomerase A like peptides, cystatin C, and several immunoglobulin fragments. The proteomic content gives evidence of the interaction between host tissue, saliva, and candida. Further examination in larger populations of these proteins may help to gain a better understanding of DS pathological processes and improve DS treatments.


Assuntos
Dentaduras/efeitos adversos , Proteínas e Peptídeos Salivares/metabolismo , Estomatite sob Prótese/etiologia , Estomatite sob Prótese/metabolismo , Idoso , Candida albicans/imunologia , Candida albicans/metabolismo , Candida albicans/patogenicidade , Candidíase Bucal/imunologia , Candidíase Bucal/metabolismo , Feminino , Humanos , Imunoglobulinas/análise , Masculino , Pessoa de Meia-Idade , Análise Serial de Proteínas , Proteômica , Saliva/química , Saliva/imunologia , Saliva/metabolismo , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Estomatite sob Prótese/imunologia , Estomatite sob Prótese/microbiologia
15.
Proc Natl Acad Sci U S A ; 109(35): 14194-9, 2012 Aug 28.
Artigo em Inglês | MEDLINE | ID: mdl-22891338

RESUMO

The fungus Candida albicans is the major cause of oropharyngeal candidiasis (OPC). A key feature of this disease is fungal invasion of oral epithelial cells, a process that can occur by active penetration and fungal-induced endocytosis. Two invasins, Als3 and Ssa1, induce epithelial cell endocytosis of C. albicans, in part by binding to E-cadherin. However, inhibition of E-cadherin function only partially reduces C. albicans endocytosis, suggesting that there are additional epithelial cell receptors for this organism. Here, we show that the EGF receptor (EGFR) and HER2 function cooperatively to induce the endocytosis of C. albicans hyphae. EGFR and HER2 interact with C. albicans in an Als3- and Ssa1-dependent manner, and this interaction induces receptor autophosphorylation. Signaling through both EGFR and HER2 is required for maximal epithelial cell endocytosis of C. albicans in vitro. Importantly, oral infection with C. albicans stimulates the phosphorylation of EGFR and HER2 in the oral mucosa of mice, and treatment with a dual EGFR and HER2 kinase inhibitor significantly decreases this phosphorylation and reduces the severity of OPC. These results show the importance of EGFR and HER2 signaling in the pathogenesis of OPC and indicate the feasibility of treating candidal infections by targeting the host cell receptors with which the fungus interacts.


Assuntos
Candida albicans/metabolismo , Candidíase Bucal/metabolismo , Receptores ErbB/metabolismo , Mucosa Bucal/microbiologia , Receptor ErbB-2/metabolismo , Transdução de Sinais/fisiologia , Adenosina Trifosfatases/metabolismo , Animais , Caderinas/metabolismo , Candida albicans/crescimento & desenvolvimento , Candidíase Bucal/patologia , Modelos Animais de Doenças , Endocitose/fisiologia , Células Epiteliais/citologia , Células Epiteliais/metabolismo , Células Epiteliais/microbiologia , Proteínas Fúngicas/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Mucosa Bucal/citologia , Mucosa Bucal/metabolismo , Células NIH 3T3 , Fosforilação/fisiologia , Tirosina/metabolismo
16.
J Oral Pathol Med ; 41(10): 769-78, 2012 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-22553989

RESUMO

BACKGROUND: Oral epithelial cells significantly influence host inflammatory responses against Candida albicans in oropharyngeal candidiasis. Pro-inflammatory cytokines function as an early innate immune system mediator during C. albicans infection in oral epithelial cells. We sought to elucidate the pattern of the molecular mechanisms governing the human gingival epithelial cells (HGECs) to C. albicans infection likely involve multiple converging signal transduction pathways. MATERIALS AND METHODS: Primary HGECs were cultured with C. albicans ATCC90029. Total RNA was extracted after 8 h of infection and monitored mRNA levels using Affymetrix GeneChip (Human Genome U133 plus 2.0 Array, 48 000 genes). GeneChip data was analyzed by GeneSpring software and Ingenuity Pathway Analysis system. Reverse transcription polymerase chain reaction (RT-PCR), real-time RT-PCR and immunohistochemistry were used to investigate gene expression changes. RESULTS: The differentially expressed genes represented functions as diverse as immune response and inflammatory disease. IL-8, ICAM-1 and Cox-2 showed a greater than two fold change in expression relative to those in control cells. Altered mRNA levels in GeneChip analysis were confirmed by RT-PCR and real-time RT-PCR. Stronger immunoreactivity against ICAM-1 and Cox-2 was also observed in the infection with C. albicans in rat gingival epithelium. We have identified differential gene expression up-regulated or down-regulated with the up-regulation of IL-8 in C. albicans-treated cells. CONCLUSION: These findings indicate that the molecular mechanisms underlying the IL-8 response of HGECs to C. albicans infection likely involve multiple converging signal transduction pathways.


Assuntos
Candidíase Bucal/metabolismo , Perfilação da Expressão Gênica , Gengiva/metabolismo , Interleucina-8/metabolismo , Transdução de Sinais/genética , Animais , Candidíase Bucal/imunologia , Ciclo-Oxigenase 2/metabolismo , Células Epiteliais/citologia , Células Epiteliais/metabolismo , Gengiva/citologia , Humanos , Molécula 1 de Adesão Intercelular/metabolismo , Masculino , Análise de Sequência com Séries de Oligonucleotídeos , RNA/análise , RNA Mensageiro/análise , Ratos , Ratos Sprague-Dawley
17.
J Investig Clin Dent ; 3(3): 176-81, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22489001

RESUMO

AIM: To examine the presence of markers associated with malignancy, including p53, p21 cyclin-dependent kinase inhibitor 1A, murine double minutes-2, and others, in chronic hyperplastic candidiasis. METHODS: Immunohistochemical methods were used to examine the expression of p53, murine double minutes-2, p21 cyclin-dependent kinase inhibitor 1A, metallothionein, and proliferating cell nuclear antigen in 42 chronic hyperplastic candidiasis lesions and 11 non-infected control tissues. Terminal deoxynucleotidyl transferase-mediated digoxigenin-dUTP nick-end labeling was used to examine apoptosis, which was correlated with p53 expression. These markers were measured in lesions of chronic hyperplastic candidiasis that did not show any epithelial dysplasia or histological signs of malignancy. RESULTS: p53 scores were higher in chronic hyperplastic candidiasis than in controls (P = 0.0046). Murine double-minutes 2 levels were not elevated. p21 cyclin-dependent kinase inhibitor 1A was increased in parabasal (P < 0.0001) and basal epithelial cells. Chronic hyperplastic candidiasis lesions showed a similar basal/parabasal metallothionein staining pattern to that seen in normal squamous epithelium. Proliferating cell nuclear antigen was increased (P = 0.0007), as was apoptosis (P = 0.0033). CONCLUSION: Increased p53 in oral chronic hyperplastic candidiasis suggests an increased potential for malignant change in the epithelium, above that of normal tissues. Further functional investigation is required, as well as clinical follow-up studies.


Assuntos
Biomarcadores Tumorais/metabolismo , Candidíase Bucal/complicações , Carcinoma de Células Escamosas/etiologia , Hiperplasia/metabolismo , Mucosa Bucal/metabolismo , Neoplasias Bucais/metabolismo , Proteína Supressora de Tumor p53/metabolismo , Biomarcadores Tumorais/genética , Candidíase Bucal/metabolismo , Candidíase Bucal/patologia , Carcinoma de Células Escamosas/patologia , Inibidor de Quinase Dependente de Ciclina p21/genética , Inibidor de Quinase Dependente de Ciclina p21/metabolismo , Células Epiteliais/metabolismo , Células Epiteliais/patologia , Feminino , Expressão Gênica , Humanos , Hiperplasia/patologia , Imuno-Histoquímica , Marcação In Situ das Extremidades Cortadas , Masculino , Metalotioneína/genética , Metalotioneína/metabolismo , Pessoa de Meia-Idade , Mucosa Bucal/patologia , Neoplasias Bucais/patologia , Antígeno Nuclear de Célula em Proliferação/genética , Antígeno Nuclear de Célula em Proliferação/metabolismo , Proteínas Proto-Oncogênicas c-mdm2/genética , Proteínas Proto-Oncogênicas c-mdm2/metabolismo , Proteína Supressora de Tumor p53/genética
18.
BMC Complement Altern Med ; 12: 6, 2012 Jan 16.
Artigo em Inglês | MEDLINE | ID: mdl-22248145

RESUMO

BACKGROUND: Oral candidiasis is a common fungal disease mainly caused by Candida albicans. The aim of this study was to investigate the effects of A-type cranberry proanthocyanidins (AC-PACs) on pathogenic properties of C. albicans as well as on the inflammatory response of oral epithelial cells induced by this oral pathogen. METHODS: Microplate dilution assays were performed to determine the effect of AC-PACs on C. albicans growth as well as biofilm formation stained with crystal violet. Adhesion of FITC-labeled C. albicans to oral epithelial cells and to acrylic resin disks was monitored by fluorometry. The effects of AC-PACs on C. albicans-induced cytokine secretion, nuclear factor-kappa B (NF-κB) p65 activation and kinase phosphorylation in oral epithelial cells were determined by immunological assays. RESULTS: Although AC-PACs did not affect growth of C. albicans, it prevented biofilm formation and reduced adherence of C. albicans to oral epithelial cells and saliva-coated acrylic resin discs. In addition, AC-PACs significantly decreased the secretion of IL-8 and IL-6 by oral epithelial cells stimulated with C. albicans. This anti-inflammatory effect was associated with reduced activation of NF-κB p65 and phosphorylation of specific signal intracellular kinases. CONCLUSION: AC-PACs by affecting the adherence properties of C. albicans and attenuating the inflammatory response induced by this pathogen represent potential novel therapeutic agents for the prevention/treatment of oral candidiasis.


Assuntos
Candida albicans/efeitos dos fármacos , Candidíase Bucal/tratamento farmacológico , Interleucinas/metabolismo , Mucosa Bucal/efeitos dos fármacos , Extratos Vegetais/farmacologia , Proantocianidinas/farmacologia , Vaccinium macrocarpon/química , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/uso terapêutico , Biofilmes/efeitos dos fármacos , Candida albicans/patogenicidade , Candidíase Bucal/metabolismo , Candidíase Bucal/microbiologia , Adesão Celular/efeitos dos fármacos , Células Epiteliais/efeitos dos fármacos , Interleucina-6/metabolismo , Interleucina-8/metabolismo , Mucosa Bucal/citologia , Mucosa Bucal/microbiologia , NF-kappa B/metabolismo , Fosforilação , Fosfotransferases/metabolismo , Fitoterapia , Extratos Vegetais/uso terapêutico , Proantocianidinas/uso terapêutico , Saliva
19.
PLoS One ; 6(2): e17046, 2011 Feb 23.
Artigo em Inglês | MEDLINE | ID: mdl-21407800

RESUMO

Candida albicans frequently causes superficial infections by invading and damaging epithelial cells, but may also cause systemic infections by penetrating through epithelial barriers. C. albicans is an unusual pathogen because it can invade epithelial cells via two distinct mechanisms: induced endocytosis, analogous to facultative intracellular enteropathogenic bacteria, and active penetration, similar to plant pathogenic fungi. Here we investigated the molecular basis of C. albicans epithelial interactions. By systematically assessing the contributions of defined fungal pathways and factors to different stages of epithelial interactions, we provide an expansive portrait of the processes and activities involved in epithelial infection. We strengthen the concept that hyphal formation is critical for epithelial invasion. Importantly, our data support a model whereby initial epithelial invasion per se does not elicit host damage, but that C. albicans relies on a combination of contact-sensing, directed hyphal extension, active penetration and the expression of novel pathogenicity factors for further inter-epithelial invasion, dissemination and ultimate damage of host cells. Finally, we explore the transcriptional landscape of C. albicans during the early stages of epithelial interaction, and, via genetic analysis, identify ICL1 and PGA34 as novel oral epithelial pathogenicity factors.


Assuntos
Candida albicans/patogenicidade , Células Epiteliais/microbiologia , Células CACO-2 , Candida albicans/genética , Candidíase Bucal/metabolismo , Adesão Celular/fisiologia , Proteínas Fúngicas/metabolismo , Genes Fúngicos/fisiologia , Glioxilatos/metabolismo , Humanos , Isocitrato Liase/metabolismo , Boca , Estresse Fisiológico/genética , Transcrição Gênica/fisiologia , Transcriptoma/fisiologia , Regulação para Cima
20.
Microb Pathog ; 50(6): 278-85, 2011 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-21296654

RESUMO

Candida albicans is a commensal organism that can be isolated from the majority of healthy individuals. However, in certain susceptible individuals C. albicans can become pathogenic leading to the mucocutaneous infection; oral candidiasis. Murine models and in vitro monolayer cultures have generated some data on the likely virulence and host factors that contribute to oral candidiasis but these models have limitations. Recently, tissue engineered oral mucosal models have been developed to mimic the normal oral mucosa but little information is available on their true representation. In this study, we assessed the histological features of three different tissue engineered oral mucosal models compared to the normal oral mucosa and analysed both cell damage and cytokine release following infection with C. albicans. Models comprised of normal oral keratinocytes and a fibroblast-containing matrix displayed more similar immunohistological and proliferation characteristics to normal mucosa, compared to models composed of an oral carcinoma cell line. Although all models were invaded and damaged by C. albicans in a similar manner, the cytokine response was much more pronounced in models containing normal keratinocytes. These data suggest that models based on normal keratinocytes atop a fibroblast-containing connective tissue will significantly aid in dissecting the molecular pathogenesis of oral candidiasis.


Assuntos
Candida albicans/fisiologia , Candidíase Bucal/microbiologia , Candidíase Bucal/patologia , Mucosa Bucal/citologia , Mucosa Bucal/microbiologia , Engenharia Tecidual/métodos , Animais , Candida albicans/genética , Candida albicans/imunologia , Candida albicans/metabolismo , Candidíase Bucal/imunologia , Candidíase Bucal/metabolismo , Morte Celular/fisiologia , Citocinas/biossíntese , Fibroblastos/citologia , Fibroblastos/imunologia , Fibroblastos/metabolismo , Fibroblastos/patologia , Humanos , Imuno-Histoquímica , Queratinócitos/citologia , Queratinócitos/imunologia , Queratinócitos/metabolismo , Queratinócitos/patologia , Camundongos , Mucosa Bucal/imunologia , Mucosa Bucal/metabolismo , Mucosa Bucal/patologia , Regulação para Cima , Virulência/fisiologia , beta-Defensinas/biossíntese , beta-Defensinas/genética
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