“…While IL-8 is a known neutrophil recruiter from the circulating blood to the site of infection ( Höffs et al, 2016 ), IL-6 is related to immune response to microorganisms and its expression is directly influenced by the secretion of other pro-inflammatory cytokines, especially IL-1 ( Tanaka et al, 2014 ). Some reports state that C. albicans infection is crucial for the modulation and release of IL-1α, IL-6, and IL-8, promoting a higher production of these cytokines ( Feller et al, 2014 ; Figueira et al, 2020 ; Mo et al, 2020 ). In this regard, our findings suggest that A. colubrina might reduce pro-inflammatory IL-6 and IL-8 cytokines expression during the fungal infection, due its strong antifungal activity, which modulates some putative virulence factors of the C. albicans , such as biofilm formation and proteolytic enzyme secretion, reducing the fungus pathogenicity.…”
Oral candidiasis is one of the most common fungal infections in humans. Its incidence has increased widely, as well as the antifungal resistance, demanding for the search for novel antifungal therapeutic agents. Anadenanthera colubrina (Vell.) Brenan is a plant species that has been proven to possess pharmacological effects, including antifungal and anti-inflammatory activities. This study evaluated in vitro the effects of standardized A. colubrina extract on virulence factors of Candida albicans and its regulation on immune response through C. albicans-host interaction. Antifungal activity was evaluated by Broth Microdilution Method against reference Candida strains (C. albicans, C. glabrata, C. tropicalis; C. dubliniensis). Anti-biofilm effect was performed on C. albicans mature biofilm and quantified by CFU/mL/g of biofilm dry weight. Proleotlytic enzymatic activities of proteinase and phospholipase were assessed by Azocasein and Phosphatidylcholine assays, respectively. Cytotoxicity effect was determined by Cell Titer Blue Viability Assay on Human Gingival Fibroblasts. Co-cultured model was used to analyze C. albicans coexisting with HGF by Scanning Electron Microscopy and fluorescence microscopies; gene expression was assessed by RT-PCR of C. albicans enzymes (SAP-1, PLB-1) and of host inflammatory cytokines (IL-6, IL-8, IL-1β, IL-10). Cytokines secretion was analysed by Luminex. The extract presented antifungal effect with MIC<15.62 μg/ml against Candida strains. Biofilm and proteolytic activity were significant reduced at 312.4 μg/ml (20 × 15.62 μg/ml) extract concentration. Cell viability was maintained higher than 70% in concentrations up to 250 μg/ml (LD50 = 423.3 μg/ml). Co-culture microscopies demonstrated a substantial decreased in C. albicans growth and minimal toxicity against host cells. Gene expressions of SAP-1/PLB-1 were significantly down-regulated and host immune response was modulated by a significant decreased on IL-6 and IL-8 cytokines secretion. A. colubrina had antifungal activity on Candida strains, antibiofilm, and anti-proteolytic enzyme effects against C. albicans. Presented low cytotoxicity to the host cells and modulatory effects on the host immune response.
“…While IL-8 is a known neutrophil recruiter from the circulating blood to the site of infection ( Höffs et al, 2016 ), IL-6 is related to immune response to microorganisms and its expression is directly influenced by the secretion of other pro-inflammatory cytokines, especially IL-1 ( Tanaka et al, 2014 ). Some reports state that C. albicans infection is crucial for the modulation and release of IL-1α, IL-6, and IL-8, promoting a higher production of these cytokines ( Feller et al, 2014 ; Figueira et al, 2020 ; Mo et al, 2020 ). In this regard, our findings suggest that A. colubrina might reduce pro-inflammatory IL-6 and IL-8 cytokines expression during the fungal infection, due its strong antifungal activity, which modulates some putative virulence factors of the C. albicans , such as biofilm formation and proteolytic enzyme secretion, reducing the fungus pathogenicity.…”
Oral candidiasis is one of the most common fungal infections in humans. Its incidence has increased widely, as well as the antifungal resistance, demanding for the search for novel antifungal therapeutic agents. Anadenanthera colubrina (Vell.) Brenan is a plant species that has been proven to possess pharmacological effects, including antifungal and anti-inflammatory activities. This study evaluated in vitro the effects of standardized A. colubrina extract on virulence factors of Candida albicans and its regulation on immune response through C. albicans-host interaction. Antifungal activity was evaluated by Broth Microdilution Method against reference Candida strains (C. albicans, C. glabrata, C. tropicalis; C. dubliniensis). Anti-biofilm effect was performed on C. albicans mature biofilm and quantified by CFU/mL/g of biofilm dry weight. Proleotlytic enzymatic activities of proteinase and phospholipase were assessed by Azocasein and Phosphatidylcholine assays, respectively. Cytotoxicity effect was determined by Cell Titer Blue Viability Assay on Human Gingival Fibroblasts. Co-cultured model was used to analyze C. albicans coexisting with HGF by Scanning Electron Microscopy and fluorescence microscopies; gene expression was assessed by RT-PCR of C. albicans enzymes (SAP-1, PLB-1) and of host inflammatory cytokines (IL-6, IL-8, IL-1β, IL-10). Cytokines secretion was analysed by Luminex. The extract presented antifungal effect with MIC<15.62 μg/ml against Candida strains. Biofilm and proteolytic activity were significant reduced at 312.4 μg/ml (20 × 15.62 μg/ml) extract concentration. Cell viability was maintained higher than 70% in concentrations up to 250 μg/ml (LD50 = 423.3 μg/ml). Co-culture microscopies demonstrated a substantial decreased in C. albicans growth and minimal toxicity against host cells. Gene expressions of SAP-1/PLB-1 were significantly down-regulated and host immune response was modulated by a significant decreased on IL-6 and IL-8 cytokines secretion. A. colubrina had antifungal activity on Candida strains, antibiofilm, and anti-proteolytic enzyme effects against C. albicans. Presented low cytotoxicity to the host cells and modulatory effects on the host immune response.
“…Cytokines in the CVL were quantified by ELISA assay using TNF-α, IL-6, IL-1β, TGFβ, IL-22, IL-23, and IL-17 detection kits (R&D System, McKinley Place, MN, USA) [19]. The samples were previously concentrated with a Vivaspin-6 centrifugal concentrator (GE Healthcare, Chicago, IL, USA).…”
Section: Cytokine Immunoassaysmentioning
confidence: 99%
“…It has been reported that recombinant mBD1, a murine homolog of hBD1 [14], can display direct fungicidal activity [15]. BD regulation has been well documented in Candida infection, especially in the gastrointestinal tract, and also in the course of human esophagitis [16], oropharyngeal and gastric murine models [17,18], in vitro studies [17,19], and in human oral infections with reduced levels of salivary hDB1-3 [20]. In a murine model of C. albicans oral infection, Tomalka et al [21] reported that, at early times postinfection (D3 pi), mBD1 deficient mice exhibit 10-fold higher CFU than WT animals and lower PMNs infiltration in the mucosa.…”
Vulvovaginal candidiasis (VVC) and recurrent vulvovaginal candidiasis (RVVC) are two forms of a disease caused by Candida spp. β-defensin (BD) is one of the most important families of antimicrobial peptides in the female genital tract and includes molecules that exert essential local functions as antimicrobial and PMN chemoattractant peptides. However, the information on their role during murine and human VVC and RVVC is limited. Thus, we analyzed the behavior and contribution of BD1 to the local response in a VVC mice model and the local cytokine profile and human BD1 and BD3 expression in cervicovaginal lavage from patients with VVC and RVVC. We demonstrated that, in patients with RVVC BD1, mRNA and protein expression were severely diminished and that the aspartate proteinase and lipase secreted by C. albicans are involved in that decrease. This study provides novel information about the pathogenesis of VVC and describes a highly efficient C. albicans escape strategy for perpetuating the infection; these results may contribute to the development of new or combined treatment approaches.
“…GAPDH, as a kind of moonlighting protein, not only plays a role in the process of glucose metabolism, but also has a certain regulatory effect on the pathogenic capacity of bacteria (Giménez et al, 2014). More recent research shows that the glucose effect on Candida albicans biofilm during tissue invasion (Figueira et al, 2020). Therefore, in this study, we aimed to investigate whether GAPDH, a key regulator of glucose metabolism, affects biofilm formation in B. cereus 0-9.…”
Bacillus cereus 0-9, a Gram-positive endospore-forming bacterium isolated from healthy wheat roots, has biological control capacity against several soil-borne plant diseases of wheat such as sharp eyespot and take-all. The bacterium can produce various biofilms that differ in their architecture and formation mechanisms, possibly for adapting to different environments. The gapB gene, encoding a glyceraldehyde-3-phosphate dehydrogenase (GAPDH), plays a key role in B. cereus 0-9 biofilm formation. We studied the function of GapB and the mechanism of its involvement in regulating B. cereus 0-9 biofilm formation. GapB has GAPDH activities for both NAD+- and NADP+-dependent dehydrogenases and is a key enzyme in gluconeogenesis. Biofilm yield of the ΔgapB strain decreased by 78.5% compared with that of wild-type B. cereus 0-9 in lysogeny broth supplemented with some mineral salts (LBS), and the ΔgapB::gapB mutants were recovered with gapB gene supplementation. Interestingly, supplementing the LBS medium with 0.1–0.5% glycerol restored the biofilm formation capacity of the ΔgapB mutants. Therefore, GapB regulates biofilm formation relative to its function in gluconeogenesis. To illustrate how GapB is involved in regulating biofilm formation through gluconeogenesis, we carried out further research. The results indicate that the GapB regulated the B. cereus 0-9 biofilm formation independently of the exopolysaccharides and regulatory proteins in the typical SinI/R system, likely owing to the release of extracellular DNA in the matrix. Transcriptome analysis showed that the gapB deletion caused changes in the expression levels of only 18 genes, among which, lrgAB was the most significantly increased by 6.17-fold. We confirmed this hypothesis by counting the dead and living cells in the biofilms and found the number of living cells in the biofilm formed by the ΔgapB strain was nearly 7.5 times than that of wild-type B. cereus 0-9. Therefore, we concluded that the GapB is involved in the extracellular DNA release and biofilm formation by regulating the expression or activities of LrgAB. These results provide a new insight into the regulatory mechanism of bacterial biofilm formation and a new foundation for further studying the stress resistance of B. cereus.
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