A novel <i>in vitro</i>
 assay for assessing efficacy and toxicity of antifungals using human leukaemic cells infected with <i>Candida albicans</i>

Boros-Majewska, Joanna; Turczyk, Lukasz; Wei, Xiao-Qing; Milewski, Sławomir; Williams, David Wynne

doi:10.1111/jam.12817

Cited by 6 publications

(4 citation statements)

References 42 publications

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“…Confocal laser scanning LIVE/DEAD â staining of RHE RHE samples from each time point were placed on microscope slides and 100 ll of LIVE/DEAD â stain (containing 25 lmol l À1 SYTO â 9 and 15 lmol l À1 propidium iodide) applied directly to the tissue, as previously described (Boros-Majewska et al 2015). Treated samples were incubated for 30 min at 37°C in the dark prior to transfer to clean glass slides.…”

Section: Hyphal Invasion Assaymentioning

confidence: 99%

Alginate oligosaccharides modify hyphal infiltration ofCandida albicansin anin vitromodel of invasive human candidosis

et al. 2017

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Section: Hyphal Invasion Assaymentioning

confidence: 99%

Alginate oligosaccharides modify hyphal infiltration ofCandida albicansin anin vitromodel of invasive human candidosis

et al. 2017

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“…Furthermore, it was shown to be higher than the uninfected control. 22 There was a significant difference in the number of inflammatory cells between the negative and positive control groups. The significant difference between the results of the holothurin treatment given directly to the Wistar vagina revealed that it was lower than the administration of caspofungin.…”

Section: Discussionmentioning

confidence: 91%

The Effects of Holothurin and Caspofungin on the Vaginal Cell Inflammation Parameters of the Rattus norvegicus Strain Post Induction of Candida albicans

Handayani¹,

Nurdiana²,

Winarsih³

et al. 2023

IDR

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Candida albicans (C. albicans) is a fungus that causes superficial and invasive candidiasis in its host. Caspofungin, has been widely used as a synthetic antifungal, whereas holothurin has been shown to have potential as a natural antifungal. The purpose of this study was to see how holothurin and caspofungin affected the number of C. albicans's colonies, LDH levels, and the number of inflammatory cells in vagina of Rattus norvegicus. Patients and Methods: Design of this research is using posttest only with control group design with 48 Rattus norvegicus Wistar strains used in this study were divided into six treatment groups. Each group was divided into three-time intervals of 12, 24, and 48 hours. LDH markers were tested using ELISA, inflammatory cells were counted manually, and the number of colonies was calculated using colonymetry before being diluted with NaCl 0.9% and planted in sabouraud dextrose agar (SDA). Results: According to the findings, inflammatory cells in the treatment of holothurin (48-hours) had an OR of 1.68 CI (−0.79-4.16) P = 0.09 and caspofungin had an OR of 4.18 CI (1.26-9.63) P = 0.09. Meanwhile, LDH in the holothurin (48-hour) treatment obtained OR 348, CI (286-410), P=0.03, and Caspofungin OR 393, CI (277-508), P=0.03. Colonies were obtained with zero numbers in the holothurin treatment (48 hours) and with Caspofungin OR 393, CI (273-508) P=0.00. Conclusion: Holothurin and caspofungin administration reduced the number of C. albicans colonies and the number of inflammatory cells (P 0.05), implying that holothurin and caspofungin could prevent C. albicans infection.

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“…The viability of neutrophils upon exposure to yeasts is an important indication on the type of NETosis, lytic or non-lytic ( Yang et al., 2016 ; Papayannopoulos, 2018 ). In this work, we quantified the neutrophils’ viability using a trypan blue exclusion assay, and although there are other quantification methodologies considered more reliable and sensitive ( Hosseinzadeh et al, 2012 ; Boros-Majewska et al., 2015 ), the robustness and consistency of the results justifies its validity. This observation reinforces the idea that yeasts induce a non-lytic NETosis, at least in the cases of C. albicans and C. glabrata .…”

Section: Discussionmentioning

confidence: 99%

Candida Extracellular Nucleotide Metabolism Promotes Neutrophils Extracellular Traps Escape

Afonso

Mestre

Silva

et al. 2021

Front. Cell. Infect. Microbiol.

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Host innate immunity is fundamental to the resistance against Candida albicans and Candida glabrata infection, two of the most important agents contributing to human fungal infections. Phagocytic cells, such as neutrophils, constitute the first line of host defense mechanisms, and the release of neutrophil extracellular traps (NETs) represent an important strategy to immobilize and to kill invading microorganisms, arresting the establishment of infection. The purinergic system operates an important role in the homeostasis of immunity and inflammation, and ectophosphatase and ectonucleotidase activities are recognized as essential for survival strategies and infectious potential of several pathogens. The expression and unique activity of a 3′-nucleotidase/nuclease (3′NT/NU), able to hydrolyze not only AMP but also nucleic acids, has been considered as part of a possible mechanism of microbes to escape from NETs. The aim of the present study was to evaluate if yeasts escape from the NET-mediated killing through their 3′NT/NU enzymatic activity contributing to NET-hydrolysis. After demonstrating the presence of 3′NT/NU activity in C. albicans, C. glabrata, and Saccharomyces cerevisiae, we show that, during neutrophils-Candida interaction, when NETs formation and release are triggered, NETs digestion occurs and this process of NETs disruption promoted by yeast cells was prevented by ammonium tetrathiomolybdate (TTM), a 3′NT/NU inhibitor. In conclusion, although the exact nature and specificity of yeasts ectonucleotidases are not completely unraveled, we highlight the importance of these enzymes in the context of infection, helping yeasts to overcome host defenses, whereby C. albicans and C. glabrata can escape NET-mediate killing through their 3′NT/NU activity.

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A novel in vitro assay for assessing efficacy and toxicity of antifungals using human leukaemic cells infected with Candida albicans

Abstract: The assay employing Candida infection of host cell suspensions represents a promising method for testing interactions of antifungal compounds with both fungal and host cells.

Cited by 6 publications

References 42 publications

Alginate oligosaccharides modify hyphal infiltration ofCandida albicansin anin vitromodel of invasive human candidosis

Alginate oligosaccharides modify hyphal infiltration ofCandida albicansin anin vitromodel of invasive human candidosis

The Effects of Holothurin and Caspofungin on the Vaginal Cell Inflammation Parameters of the Rattus norvegicus Strain Post Induction of Candida albicans

Candida Extracellular Nucleotide Metabolism Promotes Neutrophils Extracellular Traps Escape

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