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J Appl Microbiol ; 119(1): 177-87, 2015 Jul.
Article in English | MEDLINE | ID: mdl-25845720

ABSTRACT

AIMS: This study describes a novel in vitro assay that simultaneously determines antifungal efficiency and host cell toxicity using suspensions of human leukaemic cells (HL-60) infected with Candida albicans. METHODS AND RESULTS: The effect of Candida infection on host cell viability was evaluated by the microscopy of trypan blue-stained cells and lactate dehydrogenase (LDH) activity. The in vitro 'drug potency assay' utilized the Cell Counting Kit-8 and measured post-antifungal treatment viability of Candida-infected HL-60 cells and the ability of the antifungal treatment to prevent infection. LDH activity showed that 42% ± 4·0 and 85·3% ± 7·40 of HL-60 cells were killed following Candida infection at the multiplicity of infection (MOI) of 1 : 1 and 1 : 5, respectively. The antifungal nystatin (0·78-25 µmol l(-1) ) was found to inhibit C. albicans infection as seen by the significantly increased viability of HL-60 cells. Cytotoxicity of nystatin towards infected HL-60 cells was evident at higher concentrations and this was also confirmed by propidium iodide staining. CONCLUSIONS: An assay using undisturbed cell suspension conditions was successfully developed for assessing the selectivity of the antifungal therapy in the host-Candida environment. SIGNIFICANCE AND IMPACT OF THE STUDY: 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.


Subject(s)
Antifungal Agents/toxicity , Biological Assay/methods , Candida albicans/drug effects , Candidiasis/microbiology , Nystatin/toxicity , Candida albicans/physiology , Candidiasis/physiopathology , Cell Survival/drug effects , HL-60 Cells , Humans
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