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

• Published in: Journal of applied microbiology Vol. 119; no. 1; pp. 177 - 187
• Main Authors: Boros‐Majewska, J., Turczyk, Ł., Wei, X., Milewski, S., Williams, D.W.
• Format: Journal Article
• Language: English
• Published: England Oxford University Press 01.07.2015
• Subjects: Antifungal Agents - toxicity; antimicrobials; Bioassays; Biological Assay - methods; Candida albicans; Candida albicans - drug effects; Candida albicans - physiology; Candidiasis - microbiology; Candidiasis - physiopathology; Cell Survival - drug effects; Fungal infections; fungi; HL-60 Cells; Humans; infection; Leukemia; Microbiology; mycology; nystatin; Nystatin - toxicity; Toxicity; infection; antimicrobials; fungi; nystatin; mycology
• ISSN: 1364-5072; 1365-2672
• DOI: 10.1111/jam.12817

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.