title: Reduced azole susceptibility of oral isolates of Candida albicans from HIV-positive patients and a derivative exhibiting colony morphology variation.
abstract: Approximately 50% (15/28) of a selection of oral isolates of Candida albicans from separate individuals infected with the human immunodeficiency virus (HIV) exhibited low susceptibility to ketoconazole as determined by hyphal elongation assessment. Nine of these isolates exhibited colony morphology variation or switching at 37 degrees C, of which six expressed low ketoconazole susceptibility. To determine whether colony morphology variation could give rise to derivatives with reduced azole susceptibility, several high-frequency switching variants of three HIV-patient isolates were recovered and assessed. All but one of the variants expressed similar azole susceptibility profiles to their respective parental strains. However, the C. albicans derivative 132ACR expressed significantly reduced susceptibility to ketoconazole in comparison to its parental strain 132A. In whole cells, on the basis of total growth the switched derivative 132ACR was markedly less susceptible than its parental isolate 132A to ketoconazole at 10 microM. A much smaller difference was observed with fluconazole at 10 microM, with the switched derivative 132ACR exhibiting a threefold lower susceptibility compared with the parental isolate 132A. The incorporation of [14C]acetate in control and azole-treated cells of both organisms was higher for the parental strain. When cell lysates of strain 132A and its derivative 132ACR were incubated with [14C]mevalonic acid and ketoconazole, the IC50 for 14C-label incorporation into C-4 demethyl sterols was fivefold higher for lysates of the switched derivative 132ACR compared with those of the parental strain 132A. With fluconazole the IC50 value for the derivative 132ACR was 25-fold higher than for strain 132A. The 14-sterol demethylase of the switched derivative 132ACR was possibly less sensitive to azole inhibition than that of the enzyme of strain 132A. These studies indicated that colony morphology variation in vitro can generate derivatives with stable, reduced azole susceptibility without prior exposure to azoles.
authors.affiliation: Trinity College, Department of Pharmacology, School of Pharmacy, University of Dublin, Republic of Ireland.
authors.forename: P J
authors.initials: PJ
authors.lastname: Gallagher
authors.forename: D E
authors.initials: DE
authors.lastname: Bennett
authors.forename: M C
authors.initials: MC
authors.lastname: Henman
authors.forename: R J
authors.initials: RJ
authors.lastname: Russell
authors.forename: S R
authors.initials: SR
authors.lastname: Flint
authors.forename: D B
authors.initials: DB
authors.lastname: Shanley
authors.forename: D C
authors.initials: DC
authors.lastname: Coleman
mesh_terms: D002176:Candida albicans / Q000187:drug effects* / Q000254:growth & development; D002180:Candidiasis, Oral / Q000382:microbiology; D004271:DNA, Fungal / Q000235:genetics; D015725:Fluconazole / Q000494:pharmacology*; D015658:HIV Infections / Q000382:microbiology; D006801:Humans; D007654:Ketoconazole / Q000494:pharmacology*; D008055:Lipids / Q000096:biosynthesis; D008826:Microbial Sensitivity Tests; D013261:Sterols / Q000096:biosynthesis
chemical_list: D004271:DNA, Fungal; D008055:Lipids; D013261:Sterols; D015725:Fluconazole; D007654:Ketoconazole