title: Human beta-defensins kill Candida albicans in an energy-dependent and salt-sensitive manner without causing membrane disruption.
abstract: Human beta-defensin 2 (hBD-2) and hBD-3 have potent fungicidal activity in the micromolar range. Although little is known about their mechanism of action against Candida species, some similarities to the antifungal mechanism of salivary peptide histatin 5 (Hst 5) seem to exist. Since hBD-2 and hBD-3 have been reported to cause direct disruption of target cell membranes, we compared the effects of hBD-2 and hBD-3 on Candida albicans membrane integrity. Incubation of calcein-loaded C. albicans cells with a dose of hBD-2 lethal for 90% of the strains tested (LD(90)) resulted in a maximal dye efflux of only 10.3% +/- 2.8% at 90 min, similar to that induced by Hst 5. In contrast, an LD(90) of hBD-3 more than doubled calcein release from cells yet did not result in more than 24% of total release, showing that neither peptide caused gross membrane damage. As for Hst 5, killing of C. albicans cells by hBD-2 and hBD-3 was salt sensitive; however, Ca(2+) and Mg(2+) inhibited hBD-2 but not hBD-3 fungicidal activity. Pretreatment of C. albicans cells with sodium azide resulted in significantly decreased ATP release and susceptibility of cells to hBD-2 and hBD-3. However, hBD-3 killing was partially restored at concentrations of > or =0.8 microM, showing energy-independent mechanisms at higher doses. C. glabrata resistance to Hst 5, hBD-2, and hBD-3 is not a result of loss of expression of cell wall Ssa proteins. The candidacidal effects of hBD-2-hBD-3 and Hst 5-hBD-2 were additive, while the index of interaction between Hst 5 and hBD-3 was 0.717 (P < 0.05). Thus, the candidacidal action of hBD-2 shows many similarities to that of Hst 5 in terms of salt sensitivity, ion selectivity, and energy requirements while hBD-3 exhibits biphasic concentration-dependent mechanisms of candidacidal action complementary to those of Hst 5.
authors.affiliation: Department of Oral Biology, State University of New York at Buffalo, Buffalo, NY 14214, USA.
authors.forename: Slavena
authors.initials: S
authors.lastname: Vylkova
authors.forename: Namrata
authors.initials: N
authors.lastname: Nayyar
authors.forename: Wansheng
authors.initials: W
authors.lastname: Li
authors.forename: Mira
authors.initials: M
authors.lastname: Edgerton
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000935:Antifungal Agents / Q000494:pharmacology; D015153:Blotting, Western; D002122:Calcium Chloride / Q000494:pharmacology; D002176:Candida albicans / Q000187:drug effects*; D002462:Cell Membrane / Q000187:drug effects* / Q000378:metabolism; D002473:Cell Wall / Q000187:drug effects / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D054808:Histatins; D006801:Humans; D010710:Phosphates / Q000494:pharmacology; D012471:Salivary Proteins and Peptides / Q000494:pharmacology; D012965:Sodium Chloride / Q000494:pharmacology; D023083:beta-Defensins / Q000494:pharmacology*
chemical_list: D000935:Antifungal Agents; C422950:DEFB103A protein, human; C068075:HTN3 protein, human; D054808:Histatins; D010710:Phosphates; D012471:Salivary Proteins and Peptides; D023083:beta-Defensins; D012965:Sodium Chloride; D000255:Adenosine Triphosphate; D002122:Calcium Chloride; C018279:sodium phosphate