Amphotericin B Resistance in Lomentospora prolificans is associated with a soluble cell wall component

IntroductionLomentospora prolificans is a pathogenic filamentous fungus that causes disease primarily in people with severely compromised immune systems. It is pan-resistant to antifungal drugs, but the mechanism of its resistance to amphotericin B (AMB) is unknown. ObjectivesWe aimed to investigate the mechanism of resistance to AMB of L. prolificans. MethodsThe AMB susceptibility of L. prolificans protoplasts was measured using broth microdilution. L. prolificans, either intact, homogenized or fractionated was incubated with AMB in broth. The same activity was carried out with Aspergillus fumigatus as a control. This broth was then used to prepare microdilution plates with Saccharomyces cerevisiae to determine the activity of the conditioned AMB. ResultsAMB was 16-fold more effective in inhibiting the growth of L. prolificans protoplasts than conidia, but only two-fold more effective against A. fumigatus protoplasts than conidia. Incubation of L. prolificans hyphae with AMB in media diminished drug activity to a much greater extent than A. fumigatus, with 8-fold greater fungal mass of the latter required to achieve the effect of the former. Homogenization and fractionization of L. prolificans revealed that the factor inhibiting AMB activity was soluble with a mass >100 kda. DNase, trypsin, proteinase K, amyloglucosidase, SDS and 0.22 m had no effect on the AMB resistance factor, while treatment with urea, acetonitrile inactivated it. ConclusionWe report a different mechanism for AMB resistance based on the existence of a substance residing in the L. prolificans cell wall that can eliminate the antifungal activity of AMB.