title: Macrophage reporter cell assay for screening immunopharmacological activity of cell wall-active antifungals.
abstract: Antifungal exposure can elicit immunological effects that contribute to activity in vivo, but this activity is rarely screened in vitro in a fashion analogous to MIC testing. We used RAW 264.7 murine macrophages that express a secreted embryonic alkaline phosphatase (SEAP) gene induced by transcriptional activation of NF-κB and activator protein 1 (AP-1) to develop a screen for immunopharmacological activity of cell wall-active antifungal agents. Isolates of Candida albicans and Aspergillus fumigatus that conditionally express genes involved in cell wall synthesis were also tested with the reporter macrophages. We found that growth of fungi in subinhibitory concentrations of glucan synthesis inhibitors (caspofungin and enfumafungin A) or repression of the β-glucan catalytic subunit of glucan synthase, FKS1, increased macrophage NF-κB/AP-1 activation in a dectin-1-dependent manner. This pattern of activation was also transiently observed with repression of chitin synthesis in C. albicans or when yeast cells were incubated in low concentrations of the chitin synthesis inhibitor nikkomycin Z.
authors.affiliation: University of Houston College of Pharmacy, Houston, Texas, USA.
authors.forename: Russell E
authors.initials: RE
authors.lastname: Lewis
authors.forename: Guangling
authors.initials: G
authors.lastname: Liao
authors.forename: Katherine
authors.initials: K
authors.lastname: Young
authors.forename: Cameron
authors.initials: C
authors.lastname: Douglas
authors.forename: Dimitrios P
authors.initials: DP
authors.lastname: Kontoyiannis
mesh_terms: D000469:Alkaline Phosphatase / Q000096:biosynthesis; D000617:Aminoglycosides / Q000494:pharmacology; D000818:Animals; D000935:Antifungal Agents / Q000494:pharmacology*; D001232:Aspergillus fumigatus / Q000187:drug effects* / Q000235:genetics / Q000276:immunology; D002176:Candida albicans / Q000187:drug effects* / Q000235:genetics / Q000276:immunology; D000077336:Caspofungin; D002473:Cell Wall / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D054714:Echinocandins / Q000494:pharmacology; D006027:Glycosides / Q000494:pharmacology; D055666:Lipopeptides; D008264:Macrophages / Q000187:drug effects* / Q000276:immunology; D051379:Mice; D008826:Microbial Sensitivity Tests / Q000379:methods*; D016328:NF-kappa B / Q000096:biosynthesis; D018808:Transcription Factor AP-1 / Q000096:biosynthesis; D014315:Triterpenes / Q000494:pharmacology
chemical_list: D000617:Aminoglycosides; D000935:Antifungal Agents; D054714:Echinocandins; D006027:Glycosides; D055666:Lipopeptides; D016328:NF-kappa B; D018808:Transcription Factor AP-1; D014315:Triterpenes; C421868:enfumafungin; C011952:nikkomycin; D000469:Alkaline Phosphatase; D000077336:Caspofungin