title: Adaptation of a Gaussia princeps Luciferase reporter system in Candida albicans for in vivo detection in the Galleria mellonella infection model.
abstract: For the past 10 years, mini-host models and in particular the greater wax moth Galleria mellonella have tended to become a surrogate for murine models of fungal infection mainly due to cost, ethical constraints and ease of use. Thus, methods to better assess the fungal pathogenesis in G. mellonella need to be developed. In this study, we implemented the detection of Candida albicans cells expressing the Gaussia princeps luciferase in its cell wall in infected larvae of G. mellonella. We demonstrated that detection and quantification of luminescence in the pulp of infected larvae is a reliable method to perform drug efficacy and C. albicans virulence assays as compared to fungal burden assay. Since the linearity of the bioluminescent signal, as compared to the CFU counts, has a correlation of R(2) = 0.62 and that this method is twice faster and less labor intensive than classical fungal burden assays, it could be applied to large scale studies. We next visualized and followed C. albicans infection in living G. mellonella larvae using a non-toxic and water-soluble coelenterazine formulation and a CCD camera that is commonly used for chemoluminescence signal detection. This work allowed us to follow for the first time C. albicans course of infection in G. mellonella during 4 days.
authors: a Institute of Microbiology; University of Lausanne and University Hospital Center ; Lausanne , Switzerland.
authors: Eric
authors: E
authors: Delarze
authors: a Institute of Microbiology; University of Lausanne and University Hospital Center ; Lausanne , Switzerland.
authors: Françoise
authors: F
authors: Ischer
authors: a Institute of Microbiology; University of Lausanne and University Hospital Center ; Lausanne , Switzerland.
authors: Dominique
authors: D
authors: Sanglard
authors: a Institute of Microbiology; University of Lausanne and University Hospital Center ; Lausanne , Switzerland.
authors: Alix T
authors: AT
authors: Coste
mesh_terms: D000222:Adaptation, Physiological; D000818:Animals; D002176:Candida albicans / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D002177:Candidiasis / Q000382:microbiology; D033342:Copepoda / Q000201:enzymology* / Q000235:genetics; D004195:Disease Models, Animal; D015725:Fluconazole / Q000494:pharmacology; D007093:Imidazoles / Q000494:pharmacology; D007814:Larva / Q000382:microbiology; D007915:Lepidoptera / Q000235:genetics / Q000378:metabolism / Q000382:microbiology*; D008156:Luciferases / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics; D008163:Luminescent Measurements / Q000379:methods; D009036:Moths / Q000382:microbiology; D011719:Pyrazines / Q000494:pharmacology; D014774:Virulence
chemical_list: D007093:Imidazoles; D011719:Pyrazines; C017144:coelenterazine; D015725:Fluconazole; D008156:Luciferases
keywords: Candida albicans; Galleria mellonella; in vivo; luciferase