Adaptation of a Gaussia princeps Luciferase reporter system in Candida albicans for in vivo detection in the Galleria mellonella infection model

被引:23
作者
Delarze, Eric
Ischer, Francoise
Sanglard, Dominique
Coste, Alix T. [1 ]
机构
[1] Univ Lausanne, Inst Microbiol, Lausanne, Switzerland
基金
瑞士国家科学基金会;
关键词
Candida albicans; Galleria mellonella; in vivo; luciferase; ANTIFUNGAL AGENTS; VIRULENCE; HOST; GENE; PATHOGENICITY; CONSTRUCTION; PATHOGENESIS; EXPRESSION; MUTANTS; LARVAE;
D O I
10.1080/21505594.2015.1081330
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
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.
引用
收藏
页码:684 / 693
页数:10
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