Bioluminescent Aspergillus fumigatus: A new tool for drug efficiency testing and in vivo monitoring of invasive aspergillosis
Author:
Brock M, Jouvion G, Droin-BergÈre S, Dussurget O, Nicola MA, Ibrahim-Granet O
Date: 27 October 2008
Abstract:
Aspergillus fumigatus is the main cause of invasive aspergillosis in immunocompromised patients and only a limited number of drugs for treatment are available. A screening method for new antifungal compounds is urgently required, preferably an approach suitable for in vitro and in vivo studies. Bioluminescence imaging is a powerful tool to study the temporal and spatial resolution of the infection and the effectiveness of antifungal drugs. Here, we describe the construction of a bioluminescent A. fumigatus strain by fusing the promoter of the glyceraldehyde-3-phosphate dehydrogenase from A. fumigatus with the luciferase gene from Photinus pyralis to control the expression of the bioluminescent reporter. A. fumigatus transformed with this construct revealed high bioluminescence under all tested growth conditions. Furthermore, light emission correlated with the amount of conidia used for inoculation and with the biomass formed after different incubation times. The bioluminescent strains were suitable to study the effectiveness of antifungals in vitro by several independent methods, including the determination of light emission in a microplate reader and the direct visualisation of light emission in an IVIS 100 system. Moreover, when glucocorticoid treated immunosuppressed mice were infected with a bioluminescent strain, light emission was detected from infected lungs, allowing the visualisation of the progression of invasive aspergillosis. Therefore, this new bioluminescent tool is suitable to study the in vitro effectiveness of drugs, the disease development, localisation and burden of fungi within tissues and may also provide a powerful tool to study the effectiveness of antifungals in vivo.
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