This Article Full Text (PDF) Other Versions of this Article: AEM.01288-08v1 74/22/7023    most recent Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Google Scholar Articles by Brock, M. Articles by Ibrahim-Granet, O. PubMed PubMed Citation Articles by Brock, M. Articles by Ibrahim-Granet, O. Agricola Articles by Brock, M. Articles by Ibrahim-Granet, O.

 Previous Article  |  Next Article 

Appl. Environ. Microbiol. doi:10.1128/AEM.01288-08
Copyright (c) 2008, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Bioluminescent Aspergillus fumigatus: A new tool for drug efficiency testing and in vivo monitoring of invasive aspergillosis

Leibniz Institute for Natural Product Research and Infection Biology, –Hans Knoell Institute–, Junior Research Group Microbial Biochemistry and Physiology, Beutenbergstr. 11a, 07745 Jena, Germany; Unité de Recherche et d'Expertise Histotechnologie et Pathologie, Institut Pasteur, 25 rue du Dr Roux, 75724 Paris cedex 15, France; Unité des Interactions Bactéries-Cellules, Institut Pasteur, 25 rue du Dr Roux, 75724 Paris cedex 15, France; Plate-Forme d'Imagerie Dynamique, Institut Pasteur, 25 rue du Dr Roux, 75724 Paris cedex 15, France; Unité Cytokines & Inflammation, Institut Pasteur, 28 rue du Dr Roux, 75724 Paris cedex 15, France

^* To whom correspondence should be addressed. Email: Matthias.brock{at}hki-jena.de.

	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.