title: In vivo and in situ imaging of experimental invasive pulmonary aspergillosis using fibered confocal fluorescence microscopy.
abstract: Invasive pulmonary aspergillosis (IPA) is a highly fatal disease in immunosuppressed patients. In this study, we assessed fibered confocal fluorescence microscopy (FCFM), a new endoscopic technique that enables in vivo microscopic imaging of the distal lung, as a tool for in vivo imaging of IPA. IPA was induced in immunosuppressed rats using a wild strain of Aspergillus fumigatus (n = 6) or a fluorescent transformed TAG-RFP A. fumigatus strain (n = 10). Subpleural areas of pulmonary infection were imaged in vivo using FCFM employing a transthoracic approach. Results were compared to three immunosuppressed control groups, i.e., non-inoculated rats (n = 4), rats inoculated with sterile Phospate-buffer saline (PBS; n = 5), and rats inoculated with Geosmithia argillacea (n = 6). Only hyphae of TAG-RFP A. fumigatus were detectable both in vitro and in vivo by FCFM. In vivo, a local infiltration of fluorescent alveolar macrophages was observed with FCFM in IPA areas in all fungal infections groups, but also in focal inflammatory areas in the immunosuppressed PBS group. A specific fibrillar fluorescence was observed in IPA areas with the TAG-RFP A. fumigatus group, with a 83% sensitivity, a 100% specificity, a 100% positive predictive value and 94% negative predictive value. FCFM provides a new tool to study host-aspergillus interactions in vivo.
authors: Laboratoire LITIS - QuantIF, EA 4108, Rouen University, Rouen, France.
authors: Hélène
authors: H
authors: Morisse
authors: Loraine
authors: L
authors: Heyman
authors: Mathieu
authors: M
authors: Salaün
authors: Loïc
authors: L
authors: Favennec
authors: Jean Michel
authors: JM
authors: Picquenot
authors: Pierre
authors: P
authors: Bohn
authors: Luc
authors: L
authors: Thiberville
mesh_terms: D000818:Animals; D001232:Aspergillus fumigatus / Q000472:pathogenicity; D004195:Disease Models, Animal; D004724:Endoscopy / Q000379:methods*; D007091:Image Processing, Computer-Assisted / Q000379:methods; D055744:Invasive Pulmonary Aspergillosis / Q000175:diagnosis* / Q000473:pathology*; D008168:Lung / Q000473:pathology; D016676:Macrophages, Alveolar / Q000276:immunology; D008297:Male; D018613:Microscopy, Confocal / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D011237:Predictive Value of Tests; D051381:Rats; D017207:Rats, Sprague-Dawley; D012680:Sensitivity and Specificity