title: Disruption of the phospholipase D gene attenuates the virulence of Aspergillus fumigatus.
abstract: Aspergillus fumigatus is the most prevalent airborne fungal pathogen that induces serious infections in immunocompromised patients. Phospholipases are key enzymes in pathogenic fungi that cleave host phospholipids, resulting in membrane destabilization and host cell penetration. However, knowledge of the impact of phospholipases on A. fumigatus virulence is rather limited. In this study, disruption of the pld gene encoding phospholipase D (PLD), an important member of the phospholipase protein family in A. fumigatus, was confirmed to significantly decrease both intracellular and extracellular PLD activity of A. fumigatus. The pld gene disruption did not alter conidial morphological characteristics, germination, growth, and biofilm formation but significantly suppressed the internalization of A. fumigatus into A549 epithelial cells without affecting conidial adhesion to epithelial cells. Importantly, the suppressed internalization was fully rescued in the presence of 100 μM phosphatidic acid, the PLD product. Indeed, complementation of pld restored the PLD activity and internalization capacity of A. fumigatus. Phagocytosis of A. fumigatus conidia by J774 macrophages was not affected by the absence of the pld gene. Pretreatment of conidia with 1-butanol and a specific PLD inhibitor decreased the internalization of A. fumigatus into A549 epithelial cells but had no effect on phagocytosis by J774 macrophages. Finally, loss of the pld gene attenuated the virulence of A. fumigatus in mice immunosuppressed with hydrocortisone acetate but not with cyclophosphamide. These data suggest that PLD of A. fumigatus regulates its internalization into lung epithelial cells and may represent an important virulence factor for A. fumigatus infection.
authors.affiliation: Department of Immunology, Medical College of Qingdao University, Qingdao, China.
authors.forename: Xianping
authors.initials: X
authors.lastname: Li
authors.forename: Meihua
authors.initials: M
authors.lastname: Gao
authors.forename: Xuelin
authors.initials: X
authors.lastname: Han
authors.forename: Sha
authors.initials: S
authors.lastname: Tao
authors.forename: Dongyu
authors.initials: D
authors.lastname: Zheng
authors.forename: Ying
authors.initials: Y
authors.lastname: Cheng
authors.forename: Rentao
authors.initials: R
authors.lastname: Yu
authors.forename: Gaige
authors.initials: G
authors.lastname: Han
authors.forename: Martina
authors.initials: M
authors.lastname: Schmidt
authors.forename: Li
authors.initials: L
authors.lastname: Han
mesh_terms: D000818:Animals; D001232:Aspergillus fumigatus / Q000201:enzymology* / Q000254:growth & development / Q000472:pathogenicity* / Q000502:physiology; D018441:Biofilms / Q000254:growth & development; D002448:Cell Adhesion; D002460:Cell Line; D004847:Epithelial Cells / Q000382:microbiology; D017353:Gene Deletion; D005816:Genetic Complementation Test; D006801:Humans; D008264:Macrophages / Q000276:immunology / Q000382:microbiology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D010587:Phagocytosis; D010739:Phospholipase D / Q000235:genetics / Q000378:metabolism*; D013172:Spores, Fungal / Q000472:pathogenicity; D014774:Virulence; D037521:Virulence Factors / Q000235:genetics / Q000378:metabolism*
chemical_list: D037521:Virulence Factors; D010739:Phospholipase D
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