title: Gliotoxin is a virulence factor of Aspergillus fumigatus: gliP deletion attenuates virulence in mice immunosuppressed with hydrocortisone.
abstract: Gliotoxin is an immunosuppressive mycotoxin long suspected to be a potential virulence factor of Aspergillus fumigatus. Recent studies using mutants lacking gliotoxin production, however, suggested that the mycotoxin is not important for pathogenesis of A. fumigatus in neutropenic mice resulting from treatment with cyclophosphomide and hydrocortisone. In this study, we report on the pathobiological role of gliotoxin in two different mouse strains, 129/Sv and BALB/c, that were immunosuppressed by hydrocortisone alone to avoid neutropenia. These strains of mice were infected using the isogenic set of a wild type strain (B-5233) and its mutant strain (gliPDelta) and the the glip reconstituted strain (gliP(R)). The gliP gene encodes a nonribosomal peptide synthase that catalyzes the first step in gliotoxin biosynthesis. The gliPDelta strain was significantly less virulent than strain B-5233 or gliP(R) in both mouse models. In vitro assays with culture filtrates (CFs) of B-5233, gliPDelta, and gliP(R) strains showed the following: (i) deletion of gliP abrogated gliotoxin production, as determined by high-performance liquid chromatography analysis; (ii) unlike the CFs from strains B-5233 and gliP(R), gliPDelta CFs failed to induce proapoptotic processes in EL4 thymoma cells, as tested by Bak conformational change, mitochondrial-membrane potential disruption, superoxide production, caspase 3 activation, and phosphatidylserine translocation. Furthermore, superoxide production in human neutrophils was strongly inhibited by CFs from strain B-5233 and the gliP(R) strain, but not the gliPDelta strain. Our study confirms that gliotoxin is an important virulence determinant of A. fumigatus and that the type of immunosuppression regimen used is important to reveal the pathogenic potential of gliotoxin.
authors.affiliation: Laboratory of Clinical Infectious Diseases, National Institute of Allergy and Infectious Diseases, NIH, Bethesda, MD 20892, USA.
authors.forename: Janyce A
authors.initials: JA
authors.lastname: Sugui
authors.forename: Julian
authors.initials: J
authors.lastname: Pardo
authors.forename: Yun C
authors.initials: YC
authors.lastname: Chang
authors.forename: Kol A
authors.initials: KA
authors.lastname: Zarember
authors.forename: Glenn
authors.initials: G
authors.lastname: Nardone
authors.forename: Eva M
authors.initials: EM
authors.lastname: Galvez
authors.forename: Arno
authors.initials: A
authors.lastname: Müllbacher
authors.forename: John I
authors.initials: JI
authors.lastname: Gallin
authors.forename: Markus M
authors.initials: MM
authors.lastname: Simon
authors.forename: Kyung J
authors.initials: KJ
authors.lastname: Kwon-Chung
mesh_terms: D000818:Animals; D017209:Apoptosis; D001228:Aspergillosis / Q000382:microbiology* / Q000473:pathology; D001232:Aspergillus fumigatus / Q000235:genetics / Q000472:pathogenicity*; D005656:Fungal Proteins / Q000235:genetics*; D017353:Gene Deletion; D005912:Gliotoxin / Q000096:biosynthesis*; D006801:Humans; D006854:Hydrocortisone / Q000494:pharmacology; D007165:Immunosuppression Therapy; D051379:Mice; D008807:Mice, Inbred BALB C; D009504:Neutrophils / Q000378:metabolism / Q000382:microbiology; D010453:Peptide Synthases / Q000235:genetics*; D016897:Respiratory Burst; D013481:Superoxides / Q000378:metabolism; D014774:Virulence / Q000235:genetics; D037521:Virulence Factors / Q000096:biosynthesis* / Q000235:genetics; D050998:bcl-2 Homologous Antagonist-Killer Protein / Q000378:metabolism
chemical_list: D005656:Fungal Proteins; D037521:Virulence Factors; D050998:bcl-2 Homologous Antagonist-Killer Protein; D013481:Superoxides; D005912:Gliotoxin; D010453:Peptide Synthases; C104385:non-ribosomal peptide synthase; D006854:Hydrocortisone