title: Distinct roles for Dectin-1 and TLR4 in the pathogenesis of Aspergillus fumigatus keratitis.
abstract: Aspergillus species are a major worldwide cause of corneal ulcers, resulting in visual impairment and blindness in immunocompetent individuals. To enhance our understanding of the pathogenesis of Aspergillus keratitis, we developed a murine model in which red fluorescent protein (RFP)-expressing A. fumigatus (Af293.1RFP) conidia are injected into the corneal stroma, and disease progression and fungal survival are tracked over time. Using Mafia mice in which c-fms expressing macrophages and dendritic cells can be induced to undergo apoptosis, we demonstrated that the presence of resident corneal macrophages is essential for production of IL-1beta and CXCL1/KC, and for recruitment of neutrophils and mononuclear cells into the corneal stroma. We found that beta-glucan was highly expressed on germinating conidia and hyphae in the cornea stroma, and that both Dectin-1 and phospho-Syk were up-regulated in infected corneas. Additionally, we show that infected Dectin-1(-/-) corneas have impaired IL-1beta and CXCL1/KC production, resulting in diminished cellular infiltration and fungal clearance compared with control mice, especially during infection with clinical isolates expressing high beta-glucan. In contrast to Dectin 1(-/-) mice, cellular infiltration into infected TLR2(-/-), TLR4(-/-), and MD-2(-/-) mice corneas was unimpaired, indicating no role for these receptors in cell recruitment; however, fungal killing was significantly reduced in TLR4(-/-) mice, but not TLR2(-/-) or MD-2(-/-) mice. We also found that TRIF(-/-) and TIRAP(-/-) mice exhibited no fungal-killing defects, but that MyD88(-/-) and IL-1R1(-/-) mice were unable to regulate fungal growth. In conclusion, these data are consistent with a model in which beta-glucan on A.fumigatus germinating conidia activates Dectin-1 on corneal macrophages to produce IL-1beta, and CXCL1, which together with IL-1R1/MyD88-dependent activation, results in recruitment of neutrophils to the corneal stroma and TLR4-dependent fungal killing.
authors.affiliation: Department of Ophthalmology and Visual Sciences, Case Western Reserve University, Cleveland, Ohio, United States of America.
authors.forename: Sixto M
authors.initials: SM
authors.lastname: Leal
authors.forename: Susan
authors.initials: S
authors.lastname: Cowden
authors.forename: Yen-Cheng
authors.initials: YC
authors.lastname: Hsia
authors.forename: Mahmoud A
authors.initials: MA
authors.lastname: Ghannoum
authors.forename: Michelle
authors.initials: M
authors.lastname: Momany
authors.forename: Eric
authors.initials: E
authors.lastname: Pearlman
mesh_terms: D000818:Animals; D001232:Aspergillus fumigatus / Q000254:growth & development / Q000472:pathogenicity*; D054360:Chemokine CXCL1 / Q000096:biosynthesis; D003319:Corneal Stroma / Q000382:microbiology / Q000473:pathology; D053583:Interleukin-1beta / Q000096:biosynthesis; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism; D007634:Keratitis / Q000382:microbiology*; D037181:Lectins, C-Type; D008264:Macrophages / Q000502:physiology; D008565:Membrane Proteins / Q000502:physiology*; D051379:Mice; D009419:Nerve Tissue Proteins / Q000502:physiology*; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D000072377:Syk Kinase; D051197:Toll-Like Receptor 4 / Q000502:physiology*; D047071:beta-Glucans / Q000378:metabolism; D000099181:Dectin-1
chemical_list: D054360:Chemokine CXCL1; D053583:Interleukin-1beta; D047908:Intracellular Signaling Peptides and Proteins; D037181:Lectins, C-Type; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D011505:Protein-Tyrosine Kinases; D000072377:Syk Kinase; D051197:Toll-Like Receptor 4; D047071:beta-Glucans; D000099181:Dectin-1; C475895:Cxcl1 protein, mouse; C000606897:Syk protein, mouse