title: Microbial metalloproteinases mediate sensing of invading pathogens and activate innate immune responses in the lepidopteran model host Galleria mellonella.
abstract: Thermolysin-like metalloproteinases such as aureolysin, pseudolysin, and bacillolysin represent virulence factors of diverse bacterial pathogens. Recently, we discovered that injection of thermolysin into larvae of the greater wax moth, Galleria mellonella, mediated strong immune responses. Thermolysin-mediated proteolysis of hemolymph proteins yielded a variety of small-sized (<3 kDa) protein fragments (protfrags) that are potent elicitors of innate immune responses. In this study, we report the activation of a serine proteinase cascade by thermolysin, as described for bacterial lipopolysaccharides (LPS), that results in subsequent prophenoloxidase activation leading to melanization, an elementary immune defense reaction of insects. Quantitative real-time reverse transcription-PCR analyses of the expression of immune-related genes encoding the inducible metalloproteinase inhibitor, gallerimycin, and lysozyme demonstrated increased transcriptional rates after challenge with purified protfrags similar to rates after challenge with LPS. Additionally, we determined the induction of a similar spectrum of immune-responsive proteins that were secreted into the hemolymph by using comparative proteomic analyses of hemolymph proteins from untreated larvae and from larvae that were challenged with either protfrags or LPS. Since G. mellonella was recently established as a valuable pathogenicity model for Cryptococcus neoformans infection, the present results add to our understanding of the mechanisms of immune responses in G. mellonella. The obtained results support the proposed danger model, which suggests that the immune system senses endogenous alarm signals during infection besides recognition of microbial pattern molecules.
authors.affiliation: Institute of Phytopathology and Applied Zoology, Justus-Liebig-University of Giessen, Heinrich-Buff-Ring 26-32, 35392 Giessen, Germany.
authors.forename: Boran
authors.initials: B
authors.lastname: Altincicek
authors.forename: Monica
authors.initials: M
authors.lastname: Linder
authors.forename: Dietmar
authors.initials: D
authors.lastname: Linder
authors.forename: Klaus T
authors.initials: KT
authors.lastname: Preissner
authors.forename: Andreas
authors.initials: A
authors.lastname: Vilcinskas
mesh_terms: D000818:Animals; D015180:Electrophoresis, Gel, Two-Dimensional; D004789:Enzyme Activation; D015870:Gene Expression; D006458:Hemolymph / Q000276:immunology* / Q000378:metabolism; D007113:Immunity, Innate; D007814:Larva; D007915:Lepidoptera / Q000276:immunology* / Q000382:microbiology*; D008070:Lipopolysaccharides / Q000276:immunology; D008666:Metalloendopeptidases / Q000276:immunology / Q000378:metabolism; D018448:Models, Immunological; D009842:Oligopeptides / Q000276:immunology / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012697:Serine Endopeptidases / Q000276:immunology / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013820:Thermolysin / Q000276:immunology* / Q000378:metabolism
chemical_list: D008070:Lipopolysaccharides; D009842:Oligopeptides; D012697:Serine Endopeptidases; D008666:Metalloendopeptidases; C044194:microbial metalloproteinases; D013820:Thermolysin