title: Excretions/secretions from medicinal larvae (Lucilia sericata) inhibit complement activation by two mechanisms.
abstract: Larvae of the blowfly Lucilia sericata facilitate wound healing by removing dead tissue and biofilms from non-healing and necrotic wounds. Another beneficial action of larvae and their excretions/secretions (ES) is down-regulation of excessive inflammation. As prolonged complement activation is key to excessive inflammation, the aim of this study was to elucidate the mechanisms underlying the anti-complement activities of ES. Results revealed that heat sensitive serine proteases in ES degrade multiple complement proteins in all steps of the three complement activation pathways. Importantly, C3a and C5a-major activators of inflammation-were also degraded by ES and pretreatment of these factors with ES completely blocked their ability to induce activation of human neutrophils. Pre-exposure of the neutrophils to ES did not affect their responsiveness to C3a/C5a and fMLP, indicating that the receptors for these activators on neutrophils were not affected by ES. Surprisingly, heat and serine protease inhibitor pretreatment did not affect the ability of ES to inhibit C5b-9 complex formation despite degrading complement proteins, indicating a second complement-inhibiting molecule in ES. Heated ES was as effective as intact ES in inhibiting C3 deposition upon activation of the alternative pathway, but was significantly less effective in wells with a classical or lectin pathway-specific coating. Unfortunately, the molecules affecting the complement system could not be identified due to an insufficient database for L. sericata. Together, larval ES inhibit complement activation by two different mechanisms and down-regulate the C3a/C5a-mediated neutrophil activation. This attenuates the inflammatory process, which may facilitate wound healing.
authors.affiliation: Department of Infectious Diseases, Leiden University Medical Center, Leiden, the Netherlands.|Department of Surgery and Oncology, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan.
authors.forename: Tetsuro
authors.initials: T
authors.lastname: Tamura
authors.affiliation: Department of Surgery, Medical Center Haaglanden/Bronovo Hospital, The Hague, the Netherlands.
authors.forename: Gwendolyn
authors.initials: G
authors.lastname: Cazander
authors.affiliation: Department of Medical Microbiology, University Medical Center Utrecht, Utrecht, the Netherlands.
authors.forename: Suzan H M
authors.initials: SH
authors.lastname: Rooijakkers
authors.affiliation: Department of Rheumatology, Leiden University Medical Center, Leiden, the Netherlands.
authors.forename: Leendert A
authors.initials: LA
authors.lastname: Trouw
authors.affiliation: Department of Infectious Diseases, Leiden University Medical Center, Leiden, the Netherlands.
authors.forename: Peter H
authors.initials: PH
authors.lastname: Nibbering
mesh_terms: D000818:Animals; D012634:Bodily Secretions / Q000378:metabolism; D003167:Complement Activation / Q000502:physiology; D015926:Complement C3a / Q000037:antagonists & inhibitors* / Q000378:metabolism; D015936:Complement C5a / Q000037:antagonists & inhibitors* / Q000378:metabolism; D003646:Debridement / Q000379:methods; D004175:Diptera / Q000378:metabolism*; D015536:Down-Regulation; D007249:Inflammation / Q000378:metabolism / Q000503:physiopathology*; D007814:Larva / Q000502:physiology*; D018375:Neutrophil Activation / Q000502:physiology; D014945:Wound Healing / Q000502:physiology*
chemical_list: D015926:Complement C3a; D015936:Complement C5a