title: Genetically engineered fusion of MAP-1 and factor H domains 1-5 generates a potent dual upstream inhibitor of both the lectin and alternative complement pathways.
abstract: Inhibition of the complement cascade has emerged as an option for treatment of a range of diseases. Mannose-binding lectin/ficolin/collectin-associated protein (MAP-1) is a pattern recognition molecule (PRM)-associated inhibitor of the lectin pathway. The central regulator of the alternative pathway (AP) is complement factor H (FH). Our aim was to design a dual upstream inhibitor of both human lectin and APs by fusing MAP-1 with a part of FH. There were 2 different recombinant chimeric proteins comprising full-length human MAP-1 and the first 5 N-terminal domains of human FH designed. The FH domains were orientated either in the N- or C-terminal part of MAP-1. The complement inhibition potential in human serum was assessed. Both chimeric constructs displayed the characteristics of the native molecules and bound to the PRMs with an EC50 of ∼ 2 nM. However, when added to serum diluted 1:4 in a solid-phase functional assay, only the first 5 N-terminal domains of complement FH fused to the C-terminal part of full-length MAP-1 chimeric construct were able to combine inhibition of lectin and AP activation with an half maximal inhibitory concentration of ∼ 100 and 20 nM, respectively. No effect was seen on the classical pathway. Fusion of MAP-1 with FH domains represents a novel therapeutic approach for selective targeting upstream and central complement activation at sites of inflammation.
authors.affiliation: Laboratory of Molecular Medicine, Department of Clinical Immunology-7631, Rigshospitalet, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Mie Anemone
authors.initials: MA
authors.lastname: Nordmaj
authors.affiliation: Laboratory of Molecular Medicine, Department of Clinical Immunology-7631, Rigshospitalet, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Lea
authors.initials: L
authors.lastname: Munthe-Fog
authors.affiliation: Laboratory of Molecular Medicine, Department of Clinical Immunology-7631, Rigshospitalet, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Estrid
authors.initials: E
authors.lastname: Hein
authors.affiliation: Laboratory of Molecular Medicine, Department of Clinical Immunology-7631, Rigshospitalet, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Mikkel-Ole
authors.initials: MO
authors.lastname: Skjoedt
authors.affiliation: Laboratory of Molecular Medicine, Department of Clinical Immunology-7631, Rigshospitalet, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark garred@post5.tele.dk.
authors.forename: Peter
authors.initials: P
authors.lastname: Garred
mesh_terms: D002850:Chromatography, Gel; D017242:Complement Factor H / Q000235:genetics*; D003165:Complement System Proteins / Q000378:metabolism*; D005822:Genetic Vectors; D006801:Humans; D037102:Lectins / Q000378:metabolism*; D037601:Mannose-Binding Lectin / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics*
chemical_list: D037102:Lectins; D037601:Mannose-Binding Lectin; D011993:Recombinant Fusion Proteins; D017242:Complement Factor H; D003165:Complement System Proteins
keywords: chimeric molecules; complement therapeutics; inflammation; recombinant protein; regulation