title: Porcine sialoadhesin (CD169/Siglec-1) is an endocytic receptor that allows targeted delivery of toxins and antigens to macrophages.
abstract: Sialoadhesin is exclusively expressed on specific subpopulations of macrophages. Since sialoadhesin-positive macrophages are involved in inflammatory autoimmune diseases, such as multiple sclerosis, and potentially in the generation of immune responses, targeted delivery of drugs, toxins or antigens via sialoadhesin-specific immunoconjugates may prove a useful therapeutic strategy. Originally, sialoadhesin was characterized as a lymphocyte adhesion molecule, though recently its involvement in internalization of sialic acid carrying pathogens was shown, suggesting that sialoadhesin is an endocytic receptor. In this report, we show that porcine sialoadhesin-specific antibodies and F(ab')₂ fragments trigger sialoadhesin internalization, both in primary porcine macrophages and in cells expressing recombinant porcine sialoadhesin. Using chemical inhibitors, double immunofluorescence stainings and dominant-negative constructs, porcine sialoadhesin internalization was shown to be clathrin- and Eps15-dependent and to result in targeting to early endosomes but not lysosomes. Besides characterizing the sialoadhesin endocytosis mechanism, two sialoadhesin-specific immunoconjugates were evaluated. We observed that porcine sialoadhesin-specific immunotoxins efficiently kill sialoadhesin-expressing macrophages. Furthermore, porcine sialoadhesin-specific albumin immunoconjugates were shown to be internalized in macrophages and immunization with these immunoconjugates resulted in a rapid and robust induction of albumin-specific antibodies, this compared to immunization with albumin alone. Together, these data expand sialoadhesin functionality and show that it can function as an endocytic receptor, a feature that cannot only be misused by sialic acid carrying pathogens, but that may also be used for specific targeting of toxins or antigens to sialoadhesin-expressing macrophages.
authors.affiliation: Laboratory of Virology, Department of Virology, Parasitology, and Immunology, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium. Peter.Delputte@Ugent.be
authors.forename: Peter L
authors.initials: PL
authors.lastname: Delputte
authors.forename: Hanne
authors.initials: H
authors.lastname: Van Gorp
authors.forename: Herman W
authors.initials: HW
authors.lastname: Favoreel
authors.forename: Inge
authors.initials: I
authors.lastname: Hoebeke
authors.forename: Iris
authors.initials: I
authors.lastname: Delrue
authors.forename: Hannah
authors.initials: H
authors.lastname: Dewerchin
authors.forename: Frank
authors.initials: F
authors.lastname: Verdonck
authors.forename: Bruno
authors.initials: B
authors.lastname: Verhasselt
authors.forename: Eric
authors.initials: E
authors.lastname: Cox
authors.forename: Hans J
authors.initials: HJ
authors.lastname: Nauwynck
mesh_terms: D000818:Animals; D000941:Antigens / Q000378:metabolism*; D002352:Carrier Proteins / Q000378:metabolism / Q000502:physiology; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004705:Endocytosis / Q000502:physiology; D000922:Immunotoxins / Q000494:pharmacology; D016676:Macrophages, Alveolar / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000378:metabolism / Q000502:physiology*; D021381:Protein Transport / Q000502:physiology; D011956:Receptors, Cell Surface / Q000378:metabolism; D011971:Receptors, Immunologic / Q000378:metabolism / Q000502:physiology*; D063267:Sialic Acid Binding Ig-like Lectin 1; D013552:Swine; D014118:Toxins, Biological / Q000378:metabolism*; D022161:Transport Vesicles / Q000378:metabolism*
chemical_list: D000941:Antigens; D002352:Carrier Proteins; D000922:Immunotoxins; D008562:Membrane Glycoproteins; D011956:Receptors, Cell Surface; D011971:Receptors, Immunologic; D063267:Sialic Acid Binding Ig-like Lectin 1; D014118:Toxins, Biological