title: Molecular characterization and expression of porcine Siglec-5.
abstract: In this study we describe the characterization of the porcine orthologue of Siglec-5. A cDNa clone was obtained from a porcine cDNa library derived from swine small intestine which encodes a 555 a-a type 1 transmembrane protein with sequence homology to human Siglec-5. This protein consists of four Ig-like domains, a transmembrane region, and a cytoplasmic tail with two tyrosine-based signalling motifs. When expressed as a recombinant protein fused to the Fc region of human IgG1, porcine Siglec-5 was able to bind porcine red blood cells in a sialic acid-dependent manner. Monoclonal antibodies (mAb) were developed against porcine Siglec-5 and used to analyse its expression in bone marrow and blood cells, and lymphoid tissues. Porcine Siglec-5 expression was mainly restricted to myelomonocytic cells and their precursors, being detected also, although at low levels, on plasmacytoid dendritic cells and B lymphocytes. In lymphoid tissues, ellipsoids of the spleen and subcapsular and medullar sinuses of lymph nodes were positive for Siglec-5. These mAbs were able to precipitate, from granulocyte lysates, a protein of approximately 85 kDa under non-reducing conditions, indicating that porcine Siglec-5 is expressed as a monomer in the plasma membrane.
authors.affiliation: Dpto. de Biotecnología, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Ctra de la Coruña Km 7.5, 28040 Madrid, Spain.
authors.forename: Z
authors.initials: Z
authors.lastname: Escalona
authors.affiliation: Dpto. de Biotecnología, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Ctra de la Coruña Km 7.5, 28040 Madrid, Spain.
authors.forename: B
authors.initials: B
authors.lastname: Álvarez
authors.affiliation: National Institute of Agrobiological Sciences (NIAS), 2 Ikenodai, Tsukuba, Ibaraki 305-8602, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Uenishi
authors.affiliation: National Institute of Agrobiological Sciences (NIAS), 2 Ikenodai, Tsukuba, Ibaraki 305-8602, Japan.
authors.forename: D
authors.initials: D
authors.lastname: Toki
authors.affiliation: Dpto. de Biotecnología, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Ctra de la Coruña Km 7.5, 28040 Madrid, Spain.
authors.forename: M
authors.initials: M
authors.lastname: Yuste
authors.affiliation: Dpto. de Biotecnología, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Ctra de la Coruña Km 7.5, 28040 Madrid, Spain.
authors.forename: C
authors.initials: C
authors.lastname: Revilla
authors.affiliation: Dpto. de Biología Celular, Facultad de Medicina, Universidad Complutense de Madrid, Avda. Complutense s/n, 28040 Madrid, Spain.
authors.forename: M
authors.initials: M
authors.lastname: Gómez del Moral
authors.affiliation: Dpto. de Biotecnología, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Ctra de la Coruña Km 7.5, 28040 Madrid, Spain.
authors.forename: F
authors.initials: F
authors.lastname: Alonso
authors.affiliation: Dpto. de Biotecnología, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Ctra de la Coruña Km 7.5, 28040 Madrid, Spain.
authors.forename: A
authors.initials: A
authors.lastname: Ezquerra
authors.affiliation: Dpto. de Biotecnología, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Ctra de la Coruña Km 7.5, 28040 Madrid, Spain. Electronic address: juncal@inia.es.
authors.forename: J
authors.initials: J
authors.lastname: Domínguez
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002460:Cell Line; D003001:Cloning, Molecular; D003713:Dendritic Cells / Q000276:immunology*; D004912:Erythrocytes / Q000378:metabolism*; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000235:genetics; D007074:Immunoglobulin G / Q000235:genetics; D008969:Molecular Sequence Data; D022423:Myeloid Cells / Q000276:immunology*; D019158:N-Acetylneuraminic Acid / Q000378:metabolism; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000235:genetics; D063265:Sialic Acid Binding Immunoglobulin-like Lectins / Q000235:genetics / Q000378:metabolism*; D013552:Swine / Q000276:immunology*; D059467:Transcriptome
chemical_list: D000911:Antibodies, Monoclonal; D007141:Immunoglobulin Fc Fragments; D007074:Immunoglobulin G; D011993:Recombinant Fusion Proteins; D063265:Sialic Acid Binding Immunoglobulin-like Lectins; D019158:N-Acetylneuraminic Acid
keywords: Monoclonal antibody; Myeloid cells; Plasmacytoid DCs; Siglec-5; Swine