Molecular characterization and expression of porcine Siglec-5 |
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Authors: | Z. Escalona,B. Á lvarez,H. Uenishi,D. Toki,M. Yuste,C. Revilla,M. Gó mez del Moral,F. Alonso,A. Ezquerra,J. Domí nguez |
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Affiliation: | 1. Dpto. de Biotecnología, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Ctra de la Coruña Km 7.5, 28040 Madrid, Spain;2. National Institute of Agrobiological Sciences (NIAS), 2 Ikenodai, Tsukuba, Ibaraki 305-8602, Japan;3. Dpto. de Biología Celular, Facultad de Medicina, Universidad Complutense de Madrid, Avda. Complutense s/n, 28040 Madrid, Spain |
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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. |
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Keywords: | Siglec-5 Monoclonal antibody Swine Myeloid cells Plasmacytoid DCs |
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