Molecular and functional characterization of porcine Siglec-3/CD33 and analysis of its expression in blood and tissues
A cDNA clone encoding a 380 a-a type 1 transmembrane protein with homology to human Siglec-3/CD33 was obtained from a swine small intestine library. An analysis of protein sequence identified two immunoglobulin-like domains, a transmembrane region, and a carboxi-terminal tail with two tyrosine-based signalling motifs. Binding assays of Siglec-3 transfected CHO cells to polyacrylamide glycoconjugates showed a preference for α2-6-linked sialic acids. Using mAbs raised against a fragment containing the two Ig-like domains, porcine Siglec-3 was found to be expressed on monocytes and granulocytes, and their bone marrow precursors. It was also detected in lymph node, splenic and alveolar macrophages. MAbs immunoprecipitated, from granulocyte lysates, a protein of 51-60 kDa under both non-reducing and reducing conditions. MAbs were also used to analyse functional activity of Siglec-3 on bone marrow and blood cells. Engagement of Siglec-3 by mAb had no apparent effect on cell proliferation or cytokine production. © 2015 Elsevier Ltd.
Main Authors: | Álvarez, Belén, Escalona, Z., Uenishi, H., Toki, D., Revilla Calvo, Concepción, Yuste, M., Gómez del Moral, M., Alonso, F., Ezquerra Martínez, Ángel, Domínguez, J. |
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Format: | artículo biblioteca |
Language: | English |
Published: |
Elsevier
2015
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Subjects: | Siglec-3, CD33, Monoclonal antibody, Swine, Myeloid cells, Monocyte, |
Online Access: | http://hdl.handle.net/10261/291488 |
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