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Blood, 1 May 2006, Vol. 107, No. 9, pp. 3600-3608.
Prepublished online as a Blood First Edition Paper on January 5, 2006; DOI 10.1182/blood-2005-09-3842.
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Submitted September 27, 2005
Accepted December 15, 2005
Characterisation of Siglec-H as a novel endocytic receptor expressed on murine plasmacytoid dendritic cell precursors
Jiquan Zhang, Anna Raper, Noriko Sugita, Ravi Hingorani, Mariolina Salio, Michael J Palmowski, Vincenzo Cerundolo, and Paul R Crocker*
Division of Cell Biology and Immunology, School of Life Sciences, University of Dundee, Dundee, Scotland, United Kingdom
Division of Periodontology, Department of Oral Biological Science, Niigata University Graduate School of Medical and Dental Sciences, Niigata, Japan
BDBiosciences, San Diego, California, USA
MRC Human Immunology Unit, Weatherall Institute for Molecular Medicine, University of Oxford, Oxford, England, United Kingdom
* Corresponding author; email: p.r.crocker{at}dundee.ac.uk.
We describe the cloning and characterization of Siglec-H, a novel murine CD33-related siglec-like molecule with 2 Ig-domains. Unlike other CD33-related siglecs, Siglec-H lacks tyrosine based signaling motifs in its cytoplasmic tail. Although Siglec-H has the typical structural features required for sialic acid binding, no evidence for carbohydrate recognition was obtained. Specific monoclonal and polyclonal antibodies (Abs) were raised to Siglec-H and used to define its cellular expression pattern and functional properties. By flow cytometry, Siglec H was expressed specifically on plasmacytoid dendritic cell precursors (pDCs) in bone marrow, spleen, blood and lymph node. Staining of tissue sections showed that Siglec-H was also expressed in a subset of marginal zone macrophages in the spleen and in medullary macrophages in lymph nodes. Using bone marrow-derived pDCs that express Siglec-H, addition of Abs did not influence cytokine production, either in the presence or absence of synthetic oligodeoxynucleotides containing unmethylated cytosine-guanine motifs (CpG). In comparison, Siglec-H functioned as an endocytic receptor and mediated efficient internalization of anti-Siglec-H Abs. By immunizing mice with ovalbumin-conjugated anti-Siglec-H Ab in the presence of CpG, we demonstrate generation of antigen-specific CD8 T cells in vivo. Targeting Siglec-H may therefore be a useful way of delivering antigens to pDCs for cross-presentation.
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