Journal of Leukocyte Biology
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Originally published online as doi:10.1189/jlb.0807523 on March 3, 2008

Published online before print March 3, 2008
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(Journal of Leukocyte Biology. 2008;83:1396-1403.)
© 2008 by Society for Leukocyte Biology

Identification and functional characterization of a bovine orthologue to DC-SIGN

Yoshika Yamakawa*, Christopher Pennelegion*, Samuel Willcocks*, Amanda Stalker*, Niall MacHugh{dagger}, Dave Burt{ddagger}, Tracey J. Coffey§ and Dirk Werling*,1

* Department of Pathology and Infectious Diseases, Royal Veterinary College, Hatfield, United Kingdom;
§ Institute for Animal Health, Compton, Berkshire, United Kingdom;
{ddagger} Roslin Institute, Midlothian, United Kingdom; and
{dagger} Centre for Tropical Veterinary Medicine, The Royal (Dick) School of Veterinary Medicine, Easter Bush Veterinary Centre, Roslin, United Kingdom

1Correspondence: Department of Pathology and Infectious Diseases, Royal Veterinary College, Hawkshead Lane, Hatfield, AL9 7TA, UK. E-mail: dwerling{at}rvc.ac.uk

Dendritic cell-specific ICAM-3-grabbing nonintegrin (DC-SIGN) C-type lectin is almost exclusively expressed at the cell surface of DC. In addition to its normal function facilitating contact of DC with T cells, DC-SIGN has been shown to bind a variety of pathogens, including Mycobacterium bovis, and HIV-1 envelope protein gp120. In this study, we identified the bovine ortholog of the human DC-SIGN gene within the bovine genome, which exists as a single copy. PCR amplified a product, showing a 100% match with the predicted sequences as well as a sequence predicted to be similar to that of SIGNR7. Furthermore, a protein with the same molecular weight as human DC-SIGN was detected by Western blot in cell lysate derived from bovine DC. To characterize this molecule functionally, the uptake of FITC-labeled OVA and FITC-labeled gp120 (FITC-gp120) by bovine and human DC was assessed. FITC-gp120 was shown to bind to bovine DC in a time- and temperature-dependent manner. Binding was blocked by a polyclonal anti-DC-SIGN antibody but not by a control antibody. Furthermore, blocking of this molecule also reduced the binding of M. bovis bacillus Calmette-Guerin expressing GFP. Confocal microscopy showed that DC-SIGN was expressed on the surface of bovine DC. Subsequent pulse-chase studies revealed that FITC-gp120 was internalized by bovine monocyte-derived DC as early as 10 min. Thus, there is evidence of a DC-SIGN-like molecule expressed specifically by bovine DC. This molecule may play an important role in the infection of bovine (DC) cells with M. bovis.

Key Words: dendritic cells • C-type lectin • gp120 • Mycobacterium bovis BCG






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