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J. Biol. Chem., Vol. 275, Issue 26, 20157-20167, June 30, 2000
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From the Department of Dermatology, University of Texas
Southwestern Medical Center, Dallas, Texas 75235-9069
Dendritic cells (DC) are special subsets of
antigen presenting cells characterized by their potent capacity to
activate immunologically naive T cells. By subtracting the mRNAs
expressed by the mouse epidermus-derived DC line XS52 with the
mRNAs expressed by the J774 macrophage line, we identified five
novel genes that were expressed selectively by this DC line. One of
these genes encoded a type II membrane-integrated polypeptide of 244 amino acids containing a putative carbohydrate recognition domain motif
at the COOH-terminal end. This molecule, termed "dectin-1," was
expressed abundantly at both mRNA and protein levels by the XS52 DC
line, but not by non-DC lines (including the J774 macrophage line).
Dectin-1 mRNA was detected predominantly in spleen and thymus (by
Northern blotting) and in skin-resident DC, i.e. Langerhans
cells (by reverse transcription-polymerase chain reaction).
Affinity-purified antibody against dectin-1 identified a 43-kDa
glycoprotein in membrane fractions isolated from the XS52 DC line and
from the dectin-1 cDNA-transfected COS-1 cells. His-tagged recombinant
proteins containing the extracellular domains of dectin-1 showed marked
and specific binding to the surface of T cells and promoted their
proliferation in the presence of anti-CD3 monoclonal antibody at
suboptimal concentrations. These in vitro results suggest
that dectin-1 on DC may bind to as yet undefined ligand(s) on T cells,
thereby delivering T cell co-stimulatory signals. Not only do these
results document the efficacy of subtractive cDNA cloning for the
identification of unique genes expressed by DC, they also provide a
framework for studying the physiological function of dectin-1.
The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EMBL Data Bank with accession number(s) AF262985.
Identification of a Novel, Dendritic Cell-associated
Molecule, Dectin-1, by Subtractive cDNA Cloning*
*
This work was supported by National Institutes of Health
Grants RO1-AR44189, RO1-AR35068, RO1-AR43777, and RO1-AI43262; a grant
from Taisho Pharmaceutical Co., Ltd., Ohmiya, Japan; and an award from
the Centre de Recherches et Investigations Epidermiques et Sensorielles
(to A. T.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: Dept. of Dermatology,
University of Texas Southwestern Medical Center, 5323 Harry Hines
Blvd., Dallas, TX 75235-9069. Tel.: 214-648-3419; Fax: 214-648-3472; E-mail: atakas@mednet.swmed.edu.
Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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