Identification and Characterization of a Novel Human Myeloid Inhibitory C-type Lectin-like Receptor (MICL) That Is Predominantly Expressed on Granulocytes and Monocytes

doi:10.1074/jbc.M313127200

Identification and Characterization of a Novel Human Myeloid Inhibitory C-type Lectin-like Receptor (MICL) That Is Predominantly Expressed on Granulocytes and Monocytes*

^‡Sir William Dunn School of Pathology, University of Oxford, Oxford OX1 3RE, United Kingdom and the ^§Department of Surgery, James H. Quillen College of Medicine, Johnson City, Tennessee 37614

¶ To whom correspondence should be addressed: Sir William Dunn School of Pathology, University of Oxford, South Parks Rd., Oxford OX1 3RE, UK. Tel.: 44-1865-275-531; Fax: 44-1865-275-515; E-mail: gordon.brown{at}path.ox.ac.uk.

Abstract

Inhibitory and activatory C-type lectin-like receptors play an important role in immunity through the regulation of leukocytes. Here, we report the identification and characterization of a novel myeloid inhibitory C-type lectin-like receptor (MICL) whose expression is primarily restricted to granulocytes and monocytes. This receptor, which contains a single C-type lectin-like domain and a cytoplasmic immunoreceptor tyrosine-based inhibitory motif, is related to LOX-1 (lectin-like receptor for oxidized low density lipoprotein-1) and the β-glucan receptor (Dectin-1) and is variably spliced and highly N-glycosylated. We demonstrate that it preferentially associates with the signaling phosphatases SHP-1 and SHP-2, but not with SHIP. Novel chimeric analyses with a construct combining MICL and the β-glucan receptor show that MICL can inhibit cellular activation through its cytoplasmic immunoreceptor tyrosine-based inhibitory motif. These data suggest that MICL is a negative regulator of granulocyte and monocyte function.

Footnotes

↵1 The abbreviations used are: ITAM, immunoreceptor tyrosine-based activation motif; ITIM, immunoreceptor tyrosine-based inhibitory motif; CTLD, C-type lectin-like domain; LOX-1, lectin-like receptor for oxidized low density lipoprotein-1; BGR, β-glucan receptor; MICL, myeloid inhibitory C-type lectin-like receptor; PMN, polymorphonuclear leukocyte; SHP, Src homology-2 domain-containing tyrosine phosphatase; CHO, Chinese hamster ovary; HA, hemagglutinin; RT, reverse transcription; TNF-α, tumor necrosis factor-α; FITC, fluorescein isothiocyanate; PBS, phosphate-buffered saline; SHIP, Src homology-2 domain-containing inositol phosphatase; contig, group of overlapping clones.
↵2 Available at ctld.glycob.ox.ac.uk/.
↵3 J. Herre, A. S. J. Marshall, E. Caron, A. D. Edwards, D. L. Williams, E. Schweighoffer, V. L. Tybulewicz, C. Reis e Sousa, S. Gordon, and G. D. Brown, manuscript in preparation.
↵4 Available at ca.expasy.org/, www.justbio.com/tools.php, and www.ncbi.nlm.nih.gov/BLAST/.
↵5 Available at biochem.boehringer-mannheim.com/prod_inf/manuals/epitope/p20.pdf.
↵6 Available at www.upstate.com/misc/protocols.q.prot.e.activation/Activation+of+Sodium+Orthovanadate.
↵7 Available at www.cbs.dtu.dk/services/NetNGlyc/ and www.cbs.dtu.dk/services/NetOGlyc/.
↵8 E. J. Evans, personal communication.
↵9 Available at bloodsage.gi.k.u-tokyo.ac.jp/.
↵10 Available at www.ncbi.nlm.nih.gov/SAGE.
↵11 E. P. McGreal, G. D. Brown, S. Heinsbroek, S. Zamze, S. Y. Wong, S. Gordon, L. Martinez-Pomares, and P. R. Taylor, manuscript in preparation.
↵* This work was supported by the Theodore Williams Scholarship, the Arthritis Research Council, the British Heart Foundation, the Medical Research Council, and the Wellcome Trust. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank™/EBI Data Bank with accession number(s) AY498550, AY498551, and AY498552.
- Received December 2, 2003.
- Revision received January 9, 2004.
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