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Ly49B Is Expressed on Multiple Subpopulations of Myeloid Cells¹

Frances Gays^*, Jonathan G. Aust^*, Delyth M. Reid, Jane Falconer^*, Noriko Toyama-Sorimachi, Philip R. Taylor and Colin G. Brooks^2,*

^* Institute of Cell and Molecular Biosciences, The Medical School, Newcastle, United Kingdom; Department of Ophthalmology, University of Aberdeen, United Kingdom; Department of Gastroenterology Research Institute, International Medical Center of Japan, Tokyo, Japan; and Sir William Dunn School of Pathology, Oxford University, Oxford, United Kingdom

Using a novel mAb specific for mouse Ly49B, we report here that Ly49B, the last remaining member of the C57 Ly49 family to be characterized, is expressed at low levels on 1.5% of spleen cells, none which are NK cells or T cells but which instead belong to several distinct subpopulations of myeloid cells defined by expression of CD11b and different levels of Gr1. Much larger proportions of bone marrow and peritoneal cells expressed Ly49B, all being CD11b⁺ and comprising multiple subpopulations defined by light scatter, F4/80, and Gr1 expression. Costaining for Ly49Q, also expressed on myeloid cells, revealed that Ly49B and Ly49Q were most strongly expressed on nonoverlapping subpopulations, Ly49Q^high cells being mostly B220⁺CD4⁺ and/or CD8⁺, Ly49B⁺ cells lacking these markers. Myeloid populations that developed from bone marrow progenitors in vitro frequently coexpressed both Ly49B and Ly49Q, and Ly49B expression could be up-regulated by LPS, -IFN, and -IFN, often independently of Ly49Q. PCR analysis revealed that cultured NK cells and T cells contained Ly49B transcripts, and Ly49B expression could be detected on NK cells cultured in IL-12 plus IL-18, and on an immature NK cell line. Immunohistochemical studies showed that Ly49B expression in tissues overlapped with but was distinct from that of all other myeloid molecules examined, being particularly prominent in the lamina propria and dome of Peyer’s patches, implicating an important role of Ly49B in gut immunobiology. In transfected cells, Ly49B was found to associate with SHP-1, SHP-2, and SHIP in a manner strongly regulated by intracellular phosphorylation events.

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.

¹ This work was generously supported by grants from the Biotechnology and Biological Sciences Research Council, U.K.

² Address correspondence and reprint requests to Dr. Colin G. Brooks, Institute of Cell and Molecular Biosciences, The Medical School, Newcastle NE2 4HH, U.K.

³ Abbreviations used in this paper: BM, bone marrow; cII, MHC class II; CRD, carbohydrate recognition domain; DC, dendritic cell; FL, flt3 ligand; HA, hemagglutinin; SHP, Src homology region 2 domain-containing phosphatase; SSC, side light scatter.

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