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* Department of Microbiology and Immunology, Department of Medicine, and Walther Oncology Center, Indiana University School of Medicine, Indianapolis, Indiana;
Laboratory of Bacterial Pathogenesis and Immunology, and
|| Laboratory of Molecular Genetics and Immunology, The Rockefeller University, New York, New York;
Department of Molecular Genetics, Institute for Liver Research, Kansai Medical University, Moriguchi, Japan; and
Institute of Immunobiology, University of Freiburg, Freiburg, Germany
Correspondence: Tie Chen, Department of Microbiology, Immunology and Medicine, Walther Oncology Center, Indiana University School of Medicine, MS 252, 635 Barnhill Dr., Indianapolis, IN 46202-5120. E-mail: tiechen{at}iupui.edu
Biliary glycoprotein (BGP, CD66a, CEACAM1) is a member of the
carcinoembryonic antigen family (CEA, CD66), a group of transmembrane
proteins belonging to the immunoglobulin superfamily. The structural
features surrounding the tyrosine residues in the cytoplasmic domain of
BGP share similarity with the consensus sequence of the immunoreceptor
tyrosine-based inhibition motif (ITIM), the docking site for SHIP,
SHP-1, and SHP-2 molecules. Using the well-characterized inhibitory
receptor, Fc
RIIB, we constructed a FcRIIB-BGPa chimeric molecule
that contained the extracellular and transmembrane domain of FcRIIB
and the cytoplasmic tail of BGPa and expressed it in DT40 B cells. Our
results showed that FcRIIB-BGPa, just like the unmodified FcRIIB
molecule, inhibited calcium influx in activated DT40 B cells.
Substitution of tyrosine with phenylalanine (Y459F) in FcRIIB-BGPa
completely abrogated its ability to inhibit calcium influx, indicating
that the motif surrounding Y459 is ITIM. The presence of ITIM was also
supported by showing that the FcRIIB-BGPa-mediated inhibitory effect
was reduced in SHP-1and SHP-2 mutant DT40 B cells and further
diminished in a SHP-1/-2 double-deficient mutant line. The results
suggest that SHP-1 and SHP-2 are required for the
FcRIIB-BGPa-mediated inhibitory signals.
Key Words: B-cell antigen receptor • FcRs • ITIM • SHIP
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