Macrophage MHC receptor 2: a novel receptor on allograft (H-2D(d)K(d))-induced macrophage (H-2D(b)K(b)) recognizing an MHC class I molecule, H-2K(d), in mice.

We previously reported that a population of allograft (H-2D(d)K(d))-induced macrophages (AIM) in C57BL/6 (H-2D(b)K(b)) mice exhibited major histocompatibility complex (MHC) haplotype (H-2D(d)) specific killing of the allograft (e.g., BALB/c skin and Meth A cells; H-2D(d)K(d)) in a macrophage MHC receptor (MMR)-dependent manner. In the present study, we isolated a cDNA clone encoding a novel receptor (MMR2) on AIM recognizing another MHC class I molecule, H-2K(d), by the expression cloning method using H-2K(d) tetramer and a monoclonal antibody (mAb; R12) specific for AIM. The cDNA (2359-bp) encoded a 677-amino acid polypeptide of a calculated molecular mass of 87 kDa and was found to be expressed exclusively on AIM among cells infiltrating into allografts on days 0-9 after transplantation. Confocal microscopy showed that HEK293T cells transfected with this cDNA were reactive toward the H-2K(d) molecule but not toward other MHC class I molecules such as H-2D(d), H-2D(b), H-2D(k), H-2K(b), H-2K(k), and H-2L(d) molecules. The binding of the H-2K(d) molecule to the transfectants was inhibited by the addition of R12 or anti-H-2K(d), but not by R15 (a mAb specific for H-2D(d) receptor) or anti-H-2D(d), mAb. Flow cytometric analysis revealed specific binding of H-2K(d) molecules to AIM (K(d)=2.7x10(-9) M); and the binding was completely suppressed by the addition of R12 mAb. These results demonstrate that a novel receptor (MMR2) for H-2K(d) molecules was induced on effector macrophages responsible for allograft (H-2D(d)K(d)) rejection by H-2D(b)K(b) mice.

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