Inhibition of NKp30-and 2B4-mediated NK cell activation by evolutionary different human and bovine CEACAM1 receptors

Carcinoembryonicantigen-related cell adhesion molecule 1 (CEACAM1) is a receptor involved in the regulation of NK-cell function. In most species, the CEACAM1 cytoplasmic tail possesses a membrane-proximal ITIM paired with a membrane-distal immunoreceptor tyrosine-based switch motif (ITSM) signaling motif. Human CEACAM1 has phylogenetically relatively recently acquired a second ITIM instead of the ITSM and was shown to inhibit NKG2D-mediated NK-cell activation. Here, we compare the function of bovine and human CEACAM1. We show that in addition to NKG2D, human CEACAM1 can inhibit NK-cell activation via NKp30 or 2B4. Bovine CEACAM1, possessing an ITIM and an ITSM signaling motif, is also inhibitory. However, bovine CEACAM1 inhibition of NKp30-mediated lysis is less pronounced compared with its human counterpart. Bovine CEACAM1 inhibition is dependent on the membrane-proximal ITIM and our data suggest that also the membrane distal ITSM motif contributes to inhibitory signaling. Biochemically, human and bovine CEACAM1 can recruit the phosphatases SHP-1 and SHP-2 after receptor phosphorylation to a similar extend. Bovine CEACAM1 can additionally recruit the adapter molecule Ewing's sarcoma virus-activated transcript-2 (EAT-2), but not SLAM-associated protein (SAP). Taken together, we show that although human and bovine CEACAM1 are differentially equipped with ITIM and ITSM motifs, both receptors can inhibit NKp30 and 2B4 activation of NK cells.