MV-H was synthesized and were colocalized with CD46 in the center of the syncytia regardless of moesin expression (data not shown)
MV-H was synthesized and were colocalized with CD46 in the center of the syncytia regardless of moesin expression (data not shown). (7, BIIB021 10, 31, 33, 34) and moesin (11, 37) have been suggested to be implicated in measles virus (MV) entry. These two molecules are expressed on most human cells, consistent with the wide tissue tropism of MV. CHO cells, otherwise nonpermissive to MV, efficiently form syncytia on transfection with CD46 cDNA (10, 15). Rabbit anti-human CD46 antibody (Ab) and monoclonal Abs (MAbs) against human CD46 recognizing SCR2 block MV-mediated syncytium formation (16, 29, 40). Deglycosylation studies also support the importance of the sugars in SCR2 for MV contamination (28). These results unequivocally indicate that CD46 serves as a receptor for MV. Since CD46 plays a primary role in the protection of host cells from homologous complement (20), it encompasses receptors for the complement system and viral contamination. Evidence supporting the role of moesin as a receptor for MV, on the other hand, seems to remain inconclusive. Moesin is usually a member of the ezrin-radixin-moesin family of proteins, which sustain cell surface molecules and the cytoskeleton (1, 2, 5, 24, 36, 44C46). Moesin is usually widely distributed as an essential intracellular BIIB021 element in cells of various species. It was reported that a MAb against a human astrocytoma cell line (U-251), named 119, inhibited MV contamination and recognized a 75-kDa protein, which was identified as moesin (11). This result was confirmed with other MAbs against moesin and various cell lines of human, monkey, and murine origin (37). Indeed, murine cells with no detectable CD46 homolog were permissive to MV, although far less so than human cells (10, 12, 33, 48), and transfection of human CD46 conferred higher susceptibility to MV (10, 33, 48). These studies indicated that some murine cell lines that can be readily infected with MV must express receptor molecules other than CD46, and moesin is usually a candidate for such an alternative receptor molecule (6, 11, 12, 37). No structural homolog of CD46 has been found in these murine cell lines, and CRRY, a murine functional but not structural homolog of CD46 (14, 19, 25) in terms of complement regulation, is not involved in the entry of MV (12). Further supporting this issue is the fact that murine moesin is usually 98.3% identical to human moesin at the amino acid level (36), reasonably serving as a functional homolog (19, 25), while murine CRRY is 40% homologous to human CD46 (14, 25). However, Ab blocking studies are sometimes difficult to interpret. In fact, Devaux and Gerlier (8) recently suggested that this cross-reactivity of antimoesin Abs with CD46 might explain the inhibitory effects of these Abs on MV entry. If this is the case, moesin, even though it forms a receptor complex with CD46 under the inner leaflet of membranes, may not be directly involved in MV binding. To obtain conclusive evidence, MV infection studies were BIIB021 performed with moesin-positive and -unfavorable embryonic stem (ES) cells expressing or not expressing human CD46. MATERIALS AND METHODS Cells and Abs. CHO cells were obtained from the American Type Culture Collection. Vero cells and MV, a modified Nagahata strain (15, 16), which underwent four passages in hamster brain, were obtained from the Research Institute for Microbial Diseases, Osaka University. Anti-CD46 MAbs M160 and M177 (39) were prepared as described previously. The MAb against MV-H protein was a kind gift from S. Kinesin1 antibody Ueda. A rat anti-mouse moesin MAb, M22.