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Christopher Sassetti^*,, Kirsten Tangemann^*,, Mark S. Singer^*,, David B. Kershaw^,||, and Steven D. Rosen^*,

From the ^* Department of Anatomy and the Program in Immunology, University of California, San Francisco, San Francisco, California 94143; and the Department of Pediatrics and the ^|| Department of Communicable Disease, University of Michigan, Ann Arbor, Michigan 48109

The leukocyte adhesion molecule, L-selectin, mediates the recruitment of lymphocytes to secondary lymphoid organs via interactions with specific ligands presented on high endothelial venules (HEV). Although the HEV-derived ligands for L-selectin are still incompletely defined, they share a common sialomucin-like structure which is thought to present clustered oligosaccharides to the lectin domain of L-selectin. Podocalyxin-like protein (PCLP) is a transmembrane sialomucin that is similar in structure to the well-characterized L-selectin ligand CD34. PCLP has been shown previously to be expressed on the foot processes of podocytes in the kidney glomerulus as well as on vascular endothelium at some sites. We have determined that PCLP is present on HEV, where it binds to both recombinant L-selectin and the HEV-specific monoclonal antibody MECA-79. Furthermore, purified HEV-derived PCLP is able to support the tethering and rolling of lymphocytes under physiological flow conditions in vitro. These results suggest a novel function for PCLP as an adhesion molecule and allow the definition of conserved structural features in PCLP and CD34, which may be important for L-selectin ligand function.

Key Words: podocalyxin • L-selectin • CD34 • high endothelial venule • homing

For their help in obtaining surgical specimens, we would like to thank the members of the Departments of Otolaryngology at the University of California, San Francisco (Drs. Murr, Lee, Lalwani, Dedo, and Schindler); Kaiser Permanente, San Francisco, CA (Dr. Dimeling); and Kaiser Permanente, Marin, CA (Drs. Mizano, Bauman, and Delfanti). In addition, we would especially like to thank the surgical support and scheduling personnel for their invaluable assistance.

Abbreviations used: GlyCAM-1, glycosylation-dependent cell adhesion molecule-1; HEC, high endothelial cell(s); HEV, high endothelial venule(s); HPRT, hypoxanthine phosphoribosyltransferase; HUVEC, human umbilical vein endothelial cell(s); MAdCAM-1, mucosal addressin cell adhesion molecule-1; OSGE, O-sialoglycoprotein endopeptidase; PBS, Dulbecco's PBS; PBST, PBS-Tween; PBS-TX, PBS–Triton X-100; PCLP, podocalyxin-like protein; PNAd, peripheral node addressin.

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