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Prepublished online as a Blood First Edition Paper on April 17, 2002; DOI 10.1182/blood-2001-12-0265. This Article Full Text Full Text (PDF) All Versions of this Article: 2001-12-0265v1 99/11/4063    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Rights and Permissions Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Walcheck, B. Articles by McEver, R. P. Search for Related Content PubMed PubMed Citation Articles by Walcheck, B. Articles by McEver, R. P. Related Collections Immunobiology Social Bookmarking                   What's this?  Previous Article  |  Table of Contents  |  Next Article  Blood, 1 June 2002, Vol. 99, No. 11, pp. 4063-4069 IMMUNOBIOLOGY The monoclonal antibody CHO-131 binds to a core 2 O-glycan terminated with sialyl-Lewis x, which is a functional glycan ligand for P-selectin Bruce Walcheck, Anne Leppanen, Richard D. Cummings, Randall N. Knibbs, Lloyd M. Stoolman, Shelia R. Alexander, Polly E. Mattila, and Rodger P. McEver From the Department of Veterinary PathoBiology, the Department of Laboratory Medicine and Pathology, and the Center for Immunology, University Minnesota Academic Health Center, University of Minnesota, St Paul; the Department of Biochemistry and Molecular Biology and the Department of Medicine, the Oklahoma Center for Medical Glycobiology; the Warren Medical Research Institute, University of Oklahoma Health Sciences Center; and the Cardiovascular Biology Research Program, Oklahoma Medical Research Foundation, Oklahoma City; and the Department of Pathology, University of Michigan, Ann Arbor. Core 2 O-glycans terminated with sialyl-Lewis x (sLeX) are functionally important oligosaccharides that endow particular macromolecules with high-affinity glycan ligands for the selectin family. To date, antibodies that recognize these structures on leukocytes have not been described. We characterize such a monoclonal antibody (mAb) here (CHO-131). The binding specificity of CHO-131 was directly examined by means of synthetic glycopeptides containing precise O-glycan structures. CHO-131 bound to sLeX extended from a core 2 branch (C2-O-sLeX), but CHO-131 demonstrated no reactivity if this oligosaccharide lacked fucose or if sLeX was extended from a core 1 branch. Using transfected cell lines, we found that CHO-131 binding required the functional activity of the glycosyltransferases 2,3-sialyltransferase, 1,3-fucosyltransferase-VII, and core 2 1,6 N-acetylglucosaminyltransferase (C2GnT). The C2-O-sLeX motif occurs primarily on sialomucins and has been directly shown to contribute to high-affinity P-selectin glycoprotein ligand-1 binding by P-selectin. Indeed, CHO-131 staining of neutrophils was diminished following sialomucin removal by O-glycoprotease, and its reactivity with transfected hematopoietic cell lines correlated with the expression of P-selectin ligands. CHO-131 also stained a small population of lymphocytes that were primarily CD3+, CD4+, and CD45RO+ and represented a subset (37.8% ± 18.3%) of cutaneous lymphocyte-associated antigen (CLA) T cells, distinguished by the mAb HECA-452, which detects sLeX-related glycans. Unlike anti-sLeX mAbs, CHO-131 binding also indicates C2GnT activity and demonstrates that CLA T cells are heterogeneous based on the glycan structures they synthesize. These findings support evidence that differential C2GnT activity results in T-cell subsets that express ligands for E-selectin, P-selectin, or both. © 2002 by The American Society of Hematology.   CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this? This article has been cited by other articles: Y. Wang, A. H. Herrera, Y. Li, K. K. Belani, and B. Walcheck Regulation of Mature ADAM17 by Redox Agents for L-Selectin Shedding J. Immunol., February 15, 2009; 182(4): 2449 - 2457. [Abstract] [Full Text] [PDF] J. J. Miner, L. 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Walcheck ADAM17 deficiency by mature neutrophils has differential effects on L-selectin shedding Blood, October 1, 2006; 108(7): 2275 - 2279. [Abstract] [Full Text] [PDF] P. E. Mattila, C. E. Green, U. Schaff, S. I. Simon, and B. Walcheck Cytoskeletal interactions regulate inducible L-selectin clustering Am J Physiol Cell Physiol, August 1, 2005; 289(2): C323 - C332. [Abstract] [Full Text] [PDF] X. Zou, V. R. Shinde Patil, N. M. Dagia, L. A. Smith, M. J. Wargo, K. A. Interliggi, C. M. Lloyd, D. F. J. Tees, B. Walcheck, M. B. Lawrence, et al. PSGL-1 derived from human neutrophils is a high-efficiency ligand for endothelium-expressed E-selectin under flow Am J Physiol Cell Physiol, August 1, 2005; 289(2): C415 - C424. [Abstract] [Full Text] [PDF] M. J. Smith, B. R. E. Smith, M. B. Lawrence, and K. R. 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