title: B-CAM/LU expression and the role of B-CAM/LU activation in binding of low- and high-density red cells to laminin in sickle cell disease.
abstract: Red blood cells from patients with sickle cell disease (SS RBC) adhere to laminin and over-express the high-affinity laminin receptor basal cell adhesion molecule/Lutheran protein (B-CAM/LU). This receptor has recently been shown to undergo activation in vitro through a protein kinase A-dependent mechanism. Low-density SS RBC express two-thirds more B-CAM/LU than high-density SS RBC. However, high-density SS RBC have been identified as most adherent to laminin under flow conditions. We investigated the ability of low- and high-density SS RBC to interact with laminin under various conditions and explored factors that might be responsible for the differences in B-CAM/LU-laminin interaction between high- and low-density SS RBC. We confirmed that high-density SS RBC adhere to laminin more strongly than low-density SS RBC under flow conditions. However, low-density SS RBC bind soluble laminin most strongly and are the most adherent to laminin under static conditions. Soluble recombinant Lutheran extracellular domain protein completely blocked SS RBC adhesion to laminin under both static and flow conditions. The protein kinase A inhibitor 14-22 amide inhibited adhesion to laminin during flow by high-density SS RBC from patients with strongly adherent cells but had no effect on adhesion observed after a static phase. Deletion of the cytoplasmic domain of B-CAM as well as mutation of the juxtamembranous tyrosine residue failed to reduce B-CAM-mediated adhesion to laminin by transfected MEL cells. These studies confirm that B-CAM/LU is the most critical receptor mediating adhesion to laminin under both static and flow conditions. Dense SS RBC are most adherent to laminin despite bearing fewer laminin receptors, apparently due to a reversible protein kinase A-dependent process that is unlikely to involve direct phosphorylation of B-CAM/LU. Our results also suggest that the nature of the interaction of B-CAM/LU with laminin may be different under static and flow conditions.
authors.affiliation: Division of Hematology, Department of Medicine, Duke University Medical Center and Duke Comprehensive Sickle Cell Center, Durham, North Carolina 27710, USA.
authors.forename: Qin
authors.initials: Q
authors.lastname: Zen
authors.forename: Milena
authors.initials: M
authors.lastname: Batchvarova
authors.forename: Christina A
authors.initials: CA
authors.lastname: Twyman
authors.forename: Christine E
authors.initials: CE
authors.lastname: Eyler
authors.forename: Huiling
authors.initials: H
authors.lastname: Qiu
authors.forename: Laura M
authors.initials: LM
authors.lastname: De Castro
authors.forename: Marilyn J
authors.initials: MJ
authors.lastname: Telen
mesh_terms: D000328:Adult; D000755:Anemia, Sickle Cell / Q000097:blood*; D001483:Base Sequence; D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000097:blood* / Q000235:genetics*; D017931:DNA Primers; D004912:Erythrocytes / Q000378:metabolism*; D005434:Flow Cytometry; D006801:Humans; D007797:Laminin / Q000097:blood*; D008188:Lutheran Blood-Group System; D009363:Neoplasm Proteins / Q000097:blood* / Q000235:genetics*; D012156:Reticulocytes / Q000378:metabolism / Q000473:pathology
chemical_list: C090228:BCAM protein, human; D015815:Cell Adhesion Molecules; D017931:DNA Primers; D007797:Laminin; D008188:Lutheran Blood-Group System; D009363:Neoplasm Proteins