title: Integrin alpha 4 beta 1 and glycoprotein IV (CD36) are expressed on circulating reticulocytes in sickle cell anemia.
abstract: The abnormal adherence of red blood cells, especially circulating reticulocytes (erythrocyte precursors), to the endothelium is believed to contribute to vascular occlusion observed in patients with sickle cell disease. Although several plasma proteins including von Willebrand factor and fibronectin have been proposed to mediate this adhesion, the mechanism of sickle cell adhesion to the endothelium remains unknown. Using flow cytometry, we screened sickle red blood cells with monoclonal antibodies (MoAbs) against known adhesion receptors and detected integrin subunits alpha 4 and beta 1 and the nonintegrin glycoprotein IV on reticulocytes but not on erythrocytes. No reactivity was detected against integrin subunits alpha 2, alpha 3, alpha 5, alpha 6, alpha v, beta 2, beta 3, integrin alpha IIb beta 3, or the nonintegrin glycoprotein Ib. Immunoprecipitation of reticulocytes with either alpha 4- or beta 1-specific antibodies identified the alpha 4 beta 1 complex (alpha 4(70) and alpha 4(80) forms), a receptor for fibronectin and vascular cell adhesion molecule-1. An antibody against glycoprotein IV, a receptor reported to bind thrombospondin and collagen, immunoprecipitated an 88-kD protein consistent with its reported M(r). MoAbs against alpha 4 and glycoprotein IV bound to an average of 4,600 and 17,500 sites per reticulocyte, respectively. Identification of alpha 4 beta 1 and glycoprotein IV on reticulocytes suggests both plasma-dependent and independent mechanisms of reticulocyte adhesion to endothelium and exposed extracellular matrix.
authors.affiliation: Department of Pharmacology, University of North Carolina at Chapel Hill 27599-7365.
authors.forename: C C
authors.initials: CC
authors.lastname: Joneckis
authors.forename: R L
authors.initials: RL
authors.lastname: Ackley
authors.forename: E P
authors.initials: EP
authors.lastname: Orringer
authors.forename: E A
authors.initials: EA
authors.lastname: Wayner
authors.forename: L V
authors.initials: LV
authors.lastname: Parise
mesh_terms: D000755:Anemia, Sickle Cell / Q000097:blood*; D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex / Q000302:isolation & purification; D015703:Antigens, CD / Q000096:biosynthesis* / Q000097:blood / Q000302:isolation & purification; D018955:CD36 Antigens; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D004912:Erythrocytes / Q000378:metabolism; D005434:Flow Cytometry; D006801:Humans; D066298:In Vitro Techniques; D039041:Integrin alpha4beta1; D016023:Integrins / Q000096:biosynthesis* / Q000302:isolation & purification; D008970:Molecular Weight; D010980:Platelet Membrane Glycoproteins / Q000096:biosynthesis; D012156:Reticulocytes / Q000378:metabolism*
chemical_list: D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D015703:Antigens, CD; D018955:CD36 Antigens; D039041:Integrin alpha4beta1; D016023:Integrins; D010980:Platelet Membrane Glycoproteins