title: Biochemical and immunohistochemical characteristics of CD62 and CD63 monoclonal antibodies. Expression of GMP-140 and LIMP-CD63 (CD63 antigen) in human lymphoid tissues.
abstract: During platelet secretion granule membrane glycoproteins are translocated to the plasma membrane. We report here the biochemical and immunohistochemical characterization of a panel of platelet-secretion-specific, CD62 and CD63 monoclonal antibodies (MoAb), which we raised to thrombin-activated platelets. The CD62 MoAb identify the alpha-granule membrane protein GMP-140, also designated platelet activation-dependent granule external membrane protein (PADGEM). The number of epitopes on thrombin-activated platelets ranged from 15,000 to 20,000. The CD63 MoAb recognize a 30-60 kDalton integral membrane protein of lysosomes. Due to its distinct localization, we have designated the CD63 antigen lysosome integral membrane protein, CD63 (LIMP-CD63). The number of epitopes on thrombin-activated platelets ranged from 9000 to 11,000. Expression of GMP-140, a member of the Selectin family (also referred as the LEC-CAM family) of adhesion molecules, and LIMP-CD63 was examined on human spleen, thymus and lymph node by immunohistochemistry. Both GMP-140 and LIMP-CD63 showed a wide distribution in lymphoid tissues; vascular endothelial cells and tissue compartments that were readily accessible to blood-borne components were uniformly positive for GMP-140 and LIMP-CD63. Furthermore, LIMP-CD63 was expressed in polymorphonuclear granulocytes and macrophages.
authors.affiliation: Department of Haematology, University Hospital Utrecht, The Netherlands.
authors.forename: M J
authors.initials: MJ
authors.lastname: Metzelaar
authors.forename: H J
authors.initials: HJ
authors.lastname: Schuurman
authors.forename: H F
authors.initials: HF
authors.lastname: Heijnen
authors.forename: J J
authors.initials: JJ
authors.lastname: Sixma
authors.forename: H K
authors.initials: HK
authors.lastname: Nieuwenhuis
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry*; D015703:Antigens, CD / Q000032:analysis*; D001792:Blood Platelets / Q000737:chemistry; D006801:Humans; D007150:Immunohistochemistry; D008198:Lymph Nodes / Q000737:chemistry; D008221:Lymphoid Tissue / Q000737:chemistry*; D019007:P-Selectin; D010980:Platelet Membrane Glycoproteins / Q000032:analysis*; D013154:Spleen / Q000737:chemistry; D060149:Tetraspanin 30; D013950:Thymus Gland / Q000737:chemistry
chemical_list: D000911:Antibodies, Monoclonal; D015703:Antigens, CD; C554586:CD63 protein, human; D019007:P-Selectin; D010980:Platelet Membrane Glycoproteins; D060149:Tetraspanin 30