title: Phenotypic heterogeneity in the Gray platelet syndrome extends to the expression of TREM family member, TLT-1.
abstract: The Gray platelet syndrome (GPS) is a rare inherited disorder linked to undefined molecular abnormalities that prevent the formation and maturation of alpha-granules. Here, we report studies on two patients from unrelated families that confirm phenotypic heterogeneity in the disease. First we used immunoelectron microscopy (I-EM) to confirm that TREM-like transcript-1 (TLT-1) is mostly localized to alpha-granule membranes of normal platelets. Then we performed Western blotting (WB) and flow cytometry with permeabilized platelets to show that TLT-1 is selectively reduced in the platelets of patient 1, previously noted to be deficient in glycoprotein (GP)VI (Nurden et al., Blood 2004; 104: 107-114). Yet both TLT-1 and GPVI were normally expressed in platelets of patient 2. Usual levels of JAM-C and claudin-5, also members of the immunoglobulin receptor family, were detected in platelets of both patients. In contrast, P-selectin was markedly decreased for patient 1 but not patient 2. Two metalloproteases, MMP-2 and MMP-9 were normally present. As predicted, platelets of patient 1 showed little labelling for TLT-1 in I-EM, whereas residual Fg was seen in small vesicular structures and P-selectin lining vacuoles or channels of what may be elements of the surface-connected canalicular system. Our results identify TLT-1 as a glycoprotein potentially targeted in platelets of GPS patients, while decreases in at least three membrane glycoproteins suggest that an unidentified proteolytic activity may contribute to the phenotype in some patients with this rare disease.
authors.affiliation: Centre de Référence des Pathologies Plaquettaires, Plateforme Technologique et d'Innovation Biomédicale, Hôpital Xavier Arnozan, 33600 Pessac, France. Alan.Nurden@cnrshl.u-bordeaux2.fr
authors.forename: Alan T
authors.initials: AT
authors.lastname: Nurden
authors.forename: Paquita
authors.initials: P
authors.lastname: Nurden
authors.forename: Emilsé
authors.initials: E
authors.lastname: Bermejo
authors.forename: Robert
authors.initials: R
authors.lastname: Combrié
authors.forename: Daniel W
authors.initials: DW
authors.lastname: McVicar
authors.forename: A Valance
authors.initials: AV
authors.lastname: Washington
mesh_terms: D001792:Blood Platelets / Q000378:metabolism* / Q000648:ultrastructure; D015153:Blotting, Western; D018955:CD36 Antigens / Q000097:blood; D015815:Cell Adhesion Molecules / Q000097:blood; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D020778:Matrix Metalloproteinase 2 / Q000097:blood; D020780:Matrix Metalloproteinase 9 / Q000097:blood; D016253:Microscopy, Immunoelectron; D019007:P-Selectin / Q000097:blood; D010641:Phenotype; D010981:Platelet Storage Pool Deficiency / Q000378:metabolism* / Q000473:pathology; D011971:Receptors, Immunologic / Q000097:blood*; D013577:Syndrome
chemical_list: D018955:CD36 Antigens; D015815:Cell Adhesion Molecules; C448000:JAM3 protein, human; D019007:P-Selectin; D011971:Receptors, Immunologic; C473366:TREML1 protein, human; C522361:MMP2 protein, human; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9