title: Effect of platelet calpain on normal T-lymphocyte CD43: hypothesis of events in the Wiskott-Aldrich syndrome.
abstract: The Wiskott-Aldrich syndrome (WAS) is an inherited disease involving defects of platelets (small size, severe thrombocytopenia due to accelerated destruction) and T lymphocytes (progressive immunodeficiency, lymphopenia). The best-characterized molecular defect is the deficiency and, in some cases, abnormal forms of the T-lymphocyte surface mucin molecule CD43; deficiency of the platelet surface mucin GPIb was observed previously in two of four patients. Neither of these defects is primary, since CD43 and GPIb are encoded by autosomal genes and the disease is X-linked. This study uses cellular biological approaches to explore the possibility that destruction of structurally defective WAS platelets, mimicked experimentally by sonication of normal platelets, plays a role by releasing protease and generating other cellular defects. We show that a protease of normal platelets, identified as Ca(2+)-dependent neutral protease (calpain), which is known to cleave platelet GPIb, also specifically cleaves CD43 on the surface of neighboring desialylated T lymphocytes. The identification of the CD43 cleaving protease was based on its requirement for Ca2+ and inhibition by leupeptin, but not by diisopropylfluorophosphate (DFP). The approximate site of CD43 cleavage was identified by the use of a rabbit antibody. Sensitivity of GPIb to calpain is shown to be sialylation-independent and that of CD43 to be sialylation-dependent, and these findings are explained in terms of molecular structures. These and previous findings are incorporated into a putative mechanism, which explains most of the defects in the WAS. The mechanism suggests that the primary defective molecule in the WAS is unlikely to be a surface glycoprotein, but rather a cytoplasmic molecule with a function in cytoskeletal interactions and/or calcium ion regulation and calpain activation.
authors.affiliation: Center for Blood Research, Boston, MA 02115.
authors.forename: E
authors.initials: E
authors.lastname: Remold-O'Donnell
authors.forename: J
authors.initials: J
authors.lastname: Van Brocklyn
authors.forename: D M
authors.initials: DM
authors.lastname: Kenney
mesh_terms: D015703:Antigens, CD; D001792:Blood Platelets / Q000201:enzymology*; D002118:Calcium / Q000494:pharmacology; D002154:Calpain / Q000037:antagonists & inhibitors / Q000378:metabolism*; D006801:Humans; D015151:Immunoblotting; D051917:Leukosialin; D007976:Leupeptins / Q000494:pharmacology; D008954:Models, Biological; D019158:N-Acetylneuraminic Acid; D009439:Neuraminidase / Q000494:pharmacology; D010980:Platelet Membrane Glycoproteins / Q000378:metabolism; D012794:Sialic Acids / Q000097:blood; D012795:Sialoglycoproteins / Q000378:metabolism*; D013379:Substrate Specificity; D013601:T-Lymphocytes / Q000276:immunology*
chemical_list: D015703:Antigens, CD; D051917:Leukosialin; D007976:Leupeptins; D010980:Platelet Membrane Glycoproteins; C465014:SPN protein, human; D012794:Sialic Acids; D012795:Sialoglycoproteins; D009439:Neuraminidase; D002154:Calpain; D019158:N-Acetylneuraminic Acid; C032854:leupeptin; D002118:Calcium