CD226 mediates platelet and megakaryocytic cell adhesion to vascular endothelial cells.
Platelet adhesion to vascular endothelial cells is a pathophysiologically relevant cell-to-cell interaction. However, the mechanisms underlying this cellular interaction are incompletely understood. In search of the ligand for CD226 adhesion molecule expressed on platelets, we found that human umbilical vein endothelial cells (HUVEC) express significant amount of putative CD226 ligand. We demonstrated that thrombin-activated, but not resting, platelets bind to intact HUVEC. Anti-CD226 monoclonal antibody specifically inhibited the binding, indicating that CD226 mediates the intercellular binding between thrombin-activated platelets and HUVEC. We also demonstrated that platelet activation with thrombin induces tyrosine phosphorylation of CD226 as well as CD226-mediated platelet adhesion. Moreover, experiments using mutant transfectants suggested that the tyrosine at residue 322 of CD226 plays an important role for its adhesive function. CD226 was also expressed on primary megakaryocytes and megakaryocytic cell lines. Anti-CD226 monoclonal antibody inhibited binding of megakaryocytic cell lines to HUVEC. Taken together, these results reveal a novel mechanism for adhesion of platelets and megakaryocytic cells to vascular endothelial cells.
Kojima
Hiroshi
H
Division of Hematology, Institute of Clinical Medicine and Department of Immunology, Institute of Basic Medical Sciences, University of Tsukuba, Tsukuba, Ibaraki 305-8575, Japan. hkojima@md.tsukuba.ac.jp
Kanada
Hirotaka
H
Shimizu
Seiichi
S
Kasama
Emi
E
Shibuya
Kazuko
K
Nakauchi
Hiromitsu
H
Nagasawa
Toshiro
T
Shibuya
Akira
A
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000378:metabolism; D000945:Antigens, Differentiation, T-Lymphocyte / Q000096:biosynthesis* / Q000737:chemistry*; D001792:Blood Platelets / Q000378:metabolism*; D019556:COS Cells; D002448:Cell Adhesion; D002460:Cell Line; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology*; D005434:Flow Cytometry; D005452:Fluoresceins / Q000378:metabolism; D006801:Humans; D008024:Ligands; D008533:Megakaryocytes / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D009154:Mutation; D010766:Phosphorylation; D013917:Thrombin / Q000737:chemistry; D014162:Transfection; D014443:Tyrosine / Q000737:chemistry / Q000378:metabolism; D014471:Umbilical Veins / Q000166:cytology; D000098862:T Lineage-Specific Activation Antigen 1
D000911:Antibodies, Monoclonal; D000945:Antigens, Differentiation, T-Lymphocyte; D000098862:T Lineage-Specific Activation Antigen 1; D005452:Fluoresceins; D008024:Ligands; D014443:Tyrosine; D013917:Thrombin; C007740:fluorexon