Recognition of highly restricted regions in the β-propeller domain of αIIb by platelet-associated anti-αIIbβ3 autoantibodies in primary immune thrombocytopenia.
Platelet-associated (PA) IgG autoantibodies play an essential role in primary immune thrombocytopenia (ITP). However, little is known about the epitopes of these Abs. This study aimed to identify critical binding regions for PA anti-αIIbβ3 Abs. Because PA anti-αIIbβ3 Abs bound poorly to mouse αIIbβ3, we created human-mouse chimera constructs. We first examined 76 platelet eluates obtained from patients with primary ITP. Of these, 26 harbored PA anti-αIIbβ3 Abs (34%). Further analysis of 15 patients who provided sufficient materials showed that the epitopes of these Abs were mainly localized in the N-terminal half of the β-propeller domain in αIIb (L1-W235). We could identify 3 main recognition sites in the region; 2 eluates recognized a conformation formed by the W1:1-2 and W2:3-4 loops, 5 recognized W1:2-3, and 4 recognized W3:4-1. The remaining 4 eluates could not be defined by the binding sites. Within these regions, we identified residues critical for binding, including S29 and R32 in W1:1-2; G44 and P45 in W1:2-3; and P135, E136, and R139 in W2:3-4. Of 11 eluates whose recognition sites were identified, 5 clearly showed restricted κ/λ-chain usage. These results suggested that PA anti-αIIbβ3 Abs in primary ITP tended to recognize highly restricted regions of αIIb with clonality.
Department of Hematology and Oncology, Osaka University Graduate School of Medicine, Osaka, Japan.
Kazunobu
K
Kiyomizu
Hirokazu
H
Kashiwagi
Tsuyoshi
T
Nakazawa
Seiji
S
Tadokoro
Shigenori
S
Honda
Yuzuru
Y
Kanakura
Yoshiaki
Y
Tomiyama
D000328:Adult; D000368:Aged; D000595:Amino Acid Sequence; D000818:Animals; D001323:Autoantibodies / Q000276:immunology*; D001665:Binding Sites; D001792:Blood Platelets / Q000276:immunology*; D018360:Crystallography, X-Ray; D000939:Epitopes / Q000276:immunology; D005260:Female; D006801:Humans; D007147:Immunoglobulin Light Chains / Q000378:metabolism; D008297:Male; D051379:Mice; D008875:Middle Aged; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000276:immunology*; D040201:Platelet Membrane Glycoprotein IIb / Q000737:chemistry* / Q000276:immunology*; D011485:Protein Binding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D013921:Thrombocytopenia / Q000276:immunology*
D001323:Autoantibodies; D000939:Epitopes; D007147:Immunoglobulin Light Chains; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D040201:Platelet Membrane Glycoprotein IIb