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Polymorphism of human platelet membrane glycoprotein IIb associated with
the Baka/Bakb alloantigen system
S Lyman, RH Aster, GP Visentin and PJ Newman
Blood Center of Southeastern Wisconsin, Milwaukee 53233.
The human Baka/Bakb alloantigen system has been implicated in the
pathogenesis of post-transfusion purpura and neonatal alloimmune
thrombocytopenic purpura. Human alloantisera specific for either the Baka
or Bakb allele have been shown to react exclusively with the heavy chain of
membrane glycoprotein (GP) IIb. To investigate the structure of the Bak
epitopes, we used the polymerase chain reaction (PCR) to amplify GPIIb cDNA
synthesized from platelet RNA samples prepared from individuals of known
serologic phenotype. Subsequent DNA sequence analysis of amplified GPIIb
cDNAs derived from one Baka homozygous individual and one Bakb homozygous
individual revealed a single nucleotide base difference near the 3' end of
the mRNA encoding the GPIIb heavy chain. Short 13 base allele-specific
oligonucleotides (ASO) containing the putative phenotype-specific base in
the middle were then synthesized, end-labeled with digoxigenin-11-dUTP
using terminal transferase, and used as probes in subsequent dot-blot
hybridization experiments. Platelet RNA was prepared from a panel made up
of four Baka/a, three Bakb/b, and two Baka/b individuals, and the mRNA
encoding GPIIb was amplified using PCR and spotted onto nylon membranes.
ASO hybridization showed that the nucleotide base difference identified
above segregated with Bak phenotype in all nine individuals examined (P =
.002). The base pair substitution results in an amino acid polymorphism at
residue 843 of the mature heavy chain. The Baka form of GPIIb encodes an
isoleucine at this position, whereas the Bakb allele contains a serine.
Identification of the polymorphism associated with this clinically
important alloantigen system should permit new therapeutic and diagnostic
approaches for treating and managing patients with alloimmune
thrombocytopenic disorders.
Volume 75,
Issue 12,
pp. 2343-2348,
06/15/1990
Copyright © 1990 by The American Society of Hematology
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