Fine specificity of drug-dependent antibodies reactive with a restricted domain of platelet GPIIIA.
Drug-induced immune thrombocytopenia is caused by drug-dependent antibodies (DDAbs) that bind tightly to platelet glycoproteins only when drug is present. How drugs mediate this interaction is not yet resolved. Several studies indicate that sites recognized by DDAbs tend to cluster in specific structural domains, suggesting they may recognize a limited number of distinct epitopes. To address this issue, we characterized the binding sites for 16 quinine-dependent antibodies thought on the basis of preliminary studies to be possibly specific for a single epitope on glycoprotein IIIa (GPIIIa). Fourteen of the antibodies reacted with a 29-kDa GPIIIa fragment comprising only the GPIIIa hybrid and plextrin-semaphorin-integrin homology domains. However, studies with mutant GPIIIa and the blocking monoclonal antibody AP3 showed that the 14 DDAbs recognize at least 6 and possibly more distinct, but overlapping, structures involving GPIIIa residues 50 to 66. The findings suggest that even antibodies specific for restricted domains on a target glycoprotein may each have a slightly different fine specificity; ie, "unique" epitopes recognized by DDAbs may be rare or nonexistent. The observations are consistent with a recently proposed model in which drug reacts noncovalently with both target protein and antibody to promote binding of an otherwise nonreactive immunoglobulin.
Peterson
Julie A
JA
Blood Research Institute, BloodCenter of Wisconsin, Milwaukee, WI 53201-2178, USA. jukie.peterson@bcw.edu
Nelson
Tamara N
TN
Kanack
Adam J
AJ
Aster
Richard H
RH
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology*; D001665:Binding Sites; D002460:Cell Line; D006801:Humans; D039661:Integrin beta3 / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D008969:Molecular Sequence Data; D004364:Pharmaceutical Preparations; D012680:Sensitivity and Specificity; D016415:Sequence Alignment
D000906:Antibodies; D039661:Integrin beta3; D004364:Pharmaceutical Preparations