Studies on the binding of an alloimmune and two murine monoclonal antibodies to the platelet glycoprotein IIb-IIIa complex receptor.
The platelet-binding characteristics of three different antibodies that completely block adenosine diphosphate (ADP)-induced platelet fibrinogen binding and react with glycoproteins IIb, IIIa, or both, were studied. Two of the antibodies are murine monoclonal antibodies (10E5 and 7E3), and the third is an alloimmune antibody produced by a patient with Glanzmann's thrombasthenia who has received multiple transfusions (E.S.). The two monoclonals differ in that 7E3, but not 10E5, binds to dog platelets as well as to human platelets. In mutual competition studies, 7E3 did not interfere with 10E5 binding, indicating that both antibodies could bind to the complex simultaneously. E.S. IgG produced only minor inhibition of 10E5 binding, but nearly complete inhibition of 7E3 binding, suggesting that its epitope lies closer to the 7E3 epitope than the 10E5 epitope. Ethylenediaminetetraacetic acid (EDTA) pretreatment of intact platelets at 22 degrees C and pH 7.75 did not affect 10E5 binding, but significantly inhibited 7E3 binding. Similar treatment of intact or solubilized platelets at high pH and temperature produced splitting of the glycoprotein IIb-IIIa (GPIIb-IIIa) complex and loss of both 10E5 and 7E3 binding. The 10E5 bound equally well to unactivated and activated platelets, and even rapid fixation of whole blood produced only a modest decrease in 10E5 binding. Preincubation of platelets with either E.S. IgG or 10E5 partially prevented EDTA from splitting the complex, and both monoclonal antibodies were able to bind to such complexes despite the EDTA treatment. These studies indicate that the binding of antibodies to several different epitopes on the GPIIb-IIIa complex can result in similar functional properties in blocking fibrinogen binding and platelet aggregation.(ABSTRACT TRUNCATED AT 250 WORDS)
B S
BS
Coller
E I
EI
Peerschke
U
U
Seligsohn
L E
LE
Scudder
A T
AT
Nurden
J P
JP
Rosa
D000244:Adenosine Diphosphate / Q000494:pharmacology; D000818:Animals; D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D001791:Blood Platelet Disorders / Q000097:blood*; D001792:Blood Platelets / Q000378:metabolism*; D002918:Chymotrypsin / Q000494:pharmacology; D015232:Dinoprostone; D004285:Dogs; D000939:Epitopes / Q000032:analysis; D011464:Epoprostenol / Q000494:pharmacology; D005340:Fibrinogen / Q000494:pharmacology; D006023:Glycoproteins / Q000378:metabolism*; D006801:Humans; D007126:Immunoglobulin Allotypes; D007493:Iraq / Q000208:ethnology; D007557:Israel; D008565:Membrane Proteins / Q000378:metabolism*; D010974:Platelet Aggregation / Q000187:drug effects; D010980:Platelet Membrane Glycoproteins; D011458:Prostaglandins E / Q000494:pharmacology; D011863:Radioimmunoassay; D011956:Receptors, Cell Surface / Q000187:drug effects / Q000378:metabolism*; D013915:Thrombasthenia / Q000097:blood*
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D000939:Epitopes; D006023:Glycoproteins; D007126:Immunoglobulin Allotypes; D008565:Membrane Proteins; D010980:Platelet Membrane Glycoproteins; D011458:Prostaglandins E; D011956:Receptors, Cell Surface; D000244:Adenosine Diphosphate; D005340:Fibrinogen; D011464:Epoprostenol; D002918:Chymotrypsin; D015232:Dinoprostone