Localization of human factor FVIII inhibitor epitopes to two polypeptide fragments.
Epitopes for 22 alloantibodies that inhibit factor VIII procoagulant protein (FVIII) from multitransfused individuals with severe hemophilia A and three autoantibodies from nonhemophilic individuals appeared to be restricted to two specific regions of the FVIII molecule. Immunoblotting of purified FVIII and purified thrombin-degraded FVIII, followed by reaction with inhibitor plasma samples, monoclonal anti-human IgG3 and IgG4 antibodies, and radiolabeled affinity-purified rabbit anti-mouse IgG, revealed that inhibitor epitopes could be localized to the Mr 72,000 and Mr 44,000 thrombin fragments of FVIII. These two chains are located at the carboxyl terminus and near the amino terminus of the FVIII molecule, respectively. The pattern of reactivity of the inhibitor alloantibodies could be divided into three types: 10 reacted with the Mr 72,000 chain, 3 reacted with the Mr 44,000 chain, and 9 reacted with both of these chains. Among the 3 inhibitor autoantibodies, 1 of each type was found. Ten normal plasmas, as well as 14 plasmas from multitransfused individuals with severe hemophilia A and no inhibitor, were not reactive with the FVIII immunoblots. However, one multitransfused individual with severe hemophilia A and no detectable inhibitor revealed the presence of an antibody reactive with the middle section of the FVIII molecule. The existence of FVIII inhibitor epitopes on both the Mr 72,000 and Mr 44,000 chains raises the possibility that these epitopes might be further restricted to regions of homology between the two chains. These data suggest the possibility of designing inhibitor blocking polypeptides for use as therapeutic agents.
C A
CA
Fulcher
S
S
de Graaf Mahoney
J R
JR
Roberts
C K
CK
Kasper
T S
TS
Zimmerman
D001323:Autoantibodies / Q000276:immunology; D000939:Epitopes / Q000032:analysis*; D005169:Factor VIII / Q000037:antagonists & inhibitors* / Q000276:immunology / Q000302:isolation & purification; D006467:Hemophilia A / Q000276:immunology*; D006801:Humans; D007518:Isoantibodies / Q000276:immunology*; D008970:Molecular Weight; D010446:Peptide Fragments / Q000276:immunology*
D001323:Autoantibodies; D000939:Epitopes; D007518:Isoantibodies; D010446:Peptide Fragments; D005169:Factor VIII