title: The missense mutation Arg593 --> Cys is related to antibody formation in a patient with mild hemophilia A.
abstract: The development of inhibitory antibodies to factor VIII in patients affected by a mild form of hemophilia A (factor VIII > 0.05 IU/mL) is considered a rare event. In this study, we evaluated the relationship between genotype and anti-factor VIII antibody formation in a patient with mild hemophilia A. Mutation analysis showed that a missense mutation in the factor VIII gene leading to replacement of Arg593 by Cys in the A2 domain of factor VIII was associated with hemophilia A in this patient. The anti-factor VIII antibodies present in the patient's plasma were characterized using metabolically labeled factor VIII fragments expressed in insect cells. The anti-factor VIII antibodies, composed of subclasses IgG2 and IgG4, reacted with both the fragment corresponding to the factor VIII heavy chain and the A2 domain. The Arg593 --> Cys substitution was introduced into the cDNA encoding the A2 domain of factor VIII and the resulting construct was expressed in insect cells. Strikingly, the metabolically labeled A2 domain carrying the Arg593 --> Cys mutation was not recognized by the anti-factor VIII antibodies present in the plasma of the patient. These data indicate that the anti-factor VIII antibodies are exclusively directed against exogenous factor VIII. This strongly suggests that the Arg593 --> Cys substitution results in recognition of wild-type factor VIII as nonself and is thereby related to the formation of anti-factor VIII antibodies after factor VIII replacement therapy in this particular patient.
authors.affiliation: Department of Blood Coagulation, Central Laboratory of the Netherlands Red Cross Blood Transfusion Service, Amsterdam.
authors.forename: K
authors.initials: K
authors.lastname: Fijnvandraat
authors.forename: E A
authors.initials: EA
authors.lastname: Turenhout
authors.forename: E N
authors.initials: EN
authors.lastname: van den Brink
authors.forename: J W
authors.initials: JW
authors.lastname: ten Cate
authors.forename: J A
authors.initials: JA
authors.lastname: van Mourik
authors.forename: M
authors.initials: M
authors.lastname: Peters
authors.forename: J
authors.initials: J
authors.lastname: Voorberg
mesh_terms: D000328:Adult; D001324:Autoantigens / Q000737:chemistry / Q000235:genetics; D004200:Diseases in Twins; D000939:Epitopes / Q000737:chemistry / Q000235:genetics; D005169:Factor VIII / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D006467:Hemophilia A / Q000235:genetics*; D006801:Humans; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000276:immunology; D007518:Isoantibodies / Q000096:biosynthesis*; D007519:Isoantigens / Q000737:chemistry; D008297:Male; D017354:Point Mutation; D014430:Twins, Monozygotic
chemical_list: D001324:Autoantigens; D000939:Epitopes; D007074:Immunoglobulin G; D007518:Isoantibodies; D007519:Isoantigens; D005169:Factor VIII