Alloantibody from a patient with severe von Willebrand disease inhibits von Willebrand factor-FVIII interaction.
The aim of this study was to analyze the ability of an alloantibody from a patient with severe von Willebrand disease (vWD) to interfere with the vWF domain for FVIII, to inhibit factor VIII (FVIII), and to compare it with a rabbit polyclonal antibody. The vWF domain for binding to FVIII was assayed by a method previously described but using recombinant FVIII (r-FVIII, Kogenate), which contains no vWF, instead of Hemofil M (HM). Rabbit or human antibodies towards FVIII (FVIII-Ab) were analyzed using microtiter wells with immobilized r-FVIII through a monoclonal anti-FVIII antibody and an ELISA method. IgG from plasma of a patient with hemophilia A and FVIII inhibitor was used as a positive control. Normal human and rabbit IgGs were included as negative controls. Human vWD alloantibody IgG and the rabbit anti-vWF antibody IgG reacted with immobilized normal vWF, inhibiting its binding to r-FVIII in a dose-dependent manner, which suggests that it is specific. Normal human IgG fraction, as well as nonspecific rabbit IgG, did not interfere with this binding at all. The monoclonal antibody used in this assay to immobilize vWF did not alter this interaction at all. Human vWD alloantibody IgG and the rabbit antibody against vWF showed a partial inhibitory activity to plasma FVIII as well as r-FVIII. The inhibition reached a plateau with residual FVIII activity. FVIII-Ab were not detected in human alloantibody or in rabbit antibody preparations. In contrast, hemophiliac FVIII inhibitor showed FVIII-AB. This human vWD alloantibody behaves like polyclonal heterologous antibodies, and their inhibition of FVIII seems to be nonspecific due to a steric hindrance mechanism provided that both have no FVIII antibodies.
Batlle
J
J
Department of Hematology, Hospital Juan Canalejo-Teresa Herrera, La Coruña, Spain.
Lourés
E
E
Vila
P
P
Hernández
M C
MC
Méndez
J A
JA
Torea
J
J
Rendal
E
E
Couselo
M J
MJ
Filgueira
A
A
López Fernández
M F
MF
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000937:Antigen-Antibody Reactions; D005169:Factor VIII / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D006720:Homozygote; D006801:Humans; D007518:Isoantibodies / Q000494:pharmacology*; D011485:Protein Binding / Q000187:drug effects; D011817:Rabbits; D014842:von Willebrand Diseases / Q000097:blood / Q000276:immunology*; D014841:von Willebrand Factor / Q000037:antagonists & inhibitors*
D000911:Antibodies, Monoclonal; D007518:Isoantibodies; D014841:von Willebrand Factor; D005169:Factor VIII