Phenotypes of Allo- and Autoimmune Antibody Responses to FVIII Characterized by Surface Plasmon Resonance

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  • Lewis, Kenneth B.
  • Hughes, Richard J.
  • Epstein, Melinda S.
  • Josephson, Neil C.
  • Kempton, Christine L.
  • Kessler, Craig M.
  • Key, Nigel S.
  • Howard, Tom E.
  • Kruse-Jarres, Rebecca
  • Lusher, Jeanne M.
  • Walsh, Christopher E.
  • Watts, Raymond G.
  • Ettinger, Ruth A.
  • Pratt, Kathleen P.
Abstract
  • Evidence of antibody isotype/subtype switching may provide prognostic value regarding the state of immune responses to therapeutic proteins, e.g. anti-factor VIII (FVIII) antibodies that develop in many hemophilia A patients, clinically termed “inhibitors”. A sensitive, high- information-content surface plasmon resonance (SPR) assay has been developed to quantify IgG subtype distributions and the domain specificity of anti-drug antibodies. Plasma samples from 22 subjects with an allo- or auto-immune reaction to FVIII were analyzed. Pre-analytical treatment protocols were developed to minimize non-specific binding and specific matrix interference due to von Willebrand factor-FVIII interactions. The dynamic range for IgG quantification was 0.2–5 µg/ml (∼1–33 nM), allowing characterization of inhibitor-positive samples. Subtype-specific monoclonal antibodies were used to quantify the IgG subtype distribution of FVIII-specific antibodies. Most samples obtained from multiply-infused inhibitor subjects contained IgG4 antibodies. Several distinct phenotypes were assigned based on the IgG subtype distribution: IgG1, IgG4, IgG1 & IgG4, and IgG1, IgG2 & IgG4. An IgG1-only response was found in mild/moderate HA subjects during early FVIII infusions, and analysis of serial samples followed antibody class switching as several subjects’ immune responses developed. Competition studies utilizing a recombinant FVIII-C2 domain indicated 40–80% of FVIII-specific antibodies in most samples were directed against this domain.
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  • Article
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  • In Copyright
Journal title
  • PloS One
Journal volume
  • 8
Journal issue
  • 5
Page start
  • e61120
Language
  • English
ISSN
  • 1932-6203

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