Effect of HLA DR epitope de-immunization of Factor VIII in vitro and in vivo

Leonard Moise*, Chang Song, William D. Martin, Ryan Tassone, Anne S. De Groot, David W. Scott

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

66 Scopus citations

Abstract

T cell-dependent development of anti-Factor VIII (FVIII) antibodies that neutralize FVIII activity is a major obstacle to replacement therapy in hemophilia A. To create a less immunogenic therapeutic protein, recombinant FVIII can be modified to reduce HLA binding of epitopes based on predicted anchoring residues. Here, we used immunoinformatic tools to identify C2 domain HLA DR epitopes and predict site-specific mutations that reduce immunogenicity. Epitope peptides corresponding to original and modified sequences were validated in HLA binding assays and in immunizations of hemophilic E16 mice, DR3 and DR4 mice and DR3 × E16 mice. Consistent with immunoinformatic predictions, original epitopes are immunogenic. Immunization with selected modified sequences lowered immunogenicity for particular peptides and revealed residual immunogenicity of incompletely de-immunized modified peptides. The stepwise approach to reduce protein immunogenicity by epitope modification illustrated here is being used to design and produce a functional full-length modified FVIII for clinical use.

Original languageEnglish
Pages (from-to)320-331
Number of pages12
JournalClinical Immunology
Volume142
Issue number3
DOIs
StatePublished - Mar 2012
Externally publishedYes

Keywords

  • De-immunization
  • Epitope
  • Factor VIII
  • Inhibitors
  • T cell

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