title: Aggregated recombinant human interferon Beta induces antibodies but no memory in immune-tolerant transgenic mice.
abstract: OBJECTIVE
  To study the influence of protein aggregation on the immunogenicity of recombinant human interferon beta (rhIFNbeta) in wild-type mice and transgenic, immune-tolerant mice, and to evaluate the induction of immunological memory.
  
  
  METHODS
  RhIFNbeta-1b and three rhIFNbeta-1a preparations with different aggregate levels were injected intraperitoneally in mice 15x during 3 weeks, and the mice were rechallenged with rhIFNbeta-1a. The formation of binding (BABs) and neutralizing antibodies (NABs) was monitored.
  
  
  RESULTS
  Bulk rhIFNbeta-1a contained large, mainly non-covalent aggregates and stressed rhIFNbeta-1a mainly covalent, homogeneous (ca. 100 nm) aggregates. Reformulated rhIFNbeta-1a was essentially aggregate-free. All products induced BABs and NABs in wild-type mice. Immunogenicity in the transgenic mice was product dependent. RhIFNbeta-1b showed the highest and reformulated rhIFNbeta-1a the lowest immunogenicity. In contrast with wild-type mice, transgenic mice did not show NABs, nor did they respond to the rechallenge.
  
  
  CONCLUSIONS
  The immunogenicity of the products in transgenic mice, unlike in wild-type mice, varied. In the transgenic mice, neither NABs nor immunological memory developed. The immunogenicity of rhIFNbeta in a model reflecting the human immune system depends on the presence and the characteristics of aggregates.
authors.affiliation: Department of Pharmaceutics Utrecht Institute for Pharmaceutical Sciences (UIPS), Utrecht University, Utrecht, The Netherlands. m.m.c.vanbeers@lacdr.leidenuniv.nl
authors.forename: Miranda M C
authors.initials: MM
authors.lastname: van Beers
authors.forename: Melody
authors.initials: M
authors.lastname: Sauerborn
authors.forename: Francesca
authors.initials: F
authors.lastname: Gilli
authors.forename: Vera
authors.initials: V
authors.lastname: Brinks
authors.forename: Huub
authors.initials: H
authors.lastname: Schellekens
authors.forename: Wim
authors.initials: W
authors.lastname: Jiskoot
mesh_terms: D000818:Animals; D057134:Antibodies, Neutralizing / Q000097:blood*; D001666:Binding Sites, Antibody; D015153:Blotting, Western; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007108:Immune Tolerance / Q000235:genetics; D007156:Immunologic Memory / Q000235:genetics; D007370:Interferon Type I / Q000009:adverse effects* / Q000235:genetics / Q000276:immunology*; D008027:Light; D051379:Mice; D008822:Mice, Transgenic; D011994:Recombinant Proteins; D012542:Scattering, Radiation; D013050:Spectrometry, Fluorescence
chemical_list: D057134:Antibodies, Neutralizing; D007370:Interferon Type I; D011994:Recombinant Proteins