Aggregated recombinant human interferon Beta induces antibodies but no memory in immune-tolerant transgenic mice.
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.
van Beers
Miranda M C
MM
Department of Pharmaceutics Utrecht Institute for Pharmaceutical Sciences (UIPS), Utrecht University, Utrecht, The Netherlands. m.m.c.vanbeers@lacdr.leidenuniv.nl
Sauerborn
Melody
M
Gilli
Francesca
F
Brinks
Vera
V
Schellekens
Huub
H
Jiskoot
Wim
W
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
D057134:Antibodies, Neutralizing; D007370:Interferon Type I; D011994:Recombinant Proteins