title: Development of a transgenic mouse model to study the immunogenicity of recombinant human insulin.
abstract: Mouse models are commonly used to assess the immunogenicity of therapeutic proteins and to investigate the immunological processes leading to antidrug antibodies. The aim of this work was to develop a transgenic (TG) Balb/c mouse model for evaluating the immunogenicity of recombinant human insulin (insulin) formulations. Validation of the model was performed by measuring the antibody response against plain and particulate insulin in TG and nontransgenic (NTG) mice. Intraperitoneal administration of insulin (20 μg/dose, 12 doses over a period of 4 weeks) did not break the immune tolerance of the TG mice, whereas it did elicit antibodies in NTG mice. The immune tolerance of TG mice could be circumvented, albeit at low titers, by administering insulin covalently bound to 50-nm polystyrene nanoparticles. The TG mouse model was employed to compare the immunogenicity of oxidized aggregated insulin, oxidized nonaggregated insulin, and three commercially available formulations of insulin variants (i.e., Levemir®, Insulatard®, and Actrapid®). Oxidized insulin, aggregated or nonaggregated, was moderately immunogenic in TG mice (50% and 33% responders, respectively), whereas the immunogenicity of the commercial formulations was low. This model can be used to compare the immunogenicity of insulin formulations and to study immune mechanisms of antibody formation against insulin.
authors.affiliation: Division of Drug Delivery Technology, Leiden Academic Centre for Drug Research (LACDR), Leiden University, Leiden, 2300, RA, The Netherlands.
authors.forename: Riccardo
authors.initials: R
authors.lastname: Torosantucci
authors.forename: Vera
authors.initials: V
authors.lastname: Brinks
authors.forename: Grzegorz
authors.initials: G
authors.lastname: Kijanka
authors.forename: Liem Andhyk
authors.initials: LA
authors.lastname: Halim
authors.forename: Melody
authors.initials: M
authors.lastname: Sauerborn
authors.forename: Huub
authors.initials: H
authors.lastname: Schellekens
authors.forename: Wim
authors.initials: W
authors.lastname: Jiskoot
mesh_terms: D000818:Animals; D000917:Antibody Formation / Q000276:immunology*; D002626:Chemistry, Pharmaceutical / Q000379:methods; D006801:Humans; D007108:Immune Tolerance / Q000276:immunology; D007328:Insulin / Q000737:chemistry / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008822:Mice, Transgenic / Q000276:immunology*; D053758:Nanoparticles / Q000737:chemistry; D010316:Particle Size; D011137:Polystyrenes / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology*
chemical_list: D007328:Insulin; D011137:Polystyrenes; D011994:Recombinant Proteins
keywords: immune response; nanoparticles; oxidation; physicochemical properties; protein aggregation; protein formulation