Specific antibody responses to subtilisin Carlsberg (Alcalase) in mice: Development of an intranasal exposure model

Abstract

An intranasal (i.n.) dosing model was developed in mice as a potential alternative to more difficult, time-consuming, and costly guinea pig intratracheal (GPIT) or mouse intratracheal models for assessment of the respiratory immunogenicity of detergent en-zymes. Using a benchmark enzyme, Alcalase (protease subtilisin Carlsberg), studies were conducted to standardize the model In terms of mouse strain, dosing and serum harvest regimen, and the primary immunoglobulin endpoint to use. The primary assay endpoint selected was the enzyme-specific IgGl titer determined by an Alcalase-speciflc ELISA. This is not the primary allergenic antibody in mice (IgE is); however, IgGl is coregulated with IgE via the IL-4/TH2 pathway and may have a role in mediating allergic-type responses. BDF1 mice (C57B1/6 x DBA/2) were se-lected as representative of high responder strains, with high re-sponse associated with the H-2 " (C57B176) parent. The dosing regi