title: Assessment of the respiratory sensitization potential of proteins using an enhanced mouse intranasal test (MINT).
abstract: There remains a need for a simple and predictive animal model to identify potential respiratory sensitizers. The mouse intranasal test (MINT) was developed to assess the relative allergic potential of detergent enzymes, however, the experimental endpoints were limited to evaluation of antibody levels. The present study was designed to evaluate additional endpoints (serum and allergic antibody levels, pulmonary inflammation and airway hyperresponsiveness (AHR)) to determine their value in improving the predictive accuracy of the MINT. BDF1 mice were intranasally instilled on days 1, 3, 10, 17 and 24 with subtilisin, ovalbumin, betalactoglobulin, mouse serum albumin or keyhole limpet hemocyanin; challenged with aerosolized methacholine or the sensitizing protein on day 29 to assess AHR, and sacrificed on day 29 or 30. Under the conditions of this study, evaluation of AHR did not improve the predictive power of this experimental model. Allergic antibody responses and IgG isotype characterization proved to be the most sensitive and reliable indicators of the protein allergenic potential with BAL responses providing additional insight. These data highlight that the evaluation of the respiratory sensitization potential of proteins can be best informed when multiple parameters are evaluated and that further improvements and refinements of the assay are necessary.
authors.affiliation: Toxicology & Environmental Research and Consulting, The Dow Chemical Company, Midland, MI 48674, United States.
authors.forename: S M
authors.initials: SM
authors.lastname: Krieger
authors.forename: D R
authors.initials: DR
authors.lastname: Boverhof
authors.forename: M R
authors.initials: MR
authors.lastname: Woolhiser
authors.forename: J A
authors.initials: JA
authors.lastname: Hotchkiss
mesh_terms: D000281:Administration, Intranasal; D000336:Aerosols; D000485:Allergens / Q000008:administration & dosage / Q000009:adverse effects*; D000818:Animals; D000906:Antibodies / Q000032:analysis; D001992:Bronchoalveolar Lavage Fluid / Q000276:immunology; D004044:Dietary Proteins / Q000008:administration & dosage / Q000009:adverse effects; D004306:Dose-Response Relationship, Immunologic; D005260:Female; D007074:Immunoglobulin G / Q000032:analysis; D007782:Lactoglobulins / Q000008:administration & dosage / Q000009:adverse effects*; D051379:Mice; D008815:Mice, Inbred Strains; D023421:Models, Animal; D020556:Neutrophil Infiltration / Q000187:drug effects; D010047:Ovalbumin / Q000008:administration & dosage / Q000009:adverse effects*; D011014:Pneumonia / Q000209:etiology; D015203:Reproducibility of Results; D012130:Respiratory Hypersensitivity / Q000139:chemically induced* / Q000276:immunology / Q000473:pathology / Q000503:physiopathology; D020545:Respiratory Mucosa / Q000187:drug effects* / Q000276:immunology / Q000473:pathology; D020860:Subtilisin / Q000008:administration & dosage / Q000009:adverse effects*
chemical_list: D000336:Aerosols; D000485:Allergens; D000906:Antibodies; D004044:Dietary Proteins; D007074:Immunoglobulin G; D007782:Lactoglobulins; D010047:Ovalbumin; D020860:Subtilisin
keywords: AB/PAS; AHR; ANOVA; Alcian Blue/Periodic Acid Schiff sequence; Asthma; BAL; BLG; ELISAA; H&E; HRP; IACUC; Institutional Animal Care and Use Committee; KLH; MCh; MHC; MINT; MSA; MV; Mice; OVA; PCA; Penh; Protein allergy; Respiratory sensitization; SUB; TV; WBP; Whole-body plethysmography; airway hyperresponsiveness; analysis of variance; beta-lactoglobulin; bronchoalveolar lavage; enhanced pause; enzyme-linked immunosorbent assay; hematoxylin and eosin; horseradish peroxidase; keyhole limpet hemocyanin; major histocompatibility complex; methacholine; minute volume; mouse intranasal test; mouse serum albumin; ovalbumin; passive cutaneous anaphylaxis; subtilisin; tidal volume; whole-body plethysmography