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  • Assessment of the respiratory sensitization potential of proteins using an enhanced mouse intranasal test (MINT).

Assessment of the respiratory sensitization potential of proteins using an enhanced mouse intranasal test (MINT).

Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association (2013-06-12)
S M Krieger, D R Boverhof, M R Woolhiser, J A Hotchkiss
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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Proteinase, bacterial, Type XXIV, 7.0-14.0 units/mg solid, lyophilized powder
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Protease from Bacillus licheniformis, glycerol solution (50%)
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Protease from Bacillus licheniformis, Type VIII, lyophilized powder, 7-15 units/mg solid
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Protease from Bacillus licheniformis, lyophilized powder, for use in Total Dietary Fiber Assay, TDF-100A
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Protease from Bacillus licheniformis, ≥2.4 U/g