Development and utilization of an ex vivo bromodeoxyuridine local lymph node assay protocol for assessing potential chemical sensitizers

摘要: The murine local lymph node assay (LLNA) is widely used to identify chemicals that may cause allergic contact dermatitis. Exposure a dermal sensitizer results in proliferation of T cells, which has traditionally been measured by vivo incorporation [3H]methyl thymidine. A more recent non-isotopic variation the utilizes bromodeoxyuridine (BrdU) vivo. To further improve utility this assay, we developed an ex BrdU labeling procedure eliminating need for injections. correctly identified strong (i.e., trimellitic anhydride) as well weak/moderate sensitizers eugenol, cinnamaldehyde and hexylcinnaminic aldehyde). As anticipated, neither non-sensitizers isopropanol lactic acid nor false negative chemical nickel II sulfate hexahydrate induced positive threshold response assay. are close agreement with those LLNA:BrdU-enzyme-linked immunosorbent procedure. We also LLNA evaluate ammonium hexachloroplatinate, tetrachloroplatinate cis-diamminedichloroplatinum(II) them based on calculation EC2 values. conclude method offers predictive capacity comparable previously established protocols while animal injections use radioisotope. Published 2014. This article U.S. Government work public domain USA.