Development of an In Vitro Assay of Mustard-Induced Proteolysis Using Human Cells in Culture

摘要

This report describes the development of in vitro assays of mustard- induced proteolysis using human cells in culture. Development of rapid, cost- effective, and relevant assays is important for the screening of the many potential drugs that are available from pharmaceutical companies or developed under contract. Efficacy of these prospective antidotes is measured using salient biological endpoints. Proteolysis appears to be involved in the pathogenesis of the sulfur mustard (HD) lesion because of the formation of blisters at the epidermal-dermal junction. A serine protease seems to be responsible for this formation and is present in some in vitro human models used at this facility. Protease release and its inhibition may be measured by chromogenic assays using Chromozym TH in selected donor peripheral blood lymphocytes (PBL). In studies with human epidermal keratinocytes (HEK), chromogenic protease activity was demonstrated between 100 and 300 1%M HD with Chromozym U and Chromozym t-PA. However, predictability of protease expression in HEK has not yet been achieved.