Procedure for assessing myeloperoxidase and inflammatory mediator responses in hairless mouse skin

A preparation procedure for making multiple inflammatory biomarker measurem ents from the same skin tissue was assessed. The backs of euthymic hairless mice were exposed to sulfur mustard (HD) vapor for 6 min. Animals were eut hanized 24 h following exposure, dorsal skin tissue was excised and 12-mm, full-thickness biopsy punches of the exposed skin sites were taken. Specime ns were snap-frozen, crushed to a powder using a hiopulverizer unit, solubi lized in buffer and centrifuged. Supernatant was assayed for pro-inflammato ry cytokines and the acute-phase reactive protein, serum amyloid P (SAP). M yeloperoxidase (MPX), which is indicative of neutrophil infiltration into t he skin, was associated with the pellet fraction. Results indicate an eleva tion of interleukin-6, SAP and MPX in mouse skin tissue specimens 24 h foll owing HD vapor exposure. The tissue preparation procedure allows the use of a single skin specimen to make multiple inflammatory endpoint measurements requiring different preparation processes, and it will be used in subseque nt studies to characterize further the inflammatory nature of HD-exposed sk in tissue. Copyright (C) 2000 John Wiley & Sons, Ltd.