IL-1-related cytokine responses of nonimmune skin cells subjected to CEES exposure with and without potential vesicant antagonists

Sulfur mustard provokes an acute inflammatory response in skin. To determine if keratinocytes regulate this response and whether three potential vesicant antagonists can counteract adverse changes, specimens of EpiDerm(TM) (MatTek Corp., Ashland, MA), a human skin model of differentiating keratinocytes, were exposed 2 h to humidified air with or without 2-chloroethyl ethyl sulfide (CEES, 1.72-1.73 mg/L/min) with or a without 10 mM niacinamide, a poly (ADP-ribose) polymerase (PARP) inhibitor, 25 mu M CGS9343B (calmodulin antagonist), or 8.4 mM leupeptin (cysteine protease inhibitor), After a 22-h incubation, levels of interleukin-1 alpha (IL-1 alpha), its receptor antagonist (IL-1Ra), soluble type II receptor (sIL-1RII and prostaglandin-E-2 (PGE(2)) were determined. Methylthiazole tetrazolium (MTT) viability tests and histological observations were also conducted. PGE(2) levels were abundant but unaffected by GEES regardless of antagonist presence. Total amounts (media plus lysate) of IL-1 alpha, IL-1Ra, and sIL-1RII were reduced with GEES irrespective of antagonist. GEES promoted the release of IL-1Ra, Exposure of EpiDerm to GEES in the presence of the vesicant antagonists did not improve viability or counteract histological damage. We conclude GEES depresses total IL-1 alpha and related cytokines, does not affect PGE2 release, and adverse changes associated with GEES-exposed EpiDerm are not ameliorated by these particular antagonists. Dramatically increased (5- to 10-fold) release of IL-1Ra may provide a useful marker for cytotoxicity, The high level of IL-1Ra and increased release with injury suggest a primary function in down-regulating IL-1 inflammatory responses in skin.