Response of normal human keratinocytes to sulfur mustard: Cytokine release

Cytokines play a major role in both acute and chronic inflammatory processe s, including those produced by sulfur mustard (2,2'-dichlorodiethyl sulfide , HD). This study describes responses of normal human epidermal keratinocyt es (NHEK) to HD, defined by interleukin-1 beta (IL-1 beta), IL-6, IL-8 and tumor necrosis factor alpha (TNF-ol) release. Commercially available enzyme -linked immunosorbent assay (ELISA) kits were used to measure the cytokine release in NHEK during exposure to 100 and 300 muM of HD. Exposure to 100 m uM HD increased the release of cytokines, The amounts of IL-8 and TNF-alpha present in cell suspensions increased up to 59-fold and 4-fold, respective ly, above control levels when NHEK were exposed to 300 muM HD. Exposure of NHEK to 300 muM HD had a highly variable effect on the release of IL-1 beta , where sometimes the secretion of IL-1 beta increased above baseline level and at other times it decreased in cell suspensions. Supernatants were col lected from cell culture flasks 24 h after exposure of 100 and 300 muM HD a nd significantly increased levels of 1L-6 were observed. Interleukin-6 was released in a concentration-dependent manner, 3.6-fold up to 8.4-fold, resp ectively, in supernatant, These pro-inflammatory mediators IL-I beta, IL-8, TNP-alpha and IL-6 may play an important role in HD injury. The present fi ndings suggest that the cytokine changes detected could be used as potentia l biomarkers of cutaneous vesicant injury.