Comparison of cell size in sulfur mustard-induced death of keratinocytes and lymphocytes.
Sulfur mustard (HD) is a vesicant chemical warfare agent that directly alkylates cellular DNA and produces DNA strand breaks. To identify cellular models for in vitro screening of antivesicant compounds in DNA repair assays, we compared the mechanism of HD-induced cell death in cultured adult normal human epidermal keratinocytes (NHEK) and peripheral blood lymphocytes (PBL). One parameter that we used to distinguish apoptotic from necrotic cell death was the change in cell size due to HD. In the presence or absence of a poly(ADP-ribose) polymerase inhibitor (PARPI), cell preparations were exposed to various concentrations of HD (0.01-1.0 mM) and harvested at selected times after exposure (up to 24 h). Results from these experiments suggest that, with increasing HD concentration and time, NHEK will fragment irrespective of the presence or absence of PARPI, with cell fragmentation presumably preceded by necrosis. In the absence of PARPI, PBL size initially decreases and then remains constant over time. Previous DNA fragmentation studies indicate that both apoptosis and necrosis occur in HD-exposed PBL in a time-dependent manner. In the presence of PARPI, there is a HD concentration- and time-dependent decrease in PBL size that is characteristic of apoptosis. The shift in the mechanism of HD-induced PBL death from apoptosis followed by necrosis to exclusively apoptosis in the absence and presence of PARPI, respectively, is in agreement with previous findings on HD-induced changes in membrane integrity, energy levels and DNA fragmentation. Considering that NHEK fragment early after exposure to HD concentrations that produce vesication in human skin, PBL may be a more appropriate model for use in DNA repair assays.
Moser
J
J
Pharmacology Division, US Army Medical Research Institute of Chemical Defense, Aberdeen Proving Ground, MD 21010-5425, USA.
Meier
H L
HL
D017209:Apoptosis / Q000187:drug effects*; D001681:Biological Assay / Q000379:methods; D018929:Cell Culture Techniques; D048429:Cell Size; D004249:DNA Damage; D003879:Dermatologic Agents / Q000633:toxicity*; D004305:Dose-Response Relationship, Drug; D006801:Humans; D015603:Keratinocytes / Q000187:drug effects / Q000502:physiology; D008214:Lymphocytes / Q000187:drug effects / Q000502:physiology; D008954:Models, Biological; D009151:Mustard Gas / Q000633:toxicity*; D009336:Necrosis; D013997:Time Factors
D003879:Dermatologic Agents; D009151:Mustard Gas